bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2021–12–26
75 papers selected by
Viktor Korolchuk, Newcastle University
  1. New regulators of PRKN-independent mitophagy.
  2. Ion Channels and Pumps in Autophagy: A Reciprocal Relationship.
  3. Autophagy-Lysosomal Pathway as Potential Therapeutic Target in Parkinson's Disease.
  4. Mitophagy in Yeast: Molecular Mechanism and Regulation.
  5. Endophilin-B regulates autophagy during synapse development and neurodegeneration.
  6. Autophagy modulates cell fate decisions during lineage commitment.
  7. The Role of Chaperone-Mediated Autophagy in Hepatitis C Virus-Induced Pathogenesis.
  8. Mitophagy in neurological disorders.
  9. Mitophagy in Yeast: Decades of Research.
  10. p38-TFEB pathways promote microglia activation through inhibiting CMA-mediated NLRP3 degradation in Parkinson's disease.
  11. Emerging role of mitophagy in myoblast differentiation and skeletal muscle remodeling.
  12. Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy.
  13. Protein Kinase CK2 Is Upregulated by Calorie Restriction and Induces Autophagy.
  14. BNIP3L/NIX-mediated mitophagy: molecular mechanisms and implications for human disease.
  15. Mechanisms of FA-Phagy, a New Form of Selective Autophagy/Organellophagy.
  16. Parkin beyond Parkinson's Disease-A Functional Meaning of Parkin Downregulation in TDP-43 Proteinopathies.
  17. Phospholipase Dε Interacts with Autophagy-Related-Protein 8 and Promotes Autophagy in Arabidopsis Response to Nitrogen Deficiency.
  18. Fuel for emotion: how mitophagy in the BLA can mediate increased anxiety in stressful social situations.
  19. TFEB-dependent autophagy is involved in scavenger receptor OLR1/LOX-1-mediated tumor progression.
  20. Molecular Signaling to Preserve Mitochondrial Integrity against Ischemic Stress in the Heart: Rescue or Remove Mitochondria in Danger.
  21. Amyloid-like aggregating proteins cause lysosomal defects in neurons via gain-of-function toxicity.
  22. An ALS-associated variant of the autophagy receptor SQSTM1/p62 reprograms binding selectivity towards the autophagy-related hATG8 proteins.
  23. Formimidoyltransferase cyclodeaminase prevents the starvation-induced liver hepatomegaly and dysfunction through downregulating mTORC1.
  24. Emerging Role of Autophagy in the Development and Progression of Oral Squamous Cell Carcinoma.
  25. HEXA-018, a Novel Inducer of Autophagy, Rescues TDP-43 Toxicity in Neuronal Cells.
  26. Protein Quality Control at the Mitochondrial Surface.
  27. Impairment of lysosomal function by Clostridioides difficile TcdB.
  28. Activation mechanism of PINK1.
  29. Distinct subcellular autophagy impairments in induced neurons from Huntington's disease patients.
  30. Protein Aggregation in the ER: Calm behind the Storm.
  31. RalA, PLD and mTORC1 Are Required for Kinase-Independent Pathways in DGKβ-Induced Neurite Outgrowth.
  32. CRISPR screens for lipid regulators reveal a role for ER-bound SNX13 in lysosomal cholesterol export.
  33. Activation of Downstream mTORC1 Target Ribosomal Protein S6 Kinase (S6K) Can Be Found in a Subgroup of Dutch Patients with Granulomatous Pulmonary Disease.
  34. The two-domain architecture of LAMP2A regulates its interaction with Hsc70.
  35. Identification of Novel Therapeutic Targets for Polyglutamine Diseases That Target Mitochondrial Fragmentation.
  36. Cell Survival and Cell Death at the Intersection of Autophagy and Apoptosis: Implications for Current and Future Cancer Therapeutics.
  37. Apigenin Induces Autophagy and Cell Death by Targeting EZH2 under Hypoxia Conditions in Gastric Cancer Cells.
  38. Targeting FAM134B-mediated reticulophagy activates sorafenib-induced ferroptosis in hepatocellular carcinoma.
  39. The Bumpy Road towards mTOR Inhibition in Glioblastoma: Quo Vadis?
  40. How the Innate Immune DNA Sensing cGAS-STING Pathway Is Involved in Autophagy.
  41. The Autophagic Route of E-Cadherin and Cell Adhesion Molecules in Cancer Progression.
  42. Aging of Podospora anserina Leads to Alterations of OXPHOS and the Induction of Non-Mitochondrial Salvage Pathways.
  43. Ellagic Acid Prevents α-Synuclein Aggregation and Protects SH-SY5Y Cells from Aggregated α-Synuclein-Induced Toxicity via Suppression of Apoptosis and Activation of Autophagy.
  44. ROS-Induced mtDNA Release: The Emerging Messenger for Communication between Neurons and Innate Immune Cells during Neurodegenerative Disorder Progression.
  45. MAP4K4 mediates the SOX6-induced autophagy and reduces the chemosensitivity of cervical cancer.
  46. The MAP Kinase Phosphatase MKP-1 Modulates Neurogenesis via Effects on BNIP3 and Autophagy.
  47. Autophagy Promotes Hepatic Cystogenesis in Polycystic Liver Disease via Depletion of Cholangiocyte Ciliogenic Proteins.
  48. Sidt2 is a key protein in the autophagy-lysosomal degradation pathway and is essential for the maintenance of kidney structure and filtration function.
  49. Implication of type 4 NADPH oxidase (NOX4) in tauopathy.
  50. Impaired Retromer Function in Niemann-Pick Type C Disease Is Dependent on Intracellular Cholesterol Accumulation.
  51. Autophagy Is Involved in the Viability of Overexpressing Thioredoxin o1 Tobacco BY-2 Cells under Oxidative Conditions.
  52. Interdependent Regulation of Polycystin Expression Influences Starvation-Induced Autophagy and Cell Death.
  53. Disruption of the TCA cycle reveals an ATF4-dependent integration of redox and amino acid metabolism.
  54. Polygala saponins inhibit NLRP3 inflammasome-mediated neuroinflammation via SHP2-Mediated mitophagy.
  55. mTOR Inhibition via Rapamycin Treatment Partially Reverts the Deficit in Energy Metabolism Caused by FH Loss in RPE Cells.
  56. Advances in Proteasome Enhancement by Small Molecules.
  57. Targeting PLD2 in adipocytes augments adaptive thermogenesis by improving mitochondrial quality and quantity in mice.
  58. Indoxyl Sulfate Contributes to mTORC1-Induced Renal Fibrosis via The OAT/NADPH Oxidase/ROS Pathway.
  59. Biomimetic nanoparticles blocking autophagy for enhanced chemotherapy and metastasis inhibition via reversing focal adhesion disassembly.
  60. Amino acid transporters as modulators of glucose homeostasis.
  61. An Inhibitor of Grp94 Inhibits OxLDL-Induced Autophagy and Apoptosis in VECs and Stabilized Atherosclerotic Plaques.
  62. Sequestosome 1 Is Part of the Interaction Network of VAPB.
  63. Metformin Improves Stemness of Human Adipose-Derived Stem Cells by Downmodulation of Mechanistic Target of Rapamycin (mTOR) and Extracellular Signal-Regulated Kinase (ERK) Signaling.
  64. Therapeutic Approaches in Lysosomal Storage Diseases.
  65. Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators.
  66. Camphorquinone Promotes the Antisenescence Effect via Activating AMPK/SIRT1 in Stem Cells and D-Galactose-Induced Aging Mice.
  67. HDAC6 Inhibition Extinguishes Autophagy in Cancer: Recent Insights.
  68. Long non-coding RNA TUG1 promotes the osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the AMPK/mTOR/autophagy pathway.
  69. Crosstalk Between Autophagy and the cGAS-STING Signaling Pathway in Type I Interferon Production.
  70. SQSTM1-mediated clearance of cytoplasmic mutant TARDBP/TDP-43 in the monkey brain.
  71. Resolvin D1 Suppresses H2O2-Induced Senescence in Fibroblasts by Inducing Autophagy through the miR-1299/ARG2/ARL1 Axis.
  72. The kinase complex mTORC2 promotes the longevity of virus-specific memory CD4+ T cells by preventing ferroptosis.
  73. Early lysosome defects precede neurodegeneration with amyloid-β and tau aggregation in NHE6-null rat brain.
  74. The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma.
  75. The role of DRP1- PINK1-Parkin-mediated mitophagy in early cadmium-induced liver damage.
  1. Autophagy. 2021 Dec 19. 1-3
    New regulators of PRKN-independent mitophagy.
    Yuchen Lei, Daniel J Klionsky.
      Mitophagy, a type of selective autophagy targeting damaged or superfluous mitochondria, is critical to maintain cell homeostasis. Besides the well-characterized PRKN-dependent mitophagy, PRKN-independent mitophagy also plays significant physiological roles. In a recent study, researchers from Anne Simonsen's lab discovered two lipid binding kinases, GAK and PRKCD, as positive regulators of PRKN-independent mitophagy. The researchers further investigated how these two proteins regulate mitophagy and demonstrated their roles in vivo. Focusing on the less known PRKN-independent mitophagy regulators, these findings shed light on understanding the mechanism of mitophagy and its relation to diseases.
    Keywords:  Autophagy; GAK; PRKCD; PRKN; lipid-binding kinases
    DOI:  https://doi.org/10.1080/15548627.2021.2012867
  2. Cells. 2021 Dec 14. pii: 3537. [Epub ahead of print]10(12):
    Ion Channels and Pumps in Autophagy: A Reciprocal Relationship.
    Hussein Abuammar, Arindam Bhattacharjee, Zsófia Simon-Vecsei, András Blastyák, Gábor Csordás, Tibor Páli, Gábor Juhász.
      Autophagy, the process of cellular self-degradation, is intrinsically tied to the degradative function of the lysosome. Several diseases have been linked to lysosomal degradative defects, including rare lysosomal storage disorders and neurodegenerative diseases. Ion channels and pumps play a major regulatory role in autophagy. Importantly, calcium signaling produced by TRPML1 (transient receptor potential cation channel, mucolipin subfamily) has been shown to regulate autophagic progression through biogenesis of autophagic-lysosomal organelles, activation of mTORC1 (mechanistic target of rapamycin complex 1) and degradation of autophagic cargo. ER calcium channels such as IP3Rs supply calcium for the lysosome, and lysosomal function is severely disrupted in the absence of lysosomal calcium replenishment by the ER. TRPML1 function is also regulated by LC3 (microtubule-associated protein light chain 3) and mTORC1, two critical components of the autophagic network. Here we provide an overview of the current knowledge about ion channels and pumps-including lysosomal V-ATPase (vacuolar proton-ATPase), which is required for acidification and hence proper enzymatic activity of lysosomal hydrolases-in the regulation of autophagy, and discuss how functional impairment of some of these leads to diseases.
    Keywords:  TRPML; V-ATPase; autophagy; calcium; ion channels
    DOI:  https://doi.org/10.3390/cells10123537
  3. Cells. 2021 Dec 15. pii: 3547. [Epub ahead of print]10(12):
    Autophagy-Lysosomal Pathway as Potential Therapeutic Target in Parkinson's Disease.
    Srinivasa Reddy Bonam, Christine Tranchant, Sylviane Muller.
      Cellular quality control systems have gained much attention in recent decades. Among these, autophagy is a natural self-preservation mechanism that continuously eliminates toxic cellular components and acts as an anti-ageing process. It is vital for cell survival and to preserve homeostasis. Several cell-type-dependent canonical or non-canonical autophagy pathways have been reported showing varying degrees of selectivity with regard to the substrates targeted. Here, we provide an updated review of the autophagy machinery and discuss the role of various forms of autophagy in neurodegenerative diseases, with a particular focus on Parkinson's disease. We describe recent findings that have led to the proposal of therapeutic strategies targeting autophagy to alter the course of Parkinson's disease progression.
    Keywords:  Parkinson’s disease; autoimmunity; autophagy; lysosomes; neurodegenerative disease
    DOI:  https://doi.org/10.3390/cells10123547
  4. Cells. 2021 Dec 17. pii: 3569. [Epub ahead of print]10(12):
    Mitophagy in Yeast: Molecular Mechanism and Regulation.
    Aleksei Innokentev, Tomotake Kanki.
      Mitophagy is a type of autophagy that selectively degrades mitochondria. Mitochondria, known as the "powerhouse of the cell", supply the majority of the energy required by cells. During energy production, mitochondria produce reactive oxygen species (ROS) as byproducts. The ROS damage mitochondria, and the damaged mitochondria further produce mitochondrial ROS. The increased mitochondrial ROS damage cellular components, including mitochondria themselves, and leads to diverse pathologies. Accordingly, it is crucial to eliminate excessive or damaged mitochondria to maintain mitochondrial homeostasis, in which mitophagy is believed to play a major role. Recently, the molecular mechanism and physiological role of mitophagy have been vigorously studied in yeast and mammalian cells. In yeast, Atg32 and Atg43, mitochondrial outer membrane proteins, were identified as mitophagy receptors in budding yeast and fission yeast, respectively. Here we summarize the molecular mechanisms of mitophagy in yeast, as revealed by the analysis of Atg32 and Atg43, and review recent progress in our understanding of mitophagy induction and regulation in yeast.
    Keywords:  Atg32; Atg43; autophagy; mitochondria; mitophagy; yeast
    DOI:  https://doi.org/10.3390/cells10123569
  5. Neurobiol Dis. 2021 Dec 18. pii: S0969-9961(21)00344-2. [Epub ahead of print] 105595
    Endophilin-B regulates autophagy during synapse development and neurodegeneration.
    Sergio Hernández-Díaz, Saurav Ghimire, Carla Montecinos-Oliva, Irene Sanchez-Mirasierra, Jef Swerts, Sabine Kuenen, Patrik Verstreken, Sandra-Fausia Soukup.
      Synapses are critical for neuronal communication and brain function. To maintain neuronal homeostasis, synapses rely on autophagy. Autophagic alterations cause neurodegeneration and synaptic dysfunction is a feature in neurodegenerative diseases. In Parkinson's disease (PD), where the loss of synapses precedes dopaminergic neuron loss, various PD-causative proteins are involved in the regulation of autophagy. So far only a few factors regulating autophagy at the synapse have been identified and the molecular mechanisms underlying autophagy at the synapse is only partially understood. Here, we describe Endophilin-B (EndoB) as a novel player in the regulation of synaptic autophagy in health and disease. We demonstrate that EndoB is required for autophagosome biogenesis at the synapse, whereas the loss of EndoB blocks the autophagy induction promoted by the PD mutation LRRK2G2019S. We show that EndoB is required to prevent neuronal loss. Moreover, loss of EndoB in the Drosophila visual system leads to an increase in synaptic contacts between photoreceptor terminals and their post-synaptic synapses. These data confirm the role of autophagy in synaptic contact formation and neuronal survival.
    Keywords:  Autophagy; Drosophila; Neurodegeneration; Parkinson's disease; Synapse development
    DOI:  https://doi.org/10.1016/j.nbd.2021.105595
  6. Autophagy. 2021 Dec 19. 1-17
    Autophagy modulates cell fate decisions during lineage commitment.
    Kulbhushan Sharma, Nagham T Asp, Sean P Harrison, Richard Siller, Saphira F Baumgarten, Swapnil Gupta, Maria E Chollet, Elisabeth Andersen, Gareth J Sullivan, Anne Simonsen.
      Early events during development leading to exit from a pluripotent state and commitment toward a specific germ layer still need in depth understanding. Autophagy has been shown to play a crucial role in both development and differentiation. This study employs human embryonic and induced pluripotent stem cells to understand the early events of lineage commitment with respect to the role of autophagy in this process. Our data indicate that a dip in autophagy facilitates exit from pluripotency. Upon exit, we demonstrate that the modulation of autophagy affects SOX2 levels and lineage commitment, with induction of autophagy promoting SOX2 degradation and mesendoderm formation, whereas inhibition of autophagy causes SOX2 accumulation and neuroectoderm formation. Thus, our results indicate that autophagy-mediated SOX2 turnover is a determining factor for lineage commitment. These findings will deepen our understanding of development and lead to improved methods to derive different lineages and cell types.Abbreviations: ACTB: Actin, beta; ATG: Autophagy-related; BafA1: Bafilomycin A1; CAS9: CRISPR associated protein 9; CQ: Chloroquine; DE: Definitive endoderm; hESCs: Human Embryonic Stem Cells; hiPSCs: Human Induced Pluripotent Stem Cells; LAMP1: Lysosomal Associated Membrane Protein 1; MAP1LC3: Microtubule-Associated Protein 1 Light Chain 3; MTOR: Mechanistic Target Of Rapamycin Kinase; NANOG: Nanog Homeobox; PAX6: Paired Box 6; PE: Phosphatidylethanolamine; POU5F1: POU class 5 Homeobox 1; PRKAA2: Protein Kinase AMP-Activated Catalytic Subunit Alpha 2; SOX2: SRY-box Transcription Factor 2; SQSTM1: Sequestosome 1; ULK1: unc-51 like Autophagy Activating Kinase 1; WDFY3: WD Repeat and FYVE Domain Containing 3.
    Keywords:  Autophagosome; SOX2; differentiation; ectoderm; endoderm; mesoderm; pluripotent stem cells
    DOI:  https://doi.org/10.1080/15548627.2021.2008691
  7. Front Cell Infect Microbiol. 2021 ;11 796664
    The Role of Chaperone-Mediated Autophagy in Hepatitis C Virus-Induced Pathogenesis.
    Chieko Matsui, Putu Yuliandari, Lin Deng, Takayuki Abe, Ikuo Shoji.
      Lysosome incorporate and degrade proteins in a process known as autophagy. There are three types of autophagy; macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). Although autophagy is considered a nonselective degradation process, CMA is known as a selective degradation pathway. All proteins internalized in the lysosome via CMA contain a pentapeptide KFERQ-motif, also known as a CMA-targeting motif, which is necessary for selectivity. CMA directly delivers a substrate protein into the lysosome lumen using the cytosolic chaperone HSC70 and the lysosomal receptor LAMP-2A for degradation. Hepatitis C virus (HCV) NS5A protein interacts with hepatocyte-nuclear factor 1α (HNF-1α) together with HSC70 and promotes the lysosomal degradation of HNF-1α via CMA, resulting in HCV-induced pathogenesis. HCV NS5A promotes recruitment of HSC70 to the substrate protein HNF-1α. HCV NS5A plays a crucial role in HCV-induced CMA. Further investigations of HCV NS5A-interacting proteins containing CMA-targeting motifs may help to elucidate HCV-induced pathogenesis.
    Keywords:  CMA-targeting motif; HSC70; LAMP-2A; chaperone-mediated autophagy; hepatitis C virus; lysosome
    DOI:  https://doi.org/10.3389/fcimb.2021.796664
  8. J Neuroinflammation. 2021 Dec 22. 18(1): 297
    Mitophagy in neurological disorders.
    Lijun Zhang, Lei Dai, Deyuan Li.
      Selective autophagy is an evolutionarily conserved mechanism that removes excess protein aggregates and damaged intracellular components. Most eukaryotic cells, including neurons, rely on proficient mitophagy responses to fine-tune the mitochondrial number and preserve energy metabolism. In some circumstances (such as the presence of pathogenic protein oligopolymers and protein mutations), dysfunctional mitophagy leads to nerve degeneration, with age-dependent intracellular accumulation of protein aggregates and dysfunctional organelles, leading to neurodegenerative disease. However, when pathogenic protein oligopolymers, protein mutations, stress, or injury are present, mitophagy prevents the accumulation of damaged mitochondria. Accordingly, mitophagy mediates neuroprotective effects in some forms of neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, and Amyotrophic lateral sclerosis) and acute brain damage (e.g., stroke, hypoxic-ischemic brain injury, epilepsy, and traumatic brain injury). The complex interplay between mitophagy and neurological disorders suggests that targeting mitophagy might be applicable for the treatment of neurodegenerative diseases and acute brain injury. However, due to the complexity of the mitophagy mechanism, mitophagy can be both harmful and beneficial, and future efforts should focus on maximizing its benefits. Here, we discuss the impact of mitophagy on neurological disorders, emphasizing the contrast between the positive and negative effects of mitophagy.
    Keywords:  Alzheimer's disease; Autophagy; Huntington's disease; Mitophagy; Neurological diseases; Stroke
    DOI:  https://doi.org/10.1186/s12974-021-02334-5
  9. Cells. 2021 Dec 15. pii: 3541. [Epub ahead of print]10(12):
    Mitophagy in Yeast: Decades of Research.
    Bhatia-Kissova Ingrid, Camougrand Nadine.
      Mitophagy, the selective degradation of mitochondria by autophagy, is one of the most important mechanisms of mitochondrial quality control, and its proper functioning is essential for cellular homeostasis. In this review, we describe the most important milestones achieved during almost 2 decades of research on yeasts, which shed light on the molecular mechanisms, regulation, and role of the Atg32 receptor in this process. We analyze the role of ROS in mitophagy and discuss the physiological roles of mitophagy in unicellular organisms, such as yeast; these roles are very different from those in mammals. Additionally, we discuss some of the different tools available for studying mitophagy.
    Keywords:  Atg32 protein; mitochondria; mitophagy; quality control; yeast
    DOI:  https://doi.org/10.3390/cells10123541
  10. J Neuroinflammation. 2021 Dec 20. 18(1): 295
    p38-TFEB pathways promote microglia activation through inhibiting CMA-mediated NLRP3 degradation in Parkinson's disease.
    Jialong Chen, Kanmin Mao, Honglin Yu, Yue Wen, Hua She, He Zhang, Linhua Liu, Mingque Li, Wenjun Li, Fei Zou.
       BACKGROUND: Parkinson's disease (PD) is characterized by degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNpc), accompanied by accumulation of α-synuclein, chronic neuroinflammation and autophagy dysfunction. Previous studies suggested that misfolded α-synuclein induces the inflammatory response and autophagy dysfunction in microglial cells. The NLRP3 inflammasome signaling pathway plays a crucial role in the neuroinflammatory process in the central nervous system. However, the relationship between autophagy deficiency and NLRP3 activation induced by α-synuclein accumulation is not well understood.
    METHODS: Through immunoblotting, immunocytochemistry, immunofluorescence, flow cytometry, ELISA and behavioral tests, we investigated the role of p38-TFEB-NLRP3 signaling pathways on neuroinflammation in the α-synuclein A53T PD models.
    RESULTS: Our results showed that increased protein levels of NLRP3, ASC, and caspase-1 in the α-synuclein A53T PD models. P38 is activated by overexpression of α-synuclein A53T mutant, which inhibited the master transcriptional activator of autophagy TFEB. And we found that NLRP3 was degraded by chaperone-mediated autophagy (CMA) in microglial cells. Furthermore, p38-TFEB pathways inhibited CMA-mediated NLRP3 degradation in Parkinson's disease. Inhibition of p38 had a protective effect on Parkinson's disease model via suppressing the activation of NLRP3 inflammasome pathway. Moreover, both p38 inhibitor SB203580 and NLRP3 inhibitor MCC950 not only prevented neurodegeneration in vivo, but also alleviated movement impairment in α-synuclein A53T-tg mice model of Parkinson's disease.
    CONCLUSION: Our research reveals p38-TFEB pathways promote microglia activation through inhibiting CMA-mediated NLRP3 degradation in Parkinson's disease, which could be a potential therapeutic strategy for PD. p38-TFEB pathways promote microglia activation through inhibiting CMA-mediated NLRP3 degradation in Parkinson's disease. In this model, p38 activates NLRP3 inflammasome via inhibiting TFEB in microglia. TFEB signaling negatively regulates NLRP3 inflammasome through increasing LAMP2A expression, which binds to NLRP3 and promotes its degradation via chaperone-mediated autophagy (CMA). NLRP3-mediated microglial activation promotes the death of dopaminergic neurons.
    Keywords:  Chaperone-mediated autophagy; NLRP3; Parkinson’s disease; TFEB; p38
    DOI:  https://doi.org/10.1186/s12974-021-02349-y
  11. Semin Cell Dev Biol. 2021 Dec 16. pii: S1084-9521(21)00308-6. [Epub ahead of print]
    Emerging role of mitophagy in myoblast differentiation and skeletal muscle remodeling.
    Fasih Ahmad Rahman, Joe Quadrilatero.
      Mitochondrial turnover in the form of mitophagy is emerging as a central process in maintaining cellular function. The degradation of damaged mitochondria through mitophagy is particularly important in cells/tissues that exhibit high energy demands. Skeletal muscle is one such tissue that requires precise turnover of mitochondria in several conditions in order to optimize energy production and prevent bioenergetic crisis. For instance, the formation of skeletal muscle (i.e., myogenesis) is accompanied by robust turnover of low-functioning mitochondria to eventually allow the formation of high-functioning mitochondria. In mature skeletal muscle, alterations in mitophagy-related signaling occur during exercise, aging, and various disease states. Nonetheless, several questions regarding the direct role of mitophagy in various skeletal muscle conditions remain unknown. Furthermore, given the heterogenous nature of skeletal muscle with respect to various cellular and molecular properties, and the plasticity in these properties in various conditions, the involvement and characterization of mitophagy requires more careful consideration in this tissue. Therefore, this review will highlight the known mechanisms of mitophagy in skeletal muscle, and discuss their involvement during myogenesis and various skeletal muscle conditions. This review also provides important considerations for the accurate measurement of mitophagy and interpretation of data in skeletal muscle.
    Keywords:  Aging; Atrophy; Autophagy; Cancer; Differentiation; Fiber type; Mitochondria; Mitochondrial network; Mitophagy; Myoblasts; Myogenesis; Regeneration; Remodeling; Skeletal muscle
    DOI:  https://doi.org/10.1016/j.semcdb.2021.11.026
  12. Cells. 2021 Dec 06. pii: 3432. [Epub ahead of print]10(12):
    Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy.
    Tonje Sønstevold, Nikolai Engedal, Maria Lyngaas Torgersen.
      Nanoparticles (NPs) are used in our everyday life, including as drug delivery vehicles. However, the effects of NPs at the cellular level and their impacts on autophagy are poorly understood. Here, we demonstrate that the NP drug delivery vehicle poly(butyl cyanoacrylate) (PBCA) perturbs redox homeostasis in human epithelial cells, and that the degree of redox perturbation dictates divergent effects of PBCA on autophagy. Specifically, PBCA promoted functional autophagy at low concentrations, whereas it inhibited autophagy at high concentrations. Both effects were completely abolished by the antioxidant N-acetyl cysteine (NAC). High concentrations of PBCA inhibited MAP1LC3B/GABARAP lipidation and LC3 flux, and blocked bulk autophagic cargo flux induced by mTOR inhibition. These effects were mimicked by the redox regulator H2O2. In contrast, low concentrations of PBCA enhanced bulk autophagic cargo flux in a Vps34-, ULK1/2- and ATG13-dependent manner, yet interestingly, without an accompanying increase in LC3 lipidation or flux. PBCA activated MAP kinase signaling cascades in a redox-dependent manner, and interference with individual signaling components revealed that the autophagy-stimulating effect of PBCA required the action of the JNK and p38-MK2 pathways, whose activities converged on the pro-autophagic protein Beclin-1. Collectively, our results reveal that PBCA exerts a dual effect on autophagy depending on the severity of the NP insult and the resulting perturbation of redox homeostasis. Such a dual autophagy-modifying effect may be of general relevance for redox-perturbing NPs and have important implications in nanomedicine.
    Keywords:  autophagy; nanoparticle; oxidative stress; p38; poly(alkyl cyanoacrylate)
    DOI:  https://doi.org/10.3390/cells10123432
  13. Mol Cells. 2021 Dec 15.
    Protein Kinase CK2 Is Upregulated by Calorie Restriction and Induces Autophagy.
    Jeong-Woo Park, Jihyeon Jeong, Young-Seuk Bae.
      Calorie restriction (CR) and the activation of autophagy extend healthspan by delaying the onset of age-associated diseases in most living organisms. Because protein kinase CK2 (CK2) downregulation induces cellular senescence and nematode aging, we investigated CK2's role in CR and autophagy. This study indicated that CR upregulated CK2's expression, thereby causing SIRT1 and AMP-activated protein kinase (AMPK) activation. CK2α overexpression, including antisense inhibitors of miR-186, miR-216b, miR-337-3p, and miR-760, stimulated autophagy initiation and nucleation markers (increase in ATG5, ATG7, LC3BII, beclin-1, and Ulk1, and decrease in SQSTM1/p62). The SIRT1 deacetylase, AKT, mammalian target of rapamycin (mTOR), AMPK, and forkhead homeobox type O (FoxO) 3a were involved in CK2-mediated autophagy. The treatment with the AKT inhibitor triciribine, the AMPK activator AICAR, or the SIRT1 activator resveratrol rescued a reduction in the expression of lgg-1 (the Caenorhabditis elegans ortholog of LC3B), bec-1 (the C. elegans ortholog of beclin-1), and unc-51 (the C. elegans ortholog of Ulk1), mediated by kin-10 (the C. elegans ortholog of CK2β) knockdown in nematodes. Thus, this study indicated that CK2 acted as a positive regulator in CR and autophagy, thereby suggesting that these four miRs' antisense inhibitors can be used as CR mimetics or autophagy inducers.
    Keywords:  CK2; SIRT1; autophagy; calorie restriction
    DOI:  https://doi.org/10.14348/molcells.2021.0183
  14. Cell Death Dis. 2021 Dec 20. 13(1): 14
    BNIP3L/NIX-mediated mitophagy: molecular mechanisms and implications for human disease.
    Yue Li, Wanqing Zheng, Yangyang Lu, Yanrong Zheng, Ling Pan, Xiaoli Wu, Yang Yuan, Zhe Shen, Shijia Ma, Xingxian Zhang, Jiaying Wu, Zhong Chen, Xiangnan Zhang.
      Mitophagy is a highly conserved cellular process that maintains the mitochondrial quantity by eliminating dysfunctional or superfluous mitochondria through autophagy machinery. The mitochondrial outer membrane protein BNIP3L/Nix serves as a mitophagy receptor by recognizing autophagosomes. BNIP3L is initially known to clear the mitochondria during the development of reticulocytes. Recent studies indicated it also engages in a variety of physiological and pathological processes. In this review, we provide an overview of how BNIP3L induces mitophagy and discuss the biological functions of BNIP3L and its regulation at the molecular level. We further discuss current evidence indicating the involvement of BNIP3L-mediated mitophagy in human disease, particularly in cancer and neurological disorders.
    DOI:  https://doi.org/10.1038/s41419-021-04469-y
  15. Front Cell Dev Biol. 2021 ;9 799123
    Mechanisms of FA-Phagy, a New Form of Selective Autophagy/Organellophagy.
    Jiayi Lu, Bernard Linares, Zhen Xu, Yan-Ning Rui.
      Focal adhesions (FAs) are adhesive organelles that attach cells to the extracellular matrix and can mediate various biological functions in response to different environmental cues. Reduced FAs are often associated with enhanced cell migration and cancer metastasis. In addition, because FAs are essential for preserving vascular integrity, the loss of FAs leads to hemorrhages and is frequently observed in many vascular diseases such as intracranial aneurysms. For these reasons, FAs are an attractive therapeutic target for treating cancer or vascular diseases, two leading causes of death world-wide. FAs are controlled by both their formation and turnover. In comparison to the large body of literature detailing FA formation, the mechanisms of FA turnover are poorly understood. Recently, autophagy has emerged as a major mechanism to degrade FAs and stabilizing FAs by inhibiting autophagy has a beneficial effect on breast cancer metastasis, suggesting autophagy-mediated FA turnover is a promising drug target. Intriguingly, autophagy-mediated FA turnover is a selective process and the cargo receptors for recognizing FAs in this process are context-dependent, which ensures the degradation of specific cargo. This paper mainly reviews the cargo recognition mechanisms of FA-phagy (selective autophagy-mediated FA turnover) and its disease relevance. We seek to outline some new points of understanding that will facilitate further study of FA-phagy and precise therapeutic strategies for related diseases associated with aberrant FA functions.
    Keywords:  autophagy; cancer; cargo receptor; focal adhesion; intracranial aneurysm; organellophagy; vascular integrity
    DOI:  https://doi.org/10.3389/fcell.2021.799123
  16. Cells. 2021 Dec 01. pii: 3389. [Epub ahead of print]10(12):
    Parkin beyond Parkinson's Disease-A Functional Meaning of Parkin Downregulation in TDP-43 Proteinopathies.
    Katarzyna Gaweda-Walerych, Emilia Jadwiga Sitek, Ewa Narożańska, Emanuele Buratti.
      Parkin and PINK1 are key regulators of mitophagy, an autophagic pathway for selective elimination of dysfunctional mitochondria. To this date, parkin depletion has been associated with recessive early onset Parkinson's disease (PD) caused by loss-of-function mutations in the PARK2 gene, while, in sporadic PD, the activity and abundance of this protein can be compromised by stress-related modifications. Intriguingly, research in recent years has shown that parkin depletion is not limited to PD but is also observed in other neurodegenerative diseases-especially those characterized by TDP-43 proteinopathies, such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Here, we discuss the evidence of parkin downregulation in these disease phenotypes, its emerging connections with TDP-43, and its possible functional implications.
    Keywords:  Parkinson’s disease (PD); TDP-43 pathology; amyotrophic lateral sclerosis (ALS); fronto-temporal lobar degeneration (FTLD); mitophagy; parkin
    DOI:  https://doi.org/10.3390/cells10123389
  17. Plant J. 2021 Dec 24.
    Phospholipase Dε Interacts with Autophagy-Related-Protein 8 and Promotes Autophagy in Arabidopsis Response to Nitrogen Deficiency.
    Shuaibing Yao, Shuming Peng, Xuemin Wang.
      Autophagy delivers cellular cargoes for degradation and plays an important role in nutrient recycling under nutrient limitation. A critical component in autophagy is the lipidation of phosphatidylethanolamine (PE) to the autophagy-related-protein 8, ATG8, to form ATG8-PE. Here we show that phospholipase Dε (PLDε) in Arabidopsis cells interacts with ATG8 and hydrolyzes ATG8-PE conjugates. In response to nitrogen deficiency, the transcript and protein levels of PLDε increase and additionally, an increasing amount of PLDε became associated with intracellular membranes compared with its primary plasma membrane association under sufficient nitrogen in Arabidopsis seedlings. PLDε-knockout (KO) plants have a lower, whereas PLDε-overexpressing (OE) ones have a higher number of autophagosomes than that in wild-type during nitrogen starvation. The level of ATG8-PE is lower in PLDε-KO, but higher in PLDε-OE plants than wild-type. PLDε-KO and -OE Arabidopsis seedlings display accelerated and delayed leaf senescence, respectively, during nitrogen limitation. The results suggest that PLDε promotes autophagy in plant response to nitrogen deficiency. The multifaceted effects of PLDε activity and interaction with ATG8 on autophagic processes are discussed.
    Keywords:  ATG8-PE conjugates; autophagy; lipid regulation; nitrogen responses; phospholipase D
    DOI:  https://doi.org/10.1111/tpj.15649
  18. Autophagy. 2021 Dec 19. 1-2
    Fuel for emotion: how mitophagy in the BLA can mediate increased anxiety in stressful social situations.
    James Johnson, Zheng Li.
      Psychosocial stress is a common risk factor for anxiety disorders. The cellular mechanism for the anxiogenic effect of psychosocial stress is largely unclear. We recently showed that chronic social defeat (CSD) stress in mice causes mitochondrial impairment, which triggers the PINK1-PRKN/parkin mitophagy pathway selectively in the amygdala. This mitophagy elevation causes excessive mitochondrial elimination and consequent mitochondrial deficiency. Mitochondrial deficiency in the basolateral amygdalae (BLA) causes weakening of synaptic transmission in the BLA-BNST (bed nucleus of the stria terminalis) anxiolytic pathway and increased anxiety. The CSD-induced increase in anxiety-like behaviors is abolished in pink1-/- and prkn-/- mice and alleviated by optogenetic activation of the BLA-BNST synapse. This study identifies an unsuspected role of mitophagy in psychogenetic-stress-induced anxiety elevation and reveals that mitochondrial deficiency is sufficient to increase anxiety and underlies the psychosocial-stress-induced anxiety increase. Mitochondria and mitophagy, therefore, can be potentially targeted to ameliorate anxiety.
    Keywords:  BNST; Mitophagy; amygdala; anxiety; mitochondria; psychological stress; social defeat stress
    DOI:  https://doi.org/10.1080/15548627.2021.2014769
  19. Autophagy. 2021 Dec 22. 1-3
    TFEB-dependent autophagy is involved in scavenger receptor OLR1/LOX-1-mediated tumor progression.
    Can Li, Lan Wang.
      Macroautophagy/autophagy is an evolutionarily conserved catabolic pathway required to maintain cellular homeostasis. In cancer, the tumor cell-intrinsic effects of autophagy are highly context specific, which could promote cancer cell survival or induce programmed cell death. Here, we reveal that OLR1/LOX-1 (oxidized low density lipoprotein receptor 1), a scavenger receptor highly expressed in esophageal cancer cells, is involved in tumorigenesis by suppressing autophagic cell death. Mechanistically, OLR1 binding to RACK1 activates MAP2K/MEK-MAPK/ERK signaling leading to TFEB (transcription factor EB) being trapped outside the nucleus and inhibiting autophagy. In addition, we identify a polysaccharide which causes the degradation of OLR1 and suppresses this autophagic pathway to inhibit tumorigenesis. This study demonstrates novel molecular mechanisms underlying the tumor-suppressive effect of autophagy and provides therapeutic insight for esophageal cancer.
    Keywords:  Cell death; ERK; RACK1; TFEB; fucoidan; tumorigenesis
    DOI:  https://doi.org/10.1080/15548627.2021.2012970
  20. Cells. 2021 Nov 27. pii: 3330. [Epub ahead of print]10(12):
    Molecular Signaling to Preserve Mitochondrial Integrity against Ischemic Stress in the Heart: Rescue or Remove Mitochondria in Danger.
    Justin D Yu, Shigeki Miyamoto.
      Cardiovascular diseases are one of the leading causes of death and global health problems worldwide, and ischemic heart disease is the most common cause of heart failure (HF). The heart is a high-energy demanding organ, and myocardial energy reserves are limited. Mitochondria are the powerhouses of the cell, but under stress conditions, they become damaged, release necrotic and apoptotic factors, and contribute to cell death. Loss of cardiomyocytes plays a significant role in ischemic heart disease. In response to stress, protective signaling pathways are activated to limit mitochondrial deterioration and protect the heart. To prevent mitochondrial death pathways, damaged mitochondria are removed by mitochondrial autophagy (mitophagy). Mitochondrial quality control mediated by mitophagy is functionally linked to mitochondrial dynamics. This review provides a current understanding of the signaling mechanisms by which the integrity of mitochondria is preserved in the heart against ischemic stress.
    Keywords:  BCL-2 family proteins; cell death; heart; ischemia; mPTP; mitochondria; mitophagy; pro-survival signaling
    DOI:  https://doi.org/10.3390/cells10123330
  21. Life Sci Alliance. 2022 Mar;pii: e202101185. [Epub ahead of print]5(3):
    Amyloid-like aggregating proteins cause lysosomal defects in neurons via gain-of-function toxicity.
    Irene Riera-Tur, Tillman Schäfer, Daniel Hornburg, Archana Mishra, Miguel da Silva Padilha, Lorena Fernández-Mosquera, Dennis Feigenbutz, Patrick Auer, Matthias Mann, Wolfgang Baumeister, Rüdiger Klein, Felix Meissner, Nuno Raimundo, Rubén Fernández-Busnadiego, Irina Dudanova.
      The autophagy-lysosomal pathway is impaired in many neurodegenerative diseases characterized by protein aggregation, but the link between aggregation and lysosomal dysfunction remains poorly understood. Here, we combine cryo-electron tomography, proteomics, and cell biology studies to investigate the effects of protein aggregates in primary neurons. We use artificial amyloid-like β-sheet proteins (β proteins) to focus on the gain-of-function aspect of aggregation. These proteins form fibrillar aggregates and cause neurotoxicity. We show that late stages of autophagy are impaired by the aggregates, resulting in lysosomal alterations reminiscent of lysosomal storage disorders. Mechanistically, β proteins interact with and sequester AP-3 μ1, a subunit of the AP-3 adaptor complex involved in protein trafficking to lysosomal organelles. This leads to destabilization of the AP-3 complex, missorting of AP-3 cargo, and lysosomal defects. Restoring AP-3μ1 expression ameliorates neurotoxicity caused by β proteins. Altogether, our results highlight the link between protein aggregation, lysosomal impairments, and neurotoxicity.
    DOI:  https://doi.org/10.26508/lsa.202101185
  22. J Biol Chem. 2021 Dec 17. pii: S0021-9258(21)01324-7. [Epub ahead of print] 101514
    An ALS-associated variant of the autophagy receptor SQSTM1/p62 reprograms binding selectivity towards the autophagy-related hATG8 proteins.
    Andrew Brennan, Robert Layfield, Jed Long, Huw E L Williams, Neil J Oldham, Daniel Scott, Mark S Searle.
      Recognition of human autophagy-related 8 (hATG8) proteins by autophagy receptors represents a critical step within this major cellular quality control system. Autophagy impairment is known to be a pathogenic mechanism in the motor neuron disorder amyotrophic lateral sclerosis (ALS). However, overlapping but specific roles of hATG8 proteins belonging to the LC3 and GABARAP subfamilies are incompletely understood, and binding selectivity is typically overlooked. We previously showed an ALS-associated variant of the SQSTM1/p62 (p62) autophagy receptor bearing an L341V mutation within its ATG8-interacting motif (AIM) impairs recognition of LC3B in vitro, yielding an autophagy-deficient phenotype. Improvements in understanding the molecular basis of hATG8 recognition by AIMs now distinguish LC3-interaction and GABARAP-interaction motifs and predict the effects of L341V substitution may extend beyond loss-of-function to biasing AIM binding preference. Through biophysical analyses, we confirm impaired binding of the L341V-AIM mutant to LC3A, LC3B, GABARAP, and GABARAPL1. In contrast, p62 AIM interactions with LC3C and GABARAPL2 are unaffected by this mutation. Isothermal titration calorimetry and NMR investigations provided insights into the entropy-driven GABARAPL2/p62 interaction and how the L341V mutation may be tolerated. Competition binding also demonstrated reduced association of the L341V-AIM with one hATG8 manifests as a relative increase in association with alternate hATG8s, indicating effective reprogramming of hATG8 selectivity. These data highlight how a single AIM peptide might compete for binding with different hATG8s, and suggest the L341V-AIM mutation may be neomorphic, representative of a disease mechanism that likely extends into other human disorders.
    Keywords:  AIM; SQSTM1/p62; amyotrophic lateral sclerosis; autophagy; hATG8
    DOI:  https://doi.org/10.1016/j.jbc.2021.101514
  23. PLoS Genet. 2021 Dec 23. 17(12): e1009980
    Formimidoyltransferase cyclodeaminase prevents the starvation-induced liver hepatomegaly and dysfunction through downregulating mTORC1.
    Wenfeng Zhang, Chaoying Wu, Rui Ni, Qifen Yang, Lingfei Luo, Jianbo He.
      The liver is a crucial center in the regulation of energy homeostasis under starvation. Although downregulation of mammalian target of rapamycin complex 1 (mTORC1) has been reported to play pivotal roles in the starvation responses, the underpinning mechanisms in particular upstream factors that downregulate mTORC1 remain largely unknown. To identify genetic variants that cause liver energy disorders during starvation, we conduct a zebrafish forward genetic screen. We identify a liver hulk (lvh) mutant with normal liver under feeding, but exhibiting liver hypertrophy under fasting. The hepatomegaly in lvh is caused by enlarged hepatocyte size and leads to liver dysfunction as well as limited tolerance to starvation. Positional cloning reveals that lvh phenotypes are caused by mutation in the ftcd gene, which encodes the formimidoyltransferase cyclodeaminase (FTCD). Further studies show that in response to starvation, the phosphorylated ribosomal S6 protein (p-RS6), a downstream effector of mTORC1, becomes downregulated in the wild-type liver, but remains at high level in lvh. Inhibition of mTORC1 by rapamycin rescues the hepatomegaly and liver dysfunction of lvh. Thus, we characterize the roles of FTCD in starvation response, which acts as an important upstream factor to downregulate mTORC1, thus preventing liver hypertrophy and dysfunction.
    DOI:  https://doi.org/10.1371/journal.pgen.1009980
  24. Cancers (Basel). 2021 Dec 07. pii: 6152. [Epub ahead of print]13(24):
    Emerging Role of Autophagy in the Development and Progression of Oral Squamous Cell Carcinoma.
    Yomna S Abd El-Aziz, Lionel Y W Leck, Patric J Jansson, Sumit Sahni.
      Autophagy is a cellular catabolic process, which is characterized by degradation of damaged proteins and organelles needed to supply the cell with essential nutrients. At basal levels, autophagy is important to maintain cellular homeostasis and development. It is also a stress responsive process that allows the cells to survive when subjected to stressful conditions such as nutrient deprivation. Autophagy has been implicated in many pathologies including cancer. It is well established that autophagy plays a dual role in different cancer types. There is emerging role of autophagy in oral squamous cell carcinoma (OSCC) development and progression. This review will focus on the role played by autophagy in relation to different aspects of cancer progression and discuss recent studies exploring the role of autophagy in OSCC. It will further discuss potential therapeutic approaches to target autophagy in OSCC.
    Keywords:  anti-cancer therapy; autophagy; autophagy inhibitors; cancer progression; oral squamous cell carcinoma
    DOI:  https://doi.org/10.3390/cancers13246152
  25. Front Pharmacol. 2021 ;12 747975
    HEXA-018, a Novel Inducer of Autophagy, Rescues TDP-43 Toxicity in Neuronal Cells.
    Shinrye Lee, Myungjin Jo, Hye Eun Lee, Yu-Mi Jeon, Seyeon Kim, Younghwi Kwon, Junghwa Woo, Shin Han, Ji Young Mun, Hyung-Jun Kim.
      The autophagy-lysosomal pathway is an essential cellular mechanism that degrades aggregated proteins and damaged cellular components to maintain cellular homeostasis. Here, we identified HEXA-018, a novel compound containing a catechol derivative structure, as a novel inducer of autophagy. HEXA-018 increased the LC3-I/II ratio, which indicates activation of autophagy. Consistent with this result, HEXA-018 effectively increased the numbers of autophagosomes and autolysosomes in neuronal cells. We also found that the activation of autophagy by HEXA-018 is mediated by the AMPK-ULK1 pathway in an mTOR-independent manner. We further showed that ubiquitin proteasome system impairment- or oxidative stress-induced neurotoxicity was significantly reduced by HEXA-018 treatment. Moreover, oxidative stress-induced mitochondrial dysfunction was strongly ameliorated by HEXA-018 treatment. In addition, we investigated the efficacy of HEXA-018 in models of TDP-43 proteinopathy. HEXA-018 treatment mitigated TDP-43 toxicity in cultured neuronal cell lines and Drosophila. Our data indicate that HEXA-018 could be a new drug candidate for TDP-43-associated neurodegenerative diseases.
    Keywords:  ALS; TDP-43; autophagy; catechol; mitochondrial dysfunction
    DOI:  https://doi.org/10.3389/fphar.2021.747975
  26. Front Cell Dev Biol. 2021 ;9 795685
    Protein Quality Control at the Mitochondrial Surface.
    Fabian den Brave, Arushi Gupta, Thomas Becker.
      Mitochondria contain two membranes, the outer and inner membrane. The outer membrane fulfills crucial functions for the communication of mitochondria with the cellular environment like exchange of lipids via organelle contact sites, the transport of metabolites and the formation of a signaling platform in apoptosis and innate immunity. The translocase of the outer membrane (TOM complex) forms the entry gate for the vast majority of precursor proteins that are produced on cytosolic ribosomes. Surveillance of the functionality of outer membrane proteins is critical for mitochondrial functions and biogenesis. Quality control mechanisms remove defective and mistargeted proteins from the outer membrane as well as precursor proteins that clog the TOM complex. Selective degradation of single proteins is also an important mode to regulate mitochondrial dynamics and initiation of mitophagy pathways. Whereas inner mitochondrial compartments are equipped with specific proteases, the ubiquitin-proteasome system is a central player in protein surveillance on the mitochondrial surface. In this review, we summarize our current knowledge about the molecular mechanisms that govern quality control of proteins at the outer mitochondrial membrane.
    Keywords:  Cdc48; TOM complex; mitochondria; protein quality control; protein sorting
    DOI:  https://doi.org/10.3389/fcell.2021.795685
  27. Mol Microbiol. 2021 Dec 20.
    Impairment of lysosomal function by Clostridioides difficile TcdB.
    Carmen Klepka, Moritz Sandmann, Helma Tatge, Matthew Mangan, Annabel Arens, Daniel Henkel, Ralf Gerhard.
      TcdB is a potent cytotoxin produced by pathogenic Clostridioides difficile that inhibits Rho GTPases by mono-glucosylation. TcdB enters cells via receptor-mediated endocytosis. The pathogenic glucosyltransferase domain (GTD) egresses endosomes by pH-mediated conformational changes, and is subsequently released in an autoproteolytic manner. We here investigated the uptake, localization and degradation of TcdB. TcdB colocalized with lysosomal marker protein LAMP1, verifying the endosomal-lysosomal route of the toxin. In pulse assays endocytosed TcdB declined to a limit of detection within two hours, whereas the released GTD accumulated for up to eight hours. We observed that autoproteolytic deficient TcdB NXN C698S was degraded significantly faster than wildtype TcdB, suggesting interference of TcdB with lysosomal degradation process. In fact, TcdB reduced lysosomal degradation of endosome cargo as tested with DQ-Green BSA. Lysosomal dysfunction was accompanied by perinuclear accumulation of LAMP1 and a weaker detection in immunoblots. Galectin-8 or galectin-3 were not recruited to lysosomes speaking against lysosome membrane damage. Changes in the autophagosomal marker LC3B, suggested additional indirect effect of lysosomal dysfunction on the autophagic flux. In contrast to necrotic signaling induced in by TcdB lysosomal dysfunction was not abolished by calcium channel blocker nifedipin, indicating separate cytopathogenic effects induced by TcdB during endo-lysosomal trafficking.
    Keywords:  Clostridioides difficile Toxin B; DQ-BSA assay; autophagic flux; lysosome inhibition
    DOI:  https://doi.org/10.1111/mmi.14864
  28. Nature. 2021 Dec 21.
    Activation mechanism of PINK1.
    Zhong Yan Gan, Sylvie Callegari, Simon A Cobbold, Thomas R Cotton, Michael J Mlodzianoski, Alexander F Schubert, Niall D Geoghegan, Kelly L Rogers, Andrew Leis, Grant Dewson, Alisa Glukhova, David Komander.
      Mutations in the protein kinase PINK1 lead to defects in mitophagy and cause autosomal recessive early onset Parkinson's Disease (EOPD)1,2. PINK1 has many unique features that enable it to phosphorylate ubiquitin and the ubiquitin-like domain of Parkin3-9. Structural analysis of PINK1 from diverse insect species10-12 with and without ubiquitin provided snapshots of distinct structural states yet did not explain how PINK1 is activated. We here elucidate the activation mechanism of PINK1 by crystallography and cryo-EM. A crystal structure of unphosphorylated Pediculus humanus corporis (Ph) PINK1 resolves a previously omitted N-terminal helix revealing how unphosphorylated yet active PINK1 is oriented on mitochondria. We further reveal a 2.35 Å cryo-EM structure of a symmetric PhPINK1 dimer trapped during the process of trans-autophosphorylation, and a 3.1 Å cryo-EM structure of phosphorylated PhPINK1 in the process of undergoing a conformational change to become an active ubiquitin kinase. Structures and phosphorylation studies further identify a role for regulatory PINK1 oxidation. Together, our work delineates the complete activation mechanism of PINK1, illuminates how PINK1 interacts with the mitochondrial outer membrane, and reveals how PINK1 activity may be modulated by mitochondrial reactive oxygen species.
    DOI:  https://doi.org/10.1038/s41586-021-04340-2
  29. Brain. 2021 Dec 22. pii: awab473. [Epub ahead of print]
    Distinct subcellular autophagy impairments in induced neurons from Huntington's disease patients.
    Karolina Pircs, Janelle Drouin-Ouellet, Vivien Horváth, Jeovanis Gil, Melinda Rezeli, Raquel Garza, Daniela A Grassi, Yogita Sharma, Isabelle St-Amour, Kate Harris, Marie E Jönsson, Pia A Johansson, Romina Vuono, Shaline V Fazal, Thomas Stoker, Bob A Hersbach, Kritika Sharma, Jessica Lagerwall, Stina Lagerström, Petter Storm, Sébastien S Hébert, György Marko-Varga, Malin Parmar, Roger A Barker, Johan Jakobsson.
      Huntington's disease (HD) is a neurodegenerative disorder caused by CAG expansions in the huntingtin (HTT) gene. Modelling Huntington's disease is challenging, as rodent and cellular models poorly recapitulate the disease as seen in aging humans. To address this, we generated induced neurons (iNs) through direct reprogramming of human skin fibroblasts, which retain age-dependent epigenetic characteristics. HD-iNs displayed profound deficits in autophagy, characterised by reduced transport of late autophagic structures from the neurites to the soma. These neurite-specific alterations in autophagy resulted in shorter, thinner and fewer neurites specifically in HD-iNs. CRISPRi-mediated silencing of HTT did not rescue this phenotype but rather resulted in additional autophagy alterations in ctrl-iNs, highlighting the importance of wild type HTT in normal neuronal autophagy. In summary, our work identifies a distinct subcellular autophagy impairment in adult patient derived Huntington's disease neurons and provides a new rational for future development of autophagy activation therapies.
    Keywords:  CRISPR interference; Huntington’s disease; autophagy; direct neural reprogramming; lentiviral vector
    DOI:  https://doi.org/10.1093/brain/awab473
  30. Cells. 2021 Nov 28. pii: 3337. [Epub ahead of print]10(12):
    Protein Aggregation in the ER: Calm behind the Storm.
    Haisen Li, Shengyi Sun.
      As one of the largest organelles in eukaryotic cells, the endoplasmic reticulum (ER) plays a vital role in the synthesis, folding, and assembly of secretory and membrane proteins. To maintain its homeostasis, the ER is equipped with an elaborate network of protein folding chaperones and multiple quality control pathways whose cooperative actions safeguard the fidelity of protein biogenesis. However, due to genetic abnormalities, the error-prone nature of protein folding and assembly, and/or defects or limited capacities of the protein quality control systems, nascent proteins may become misfolded and fail to exit the ER. If not cleared efficiently, the progressive accumulation of misfolded proteins within the ER may result in the formation of toxic protein aggregates, leading to the so-called "ER storage diseases". In this review, we first summarize our current understanding of the protein folding and quality control networks in the ER, including chaperones, unfolded protein response (UPR), ER-associated protein degradation (ERAD), and ER-selective autophagy (ER-phagy). We then survey recent research progress on a few ER storage diseases, with a focus on the role of ER quality control in the disease etiology, followed by a discussion on outstanding questions and emerging concepts in the field.
    Keywords:  ER; ER storage disease; ER-associated protein degradation; ER-phagy; chaperone; protein aggregate; unfolded protein response
    DOI:  https://doi.org/10.3390/cells10123337
  31. Biomolecules. 2021 Dec 02. pii: 1814. [Epub ahead of print]11(12):
    RalA, PLD and mTORC1 Are Required for Kinase-Independent Pathways in DGKβ-Induced Neurite Outgrowth.
    Takuya Kano, Ryosuke Tsumagari, Akio Nakashima, Ushio Kikkawa, Shuji Ueda, Minoru Yamanoue, Nobuyuki Takei, Yasuhito Shirai.
      Diacylglycerol kinase β (DGKβ) is an enzyme that converts diacylglycerol to phosphatidic acid and is mainly expressed in the cerebral cortex, hippocampus and striatum. We previously reported that DGKβ induces neurite outgrowth and spinogenesis, contributing to higher brain functions, including emotion and memory. To elucidate the mechanisms involved in neuronal development by DGKβ, we investigated the importance of DGKβ activity in the induction of neurite outgrowth using human neuroblastoma SH-SY5Y cells. Interestingly, both wild-type DGKβ and the kinase-negative (KN) mutant partially induced neurite outgrowth, and these functions shared a common pathway via the activation of mammalian target of rapamycin complex 1 (mTORC1). In addition, we found that DGKβ interacted with the small GTPase RalA and that siRNA against RalA and phospholipase D (PLD) inhibitor treatments abolished DGKβKN-induced neurite outgrowth. These results indicate that binding of RalA and activation of PLD and mTORC1 are involved in DGKβKN-induced neurite outgrowth. Taken together with our previous reports, mTORC1 is a key molecule in both kinase-dependent and kinase-independent pathways of DGKβ-mediated neurite outgrowth, which is important for higher brain functions.
    Keywords:  RalA; diacylglycerol kinase; mammalian target of rapamycin; neurite; phosphatidic acid; phospholipase D (PLD)
    DOI:  https://doi.org/10.3390/biom11121814
  32. J Cell Biol. 2022 Feb 07. pii: e202105060. [Epub ahead of print]221(2):
    CRISPR screens for lipid regulators reveal a role for ER-bound SNX13 in lysosomal cholesterol export.
    Albert Lu, Frank Hsieh, Bikal R Sharma, Sydney R Vaughn, Carlos Enrich, Suzanne R Pfeffer.
      We report here two genome-wide CRISPR screens performed to identify genes that, when knocked out, alter levels of lysosomal cholesterol or bis(monoacylglycero)phosphate. In addition, these screens were also performed under conditions of NPC1 inhibition to identify modifiers of NPC1 function in lysosomal cholesterol export. The screens confirm tight coregulation of cholesterol and bis(monoacylglycero)phosphate in cells and reveal an unexpected role for the ER-localized SNX13 protein as a negative regulator of lysosomal cholesterol export and contributor to ER-lysosome membrane contact sites. In the absence of NPC1 function, SNX13 knockdown redistributes lysosomal cholesterol and is accompanied by triacylglycerol-rich lipid droplet accumulation and increased lysosomal bis(monoacylglycero)phosphate. These experiments provide unexpected insight into the regulation of lysosomal lipids and modification of these processes by novel gene products.
    DOI:  https://doi.org/10.1083/jcb.202105060
  33. Cells. 2021 Dec 15. pii: 3545. [Epub ahead of print]10(12):
    Activation of Downstream mTORC1 Target Ribosomal Protein S6 Kinase (S6K) Can Be Found in a Subgroup of Dutch Patients with Granulomatous Pulmonary Disease.
    Raisa Kraaijvanger, Kees Seldenrijk, Els Beijer, Jan Damen, Jayne Louise Wilson, Thomas Weichhart, Jan C Grutters, Marcel Veltkamp.
      Mechanistic target of rapamycin complex 1 (mTORC1) has been linked to different diseases. The mTORC1 signaling pathway is suggested to play a role in the granuloma formation of sarcoidosis. Recent studies demonstrated conflicting data on mTORC1 activation in patients with sarcoidosis by measuring activation of its downstream target S6 kinase (S6K) with either 33% or 100% of patients. Therefore, the aim of our study was to reevaluate the percentage of S6K activation in sarcoidosis patients in a Dutch cohort. To investigate whether this activation is specific for sarcoid granulomas, we also included Dutch patients with other granulomatous diseases of the lung. The activation of the S6K signaling pathway was evaluated by immunohistochemical staining of its downstream effector phospho-S6 in tissue sections. Active S6K signaling was detected in 32 (43%) of the sarcoidosis patients. Twelve (31%) of the patients with another granulomatous disorder also showed activated S6K signaling, demonstrating that the mTORC1 pathway may be activated in a range for different granulomatous diseases (p = 0.628). Activation of S6K can only be found in a subgroup of patients with sarcoidosis, as well as in patients with other granulomatous pulmonary diseases, such as hypersensitivity pneumonitis or vasculitis. No association between different clinical phenotypes and S6K activation can be found in sarcoidosis.
    Keywords:  S6K; granuloma; mTORC1; phenotyping; sarcoidosis
    DOI:  https://doi.org/10.3390/cells10123545
  34. Exp Cell Res. 2021 Dec 20. pii: S0014-4827(21)00542-5. [Epub ahead of print] 112986
    The two-domain architecture of LAMP2A regulates its interaction with Hsc70.
    Yuta Ikami, Kazue Terasawa, Kensaku Sakamoto, Kazumasa Ohtake, Hiroyuki Harada, Tetsuro Watabe, Shigeyuki Yokoyama, Miki Hara-Yokoyama.
      Chaperone-mediated autophagy (CMA) is a unique proteolytic pathway, in which cytoplasmic proteins recognized by heat shock cognate protein 70 (Hsc70/HSPA8) are transported into lysosomes for degradation. The substrate/chaperone complex binds to the cytosolic tail of the lysosomal-associated membrane protein type 2A (LAMP2A), but whether the interaction between Hsc70 and LAMP2A is direct or mediated by other molecules has remained to be elucidated. The structure of LAMP2A comprises a large lumenal domain composed of two domains, both with the β-prism fold, a transmembrane domain and a short cytoplasmic tail. We previously reported the structural basis for the homophilic interaction of the lumenal domains of LAMP2A, using site-specific photo-crosslinking and/or steric hindrance within cells. In the present study, we introduced a photo-crosslinker into the cytoplasmic tail of LAMP2A and successfully detected its crosslinking with Hsc70, revealing this direct interaction for the first time. Furthermore, we demonstrated that the truncation of the membrane-distal domain within the lumenal domain of LAMP2A reduced the amount of Hsc70 that coimmunoprecipitated with LAMP2A. Our present results suggested that the two-domain architecture of the lumenal domains of LAMP2A underlies the interaction with Hsc70 at the cytoplasmic surface of the lysosome.
    Keywords:  Chaperone-mediated autophagy; Expanded genetic codes; Lysosomes; Photo-crosslinking; Protein assembly
    DOI:  https://doi.org/10.1016/j.yexcr.2021.112986
  35. Int J Mol Sci. 2021 Dec 14. pii: 13447. [Epub ahead of print]22(24):
    Identification of Novel Therapeutic Targets for Polyglutamine Diseases That Target Mitochondrial Fragmentation.
    Annika Traa, Emily Machiela, Paige D Rudich, Sonja K Soo, Megan M Senchuk, Jeremy M Van Raamsdonk.
      Huntington's disease (HD) is one of at least nine polyglutamine diseases caused by a trinucleotide CAG repeat expansion, all of which lead to age-onset neurodegeneration. Mitochondrial dynamics and function are disrupted in HD and other polyglutamine diseases. While multiple studies have found beneficial effects from decreasing mitochondrial fragmentation in HD models by disrupting the mitochondrial fission protein DRP1, disrupting DRP1 can also have detrimental consequences in wild-type animals and HD models. In this work, we examine the effect of decreasing mitochondrial fragmentation in a neuronal C. elegans model of polyglutamine toxicity called Neur-67Q. We find that Neur-67Q worms exhibit mitochondrial fragmentation in GABAergic neurons and decreased mitochondrial function. Disruption of drp-1 eliminates differences in mitochondrial morphology and rescues deficits in both movement and longevity in Neur-67Q worms. In testing twenty-four RNA interference (RNAi) clones that decrease mitochondrial fragmentation, we identified eleven clones-each targeting a different gene-that increase movement and extend lifespan in Neur-67Q worms. Overall, we show that decreasing mitochondrial fragmentation may be an effective approach to treating polyglutamine diseases and we identify multiple novel genetic targets that circumvent the potential negative side effects of disrupting the primary mitochondrial fission gene drp-1.
    Keywords:  C. elegans; DRP1; Huntington’s disease; genetics; mitochondria; mitochondrial dynamics; polyglutamine diseases
    DOI:  https://doi.org/10.3390/ijms222413447
  36. ACS Pharmacol Transl Sci. 2021 Dec 10. 4(6): 1728-1746
    Cell Survival and Cell Death at the Intersection of Autophagy and Apoptosis: Implications for Current and Future Cancer Therapeutics.
    Nicole Bata, Nicholas D P Cosford.
      Autophagy and apoptosis are functionally distinct mechanisms for cytoplasmic and cellular turnover. While these two pathways are distinct, they can also regulate each other, and central components of the apoptosis or autophagy pathway regulate both processes directly. Furthermore, several upstream stress-inducing signaling pathways can influence both autophagy and apoptosis. The crosstalk between autophagy and apoptosis has an integral role in pathological processes, including those related to cancer, homeostasis, and aging. Apoptosis is a form of programmed cell death, tightly regulated by various cellular and biochemical mechanisms, some of which have been the focus of drug discovery efforts targeting cancer therapeutics. Autophagy is a cellular degradation pathway whereby cells recycle macromolecules and organelles to generate energy when subjected to stress. Autophagy can act as either a prodeath or a prosurvival process and is both tissue and microenvironment specific. In this review we describe five groups of proteins that are integral to the apoptosis pathway and discuss their role in regulating autophagy. We highlight several apoptosis-inducing small molecules and biologics that have been developed and advanced into the clinic and discuss their effects on autophagy. For the most part, these apoptosis-inducing compounds appear to elevate autophagy activity. Under certain circumstances autophagy demonstrates cytoprotective functions and is overactivated in response to chemo- or radiotherapy which can lead to drug resistance, representing a clinical obstacle for successful cancer treatment. Thus, targeting the autophagy pathway in combination with apoptosis-inducing compounds may be a promising strategy for cancer therapy.
    DOI:  https://doi.org/10.1021/acsptsci.1c00130
  37. Int J Mol Sci. 2021 Dec 15. pii: 13455. [Epub ahead of print]22(24):
    Apigenin Induces Autophagy and Cell Death by Targeting EZH2 under Hypoxia Conditions in Gastric Cancer Cells.
    Tae Woo Kim, Hee Gu Lee.
      Hypoxia is a major obstacle to gastric cancer (GC) therapy and leads to chemoresistance as GC cells are frequently exposed to the hypoxia environment. Apigenin, a flavonoid found in traditional medicine, fruits, and vegetables and an HDAC inhibitor, is a powerful anti-cancer agent against various cancer cell lines. However, detailed mechanisms involved in the treatment of GC using APG are not fully understood. In this study, we investigated the biological activity of and molecular mechanisms involved in APG-mediated treatment of GC under hypoxia. APG promoted autophagic cell death by increasing ATG5, LC3-II, and phosphorylation of AMPK and ULK1 and down-regulating p-mTOR and p62 in GC. Furthermore, our results show that APG induces autophagic cell death via the activation of the PERK signaling, indicating an endoplasmic reticulum (ER) stress response. The inhibition of ER stress suppressed APG-induced autophagy and conferred prolonged cell survival, indicating autophagic cell death. We further show that APG induces ER stress- and autophagy-related cell death through the inhibition of HIF-1α and Ezh2 under normoxia and hypoxia. Taken together, our findings indicate that APG activates autophagic cell death by inhibiting HIF-1α and Ezh2 under hypoxia conditions in GC cells.
    Keywords:  ER stress; apigenin; autophagy; hypoxia; resistance
    DOI:  https://doi.org/10.3390/ijms222413455
  38. Biochem Biophys Res Commun. 2021 Dec 10. pii: S0006-291X(21)01654-5. [Epub ahead of print]589 247-253
    Targeting FAM134B-mediated reticulophagy activates sorafenib-induced ferroptosis in hepatocellular carcinoma.
    Zhiqian Liu, Changlin Ma, Qi Wang, Hao Yang, Zhihua Lu, Tao Bi, Zongzhen Xu, Tao Li, Ling Zhang, Yajie Zhang, Jingfang Liu, Xiaoqing Wei, Jie Li.
      Ferroptosis is a kind of cell death closely related to selective autophagy, such as ferritinophagy, lipophagy, clockophagy and chaperone-mediated autophagy. However, the role of reticulophagy, which specifically degrades endoplasmic reticulum (ER) fragments (also known as ER-phagy), in ferroptosis regulation is still unclear. In this study, we found that sorafenib (ferroptosis inducer) can effectively activate the receptor protein FAM134B-mediated ER-phagy, and FAM134B knockdown not only blocked ER-phagy but also significantly strengthened cellular sensitivity to ferroptosis without affecting macroautophagy. In vivo experiments also yielded similar results. These evidences provided new clues for ferroptosis regulation. Subsequently, bioinformatic analysis combined with RNA binding protein immunoprecipitation and polyribosome fractionation preliminarily indicated that PABPC1 can interact with FAM134B mRNA and promote its translation. Taken together, this study revealed the role of the PABPC1-FAM134B-ER-phagy pathway on ferroptosis, providing important evidence for novel anti-cancer strategies.
    Keywords:  FAM134B; Ferroptosis; PABPC1; Reticulophagy/ER-Phagy
    DOI:  https://doi.org/10.1016/j.bbrc.2021.12.019
  39. Biomedicines. 2021 Dec 01. pii: 1809. [Epub ahead of print]9(12):
    The Bumpy Road towards mTOR Inhibition in Glioblastoma: Quo Vadis?
    Kostas A Papavassiliou, Athanasios G Papavassiliou.
      Glioblastoma multiforme (GBM), a grade IV astrocytoma, is a lethal brain tumor with a poor prognosis. Despite recent advances in the molecular biology of GBM, neuro-oncologists have very limited treatment options available to improve the survival of GBM patients. A prominent signaling pathway implicated in GBM pathogenesis is that of the mechanistic target of rapamycin (mTOR). Attempts to target the mTOR pathway with first-generation mTOR inhibitors appeared promising in the preclinical stage; however, results have been disappointing in clinical trials, owing to the heterogeneous nature of GBM, escape mechanisms against treatment, the blood-brain barrier, drug-related toxicities, and the imperfect design of clinical trials, among others. The development of next-generation mTOR inhibitors and their current evaluation in clinical trials have sparked new hope to realize the clinical potential of mTOR inhibitors in GBM. Meanwhile, studies are continuously furthering our understanding of mTOR signaling dysregulation, its downstream effects, and interplay with other signaling pathways in GBM tumors. Therefore, it remains to be seen whether targeting mTOR in GBM will eventually prove to be fruitful or futile.
    Keywords:  glioblastoma multiforme; mTOR inhibitors; mTOR signaling; mTORC1; mTORC2
    DOI:  https://doi.org/10.3390/biomedicines9121809
  40. Int J Mol Sci. 2021 Dec 08. pii: 13232. [Epub ahead of print]22(24):
    How the Innate Immune DNA Sensing cGAS-STING Pathway Is Involved in Autophagy.
    Wanglong Zheng, Nengwen Xia, Jiajia Zhang, Nanhua Chen, François Meurens, Zongping Liu, Jianzhong Zhu.
      The cGAS-STING pathway is a key component of the innate immune system and exerts crucial roles in the detection of cytosolic DNA and invading pathogens. Accumulating evidence suggests that the intrinsic cGAS-STING pathway not only facilitates the production of type I interferons (IFN-I) and inflammatory responses but also triggers autophagy. Autophagy is a homeostatic process that exerts multiple effects on innate immunity. However, systematic evidence linking the cGAS-STING pathway and autophagy is still lacking. Therefore, one goal of this review is to summarize the known mechanisms of autophagy induced by the cGAS-STING pathway and their consequences. The cGAS-STING pathway can trigger canonical autophagy through liquid-phase separation of the cGAS-DNA complex, interaction of cGAS and Beclin-1, and STING-triggered ER stress-mTOR signaling. Furthermore, both cGAS and STING can induce non-canonical autophagy via LC3-interacting regions and binding with LC3. Subsequently, autophagy induced by the cGAS-STING pathway plays crucial roles in balancing innate immune responses, maintaining intracellular environmental homeostasis, alleviating liver injury, and limiting tumor growth and transformation.
    Keywords:  DNA sensing; IFN; STING; autophagy; cGAS; innate immunity
    DOI:  https://doi.org/10.3390/ijms222413232
  41. Cancers (Basel). 2021 Dec 16. pii: 6328. [Epub ahead of print]13(24):
    The Autophagic Route of E-Cadherin and Cell Adhesion Molecules in Cancer Progression.
    Manuela Santarosa, Roberta Maestro.
      Cell-to-cell adhesion is a key element in epithelial tissue integrity and homeostasis during embryogenesis, response to damage, and differentiation. Loss of cell adhesion and gain of mesenchymal features, a phenomenon known as epithelial to mesenchymal transition (EMT), are essential steps in cancer progression. Interestingly, downregulation or degradation by endocytosis of epithelial adhesion molecules (e.g., E-cadherin) associates with EMT and promotes cell migration. Autophagy is a physiological intracellular degradation and recycling process. In cancer, it is thought to exert a tumor suppressive role in the early phases of cell transformation but, once cells have gained a fully transformed phenotype, autophagy may fuel malignant progression by promoting EMT and conferring drug resistance. In this review, we discuss the crosstalk between autophagy, EMT, and turnover of epithelial cell adhesion molecules, with particular attention to E-cadherin.
    Keywords:  E-cadherin; adherens junctions; autophagy; cancer; carcinoma; cell adhesion; epithelial to mesenchymal transition; metastasis
    DOI:  https://doi.org/10.3390/cancers13246328
  42. Cells. 2021 Nov 26. pii: 3319. [Epub ahead of print]10(12):
    Aging of Podospora anserina Leads to Alterations of OXPHOS and the Induction of Non-Mitochondrial Salvage Pathways.
    Verena Warnsmann, Jana Meisterknecht, Ilka Wittig, Heinz D Osiewacz.
      The accumulation of functionally impaired mitochondria is a key event in aging. Previous works with the fungal aging model Podospora anserina demonstrated pronounced age-dependent changes of mitochondrial morphology and ultrastructure, as well as alterations of transcript and protein levels, including individual proteins of the oxidative phosphorylation (OXPHOS). The identified protein changes do not reflect the level of the whole protein complexes as they function in-vivo. In the present study, we investigated in detail the age-dependent changes of assembled mitochondrial protein complexes, using complexome profiling. We observed pronounced age-depen-dent alterations of the OXPHOS complexes, including the loss of mitochondrial respiratory supercomplexes (mtRSCs) and a reduction in the abundance of complex I and complex IV. Additionally, we identified a switch from the standard complex IV-dependent respiration to an alternative respiration during the aging of the P. anserina wild type. Interestingly, we identified proteasome components, as well as endoplasmic reticulum (ER) proteins, for which the recruitment to mitochondria appeared to be increased in the mitochondria of older cultures. Overall, our data demonstrate pronounced age-dependent alterations of the protein complexes involved in energy transduction and suggest the induction of different non-mitochondrial salvage pathways, to counteract the age-dependent mitochondrial impairments which occur during aging.
    Keywords:  OXPHOS; Podospora anserina; aging; complexome profiling
    DOI:  https://doi.org/10.3390/cells10123319
  43. Int J Mol Sci. 2021 Dec 13. pii: 13398. [Epub ahead of print]22(24):
    Ellagic Acid Prevents α-Synuclein Aggregation and Protects SH-SY5Y Cells from Aggregated α-Synuclein-Induced Toxicity via Suppression of Apoptosis and Activation of Autophagy.
    Mustafa T Ardah, Nabil Eid, Tohru Kitada, M Emdadul Haque.
      Parkinson's disease (PD) is a neurodegenerative disease characterized by the loss of dopamine neurons and the deposition of misfolded proteins known as Lewy bodies (LBs), which contain α-synuclein (α-syn). The causes and molecular mechanisms of PD are not clearly understood to date. However, misfolded proteins, oxidative stress, and impaired autophagy are believed to play important roles in the pathogenesis of PD. Importantly, α-syn is considered a key player in the development of PD. The present study aimed to assess the role of Ellagic acid (EA), a polyphenol found in many fruits, on α-syn aggregation and toxicity. Using thioflavin and seeding polymerization assays, in addition to electron microscopy, we found that EA could dramatically reduce α-syn aggregation. Moreover, EA significantly mitigated the aggregated α-syn-induced toxicity in SH-SY5Y cells and thus enhanced their viability. Mechanistically, these cytoprotective effects of EA are mediated by the suppression of apoptotic proteins BAX and p53 and a concomitant increase in the anti-apoptotic protein, BCL-2. Interestingly, EA was able to activate autophagy in SH-SY5Y cells, as evidenced by normalized/enhanced expression of LC3-II, p62, and pAKT. Together, our findings suggest that EA may attenuate α-syn toxicity by preventing aggregation and improving viability by restoring autophagy and suppressing apoptosis.
    Keywords:  Ellagic acid; Parkinson’s disease; neurodegeneration; α-synuclein
    DOI:  https://doi.org/10.3390/ijms222413398
  44. Antioxidants (Basel). 2021 Nov 29. pii: 1917. [Epub ahead of print]10(12):
    ROS-Induced mtDNA Release: The Emerging Messenger for Communication between Neurons and Innate Immune Cells during Neurodegenerative Disorder Progression.
    Yuanxin Zhao, Buhan Liu, Long Xu, Sihang Yu, Jiaying Fu, Jian Wang, Xiaoyu Yan, Jing Su.
      One of the most striking hallmarks shared by various neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis, is microglia-mediated and astrocyte-mediated neuroinflammation. Although inhibitions of both harmful proteins and aggregation are major treatments for neurodegenerative diseases, whether the phenomenon of non-normal protein or peptide aggregation is causally related to neuronal loss and synaptic damage is still controversial. Currently, excessive production of reactive oxygen species (ROS), which induces mitochondrial dysfunction in neurons that may play a key role in the regulation of immune cells, is proposed as a regulator in neurological disorders. In this review, we propose that mitochondrial DNA (mtDNA) release due to ROS may act on microglia and astrocytes adjacent to neurons to induce inflammation through activation of innate immune responses (such as cGAS/STING). Elucidating the relationship between mtDNA and the formation of a pro-inflammatory microenvironment could contribute to a better understanding of the mechanism of crosstalk between neuronal and peripheral immune cells and lead to the development of novel therapeutic approaches to neurodegenerative diseases.
    Keywords:  ROS; cGAS/STING; mtDNA; neurodegenerative diseases; neuroinflammation
    DOI:  https://doi.org/10.3390/antiox10121917
  45. Cell Death Dis. 2021 Dec 20. 13(1): 13
    MAP4K4 mediates the SOX6-induced autophagy and reduces the chemosensitivity of cervical cancer.
    Hongxin Huang, Qin Han, Han Zheng, Mingchen Liu, Shu Shi, Ting Zhang, Xingwen Yang, Zhongqing Li, Qiang Xu, Hongyan Guo, Fengmin Lu, Jie Wang.
      There are nearly 40% of cervical cancer patients showing poor response to neoadjuvant chemotherapy that can be induced by autophagy, however, the underlying mechanism has not yet been fully clarified. We previously found that Sex-determining region of Y-related high-mobility-group box 6 (SOX6), a tumor suppressor gene or oncogene in several cancers, could induce autophagy in cervical cancer. Accordingly, this study aims to investigate the mechanism of SOX6-induced autophagy and its potential significance in the platinum-based chemotherapy of cervical cancer. Firstly, we found that SOX6 could promote autophagy in cervical cancer cells depending on its HMG domain. Mitogen-activated protein kinase kinase kinase kinase-4 (MAP4K4) gene was identified as the direct target gene of SOX6, which was transcriptionally upregulated by binding the HMG domain of SOX6 protein to its double-binding sites within MAP4K4 gene promoter. MAP4K4 mediated the SOX6-induced autophagy through inhibiting PI3K-Akt-mTOR pathway and activating MAPK/ERK pathway. Further, the sensitivity of cervical cancer cells to cisplatin chemotherapy could be reduced by the SOX6-induced autophagy in vitro and in vivo, while such a phenomenon could be turned over by autophagy-specific inhibitor and MAP4K4 inhibitor, respectively. Moreover, cisplatin itself could promote the expression of endogenous SOX6 and subsequently the MAP4K4-mediated autophagy in cervical cancer cells, which might in turn reduce the sensitivity of these cells to cisplatin treatment. These findings uncovered the underlying mechanism and potential significance of SOX6-induced autophagy, and shed new light on the usage of MAP4K4 inhibitor or autophagy-specific inhibitor for sensitizing cervical cancer cells to the platinum-based chemotherapy.
    DOI:  https://doi.org/10.1038/s41419-021-04474-1
  46. Biomolecules. 2021 Dec 14. pii: 1871. [Epub ahead of print]11(12):
    The MAP Kinase Phosphatase MKP-1 Modulates Neurogenesis via Effects on BNIP3 and Autophagy.
    Yinghui Li, Marc W Halterman.
      Inherited and acquired defects in neurogenesis contribute to neurodevelopmental disorders, dysfunctional neural plasticity, and may underlie pathology in a range of neurodegenerative conditions. Mitogen-activated protein kinases (MAPKs) regulate the proliferation, survival, and differentiation of neural stem cells. While the balance between MAPKs and the family of MAPK dual-specificity phosphatases (DUSPs) regulates axon branching and synaptic plasticity, the specific role that DUSPs play in neurogenesis remains unexplored. In the current study, we asked whether the canonical DUSP, MAP Kinase Phosphatase-1 (MKP-1), influences neural stem cell differentiation and the extent to which DUSP-dependent autophagy is operational in this context. Under basal conditions, Mkp-1 knockout mice generated fewer doublecortin (DCX) positive neurons within the dentate gyrus (DG) characterized by the accumulation of LC3 puncta. Analyses of wild-type neural stem cell (NSC) differentiation in vitro revealed increased Mkp-1 mRNA expression during the initial 24-h period. Notably, Mkp-1 KO NSC differentiation produced fewer Tuj1-positive neurons and was associated with increased expression of the BCL2/adenovirus E1B 19-kD protein-interacting protein 3 (BNIP3) and levels of autophagy. Conversely, Bnip3 knockdown in differentiated Mkp-1 KO NSCs reduced levels of autophagy and increased neuronal yields. These results indicate that MKP-1 exerts a pro-neurogenic bias during a critical window in NSC differentiation by regulating BNIP3 and basal autophagy levels.
    Keywords:  BNIP3; MKP-1; autophagy; neurogenesis
    DOI:  https://doi.org/10.3390/biom11121871
  47. Hepatology. 2021 Dec 23.
    Autophagy Promotes Hepatic Cystogenesis in Polycystic Liver Disease via Depletion of Cholangiocyte Ciliogenic Proteins.
    Anatoliy I Masyuk, Tatyana V Masyuk, Christy E Trussoni, Nicholas E Pirius, Nicholas F LaRusso.
       BACKGROUNDS & AIMS: Polycystic liver disease (PLD) is characterized by defective cholangiocyte cilia that regulate progressive growth of hepatic cysts. Because formation of primary cilia is influenced by autophagy through degradation of proteins involved in ciliogenesis, we hypothesized that ciliary defects in PLD cholangiocytes (PLDC) originate from autophagy-mediated depletion of ciliogenic proteins, ARL3 and ARL13B, and ARL-dependent mislocation of a ciliary-localized bile acid receptor, TGR5, activation of which enhances hepatic cystogenesis. The aims here were to determine: i) if ciliogenesis is impaired in PLDC, is associated with increased autophagy, and involves autophagy-mediated depletion of ARL3 and ARL13B; ii) if depletion of ARL3 and ARL13B in PLDC cilia impacts ciliary localization of TGR5; and iii) if pharmacological inhibition of autophagy reestablishes cholangiocyte cilia, ciliary localization of ARL3, ARL3B and TGR5, and reduces hepatic cystogenesis.
    APPROACH & RESULTS: By using liver tissue from healthy individuals and patients with PLD, in vitro and in vivo models of PLD, and in vitro models of ciliogenesis, we demonstrated that in PLDC: ciliogenesis is impaired; autophagy is enhanced; ARL3 and ARL13B are ubiquitinated by HDAC6, depleted in cilia and present in autophagosomes; depletion of ARL3 and ARL13B impacts ciliary localization of TGR5; and pharmacologic inhibition of autophagy with mefloquine and verteporfin reestablishes cholangiocyte cilia, ciliary localization of ARL3, ARL13B and TGR5, and reduces hepatic cystogenesis.
    CONCLUSION: The intersection between autophagy, defective cholangiocyte cilia, and enhanced hepatic cystogenesis contributes to PLD progression and can be considered a novel target for therapeutic interventions.
    Keywords:  Polycystic liver disease; cholangiocytes; ciliogenesis; primary cilia
    DOI:  https://doi.org/10.1002/hep.32298
  48. Cell Death Dis. 2021 Dec 18. 13(1): 7
    Sidt2 is a key protein in the autophagy-lysosomal degradation pathway and is essential for the maintenance of kidney structure and filtration function.
    Meng-Ya Geng, Lizhuo Wang, Ying-Ying Song, Jing Gu, Xin Hu, Cheng Yuan, Meng Yang, Wen-Jun Pei, Yao Zhang, Jia-Lin Gao.
      The regulation and homeostasis of autophagy are essential for maintaining organ morphology and function. As a lysosomal membrane protein, the effect of Sidt2 on kidney structure and renal autophagy is still unknown. In this study, we found that the kidneys of Sidt2-/- mice showed changes in basement membrane thickening, foot process fusion, and mitochondrial swelling, suggesting that the structure of the kidney was damaged. Increased urine protein at 24 h indicated that the kidney function was also damaged. At the same time, the absence of Sidt2 caused a decrease in the number of acidic lysosomes, a decrease in acid hydrolase activity and expression in the lysosome, and an increase of pH in the lysosome, suggesting that lysosomal function was impaired after Sidt2 deletion. The accumulation of autophagolysosomes, increased LC3-II and P62 protein levels, and decreased P62 mRNA levels indicated that the absence of the Sidt2 gene caused abnormal autophagy pathway flow. Chloroquine experiment, immunofluorescence autophagosome, and lysosome fusion assay, and Ad-mcherry-GFP-LC3B further indicated that, after Sidt2 deletion, the production of autophagosomes did not increase, but the fusion of autophagosomes and lysosomes and the degradation of autophagolysosomes were impaired. When incubating Sidt2-/- cells with the autophagy activator rapamycin, we found that it could activate autophagy, which manifested as an increase in autophagosomes, but it could not improve autophagolysosome degradation. Meanwhile, it further illustrated that the Sidt2 gene plays an important role in the smooth progress of autophagolysosome processes. In summary, the absence of the Sidt2 gene caused impaired lysosome function and a decreased number of acidic lysosomes, leading to formation and degradation disorders of the autophagolysosomes, which eventually manifested as abnormal kidney structure and function. Sidt2 is essential in maintaining the normal function of the lysosomes and the physiological stability of the kidneys.
    DOI:  https://doi.org/10.1038/s41419-021-04453-6
  49. Redox Biol. 2021 Dec 10. pii: S2213-2317(21)00370-0. [Epub ahead of print]49 102210
    Implication of type 4 NADPH oxidase (NOX4) in tauopathy.
    Enrique Luengo, Paula Trigo-Alonso, Cristina Fernández-Mendívil, Ángel Nuñez, Marta Del Campo, César Porrero, Nuria García-Magro, Pilar Negredo, Sergio Senar, Cristina Sánchez-Ramos, Juan A Bernal, Alberto Rábano, Jeroen Hoozemans, Ana I Casas, Harald H H W Schmidt, Manuela G López.
      Aggregates of the microtubule-associated protein tau are a common marker of neurodegenerative diseases collectively termed as tauopathies, such as Alzheimer's disease (AD) and frontotemporal dementia. Therapeutic strategies based on tau have failed in late stage clinical trials, suggesting that tauopathy may be the consequence of upstream causal mechanisms. As increasing levels of reactive oxygen species (ROS) may trigger protein aggregation or modulate protein degradation and, we had previously shown that the ROS producing enzyme NADPH oxidase 4 (NOX4) is a major contributor to cellular autotoxicity, this study was designed to evaluate if NOX4 is implicated in tauopathy. Our results show that NOX4 is upregulated in patients with frontotemporal lobar degeneration and AD patients and, in a humanized mouse model of tauopathy induced by AVV-TauP301L brain delivery. Both, global knockout and neuronal knockdown of the Nox4 gene in mice, diminished the accumulation of pathological tau and positively modified established tauopathy by a mechanism that implicates modulation of the autophagy-lysosomal pathway (ALP) and, consequently, improving the macroautophagy flux. Moreover, neuronal-targeted NOX4 knockdown was sufficient to reduce neurotoxicity and prevent cognitive decline, even after induction of tauopathy, suggesting a direct and causal role for neuronal NOX4 in tauopathy. Thus, NOX4 is a previously unrecognized causative, mechanism-based target in tauopathies and blood-brain barrier permeable specific NOX4 inhibitors could have therapeutic potential even in established disease.
    Keywords:  Alzheimer's disease; Autophagy; NADPH oxidases; NOX4; Tauopathy
    DOI:  https://doi.org/10.1016/j.redox.2021.102210
  50. Int J Mol Sci. 2021 Dec 09. pii: 13256. [Epub ahead of print]22(24):
    Impaired Retromer Function in Niemann-Pick Type C Disease Is Dependent on Intracellular Cholesterol Accumulation.
    Kristina Dominko, Ana Rastija, Sandra Sobocanec, Lea Vidatic, Sarah Meglaj, Andrea Lovincic Babic, Birgit Hutter-Paier, Alessio-Vittorio Colombo, Stefan F Lichtenthaler, Sabina Tahirovic, Silva Hecimovic.
      Niemann-Pick type C disease (NPC) is a rare inherited neurodegenerative disorder characterized by an accumulation of intracellular cholesterol within late endosomes and lysosomes due to NPC1 or NPC2 dysfunction. In this work, we tested the hypothesis that retromer impairment may be involved in the pathogenesis of NPC and may contribute to increased amyloidogenic processing of APP and enhanced BACE1-mediated proteolysis observed in NPC disease. Using NPC1-null cells, primary mouse NPC1-deficient neurons and NPC1-deficient mice (BALB/cNctr-Npc1m1N), we show that retromer function is impaired in NPC. This is manifested by altered transport of the retromer core components Vps26, Vps35 and/or retromer receptor sorLA and by retromer accumulation in neuronal processes, such as within axonal swellings. Changes in retromer distribution in NPC1 mouse brains were observed already at the presymptomatic stage (at 4-weeks of age), indicating that the retromer defect occurs early in the course of NPC disease and may contribute to downstream pathological processes. Furthermore, we show that cholesterol depletion in NPC1-null cells and in NPC1 mouse brains reverts retromer dysfunction, suggesting that retromer impairment in NPC is mechanistically dependent on cholesterol accumulation. Thus, we characterized retromer dysfunction in NPC and propose that the rescue of retromer impairment may represent a novel therapeutic approach against NPC.
    Keywords:  NPC1; cholesterol homeostasis; endolysosomal pathway; neurodegeneration; neurodegenerative diseases; rare diseases; retromer
    DOI:  https://doi.org/10.3390/ijms222413256
  51. Antioxidants (Basel). 2021 Nov 25. pii: 1884. [Epub ahead of print]10(12):
    Autophagy Is Involved in the Viability of Overexpressing Thioredoxin o1 Tobacco BY-2 Cells under Oxidative Conditions.
    Sabrina De Brasi-Velasco, Omar López-Vidal, María Carmen Martí, Ana Ortiz-Espín, Francisca Sevilla, Ana Jiménez.
      Autophagy is an essential process for the degradation of non-useful components, although the mechanism involved in its regulation is less known in plants than in animal systems. Redox regulation of autophagy components is emerging as a possible key mechanism with thioredoxins (TRXs) proposed as involved candidates. In this work, using overexpressing PsTRXo1 tobacco cells (OEX), which present higher viability than non-overexpressing cells after H2O2 treatment, we examine the functional interaction of autophagy and PsTRXo1 in a collaborative response. OEX cells present higher gene expression of the ATG (Autophagy related) marker ATG4 and higher protein content of ATG4, ATG8, and lipidated ATG8 as well as higher ATG4 activity than control cells, supporting the involvement of autophagy in their response to H2O2. In this oxidative situation, autophagy occurs in OEX cells as is evident from an accumulation of autolysosomes and ATG8 immunolocalization when the E-64d autophagy inhibitor is used. Interestingly, cell viability decreases in the presence of the inhibitor, pointing to autophagy as being involved in cell survival. The in vitro interaction of ATG4 and PsTRXo1 proteins is confirmed by dot-blot and co-immunoprecipitation assays as well as the redox regulation of ATG4 activity by PsTRXo1. These findings extend the role of TRXs in mediating the redox regulation of the autophagy process in plant cells.
    Keywords:  ATG4; autophagy flux; cell death; hydrogen peroxide; protein interaction; redox regulation
    DOI:  https://doi.org/10.3390/antiox10121884
  52. Int J Mol Sci. 2021 Dec 16. pii: 13511. [Epub ahead of print]22(24):
    Interdependent Regulation of Polycystin Expression Influences Starvation-Induced Autophagy and Cell Death.
    Jean-Paul Decuypere, Dorien Van Giel, Peter Janssens, Ke Dong, Stefan Somlo, Yiqiang Cai, Djalila Mekahli, Rudi Vennekens.
      Autosomal dominant polycystic kidney disease (ADPKD) is mainly caused by deficiency of polycystin-1 (PC1) or polycystin-2 (PC2). Altered autophagy has recently been implicated in ADPKD progression, but its exact regulation by PC1 and PC2 remains unclear. We therefore investigated cell death and survival during nutritional stress in mouse inner medullary collecting duct cells (mIMCDs), either wild-type (WT) or lacking PC1 (PC1KO) or PC2 (PC2KO), and human urine-derived proximal tubular epithelial cells (PTEC) from early-stage ADPKD patients with PC1 mutations versus healthy individuals. Basal autophagy was enhanced in PC1-deficient cells. Similarly, following starvation, autophagy was enhanced and cell death reduced when PC1 was reduced. Autophagy inhibition reduced cell death resistance in PC1KO mIMCDs to the WT level, implying that PC1 promotes autophagic cell survival. Although PC2 expression was increased in PC1KO mIMCDs, PC2 knockdown did not result in reduced autophagy. PC2KO mIMCDs displayed lower basal autophagy, but more autophagy and less cell death following chronic starvation. This could be reversed by overexpression of PC1 in PC2KO. Together, these findings indicate that PC1 levels are partially coupled to PC2 expression, and determine the transition from renal cell survival to death, leading to enhanced survival of ADPKD cells during nutritional stress.
    Keywords:  ADPKD; autophagy; autosomal dominant polycystic kidney disease; cell death; nutrient stress; polycystins
    DOI:  https://doi.org/10.3390/ijms222413511
  53. Elife. 2021 Dec 23. pii: e72593. [Epub ahead of print]10
    Disruption of the TCA cycle reveals an ATF4-dependent integration of redox and amino acid metabolism.
    Dylan Gerard Ryan, Ming Yang, Hiran A Prag, Giovanny Rodriguez Blanco, Efterpi Nikitopoulou, Marc Segarra-Mondejar, Christopher A Powell, Tim Young, Nils Burger, Jan Lj Miljkovic, Michal Minczuk, Michael P Murphy, Alex von Kriegsheim, Christian Frezza.
      The Tricarboxylic Acid Cycle (TCA) cycle is arguably the most critical metabolic cycle in physiology and exists as an essential interface coordinating cellular metabolism, bioenergetics, and redox homeostasis. Despite decades of research, a comprehensive investigation into the consequences of TCA cycle dysfunction remains elusive. Here, we targeted two TCA cycle enzymes, fumarate hydratase (FH) and succinate dehydrogenase (SDH), and combined metabolomics, transcriptomics, and proteomics analyses to fully appraise the consequences of TCA cycle inhibition (TCAi) in murine kidney epithelial cells. Our comparative approach shows that TCAi elicits a convergent rewiring of redox and amino acid metabolism dependent on the activation of ATF4 and the integrated stress response (ISR). Furthermore, we also uncover a divergent metabolic response, whereby acute FHi, but not SDHi, can maintain asparagine levels via reductive carboxylation and maintenance of cytosolic aspartate synthesis. Our work highlights an important interplay between the TCA cycle, redox biology and amino acid homeostasis.
    Keywords:  biochemistry; cell biology; chemical biology; mouse
    DOI:  https://doi.org/10.7554/eLife.72593
  54. Free Radic Biol Med. 2021 Dec 18. pii: S0891-5849(21)01115-1. [Epub ahead of print]
    Polygala saponins inhibit NLRP3 inflammasome-mediated neuroinflammation via SHP2-Mediated mitophagy.
    Wen-Qiao Qiu, Wei Ai, Feng-Dan Zhu, Yue Zhang, Min-Song Guo, Betty Yuen-Kwan Law, Jian-Ming Wu, Vincent Kam-Wai Wong, Yong Tang, Lu Yu, Qi Chen, Chong-Lin Yu, Jian Liu, Da-Lian Qin, Xiao-Gang Zhou, An-Guo Wu.
      Activation of the NLRP3 inflammasome and its mediated neuroinflammation are implicated in neurodegenerative diseases, while mitophagy negatively regulates NLRP3 inflammasome activation. SHP-2, a protein-tyrosine phosphatase, is critical for NLRP3 inflammasome regulation and inflammatory responses. In this study, we investigated whether triterpenoid saponins in Radix Polygalae inhibit the NLRP3 inflammasome via mitophagy induction. First, we isolated the active fraction (polygala saponins (PSS)) and identified 17 saponins by ultra-performance liquid chromatography coupled with diode-array detection and tandem quadrupole time-of-flight mass spectrometry (UHPLC-DAD-Q/TOF-MS). In microglial BV-2 cells, PSS induced mitophagy as evidenced by increased co-localization of LC3 and mitochondria, as well as an increased number of autophagic vacuoles surrounding the mitochondria. Furthermore, the mechanistic study found that PSS activated the AMPK/mTOR and PINK1/parkin signaling pathways via the upregulation of SHP-2. In Aβ(1-42)-, A53T-α-synuclein-, or Q74-induced BV-2 cells, PSS significantly inhibited NLRP3 inflammasome activation, which was attenuated by bafilomycin A1 (an autophagy inhibitor) and SHP099 (an SHP-2 inhibitor). In addition, the co-localization of LC3 and ASC revealed that PSS promoted the autophagic degradation of the NLRP3 inflammasome. Moreover, PSS decreased apoptosis in conditioned medium-induced PC-12 cells. In APP/PS1 mice, PSS improved cognitive function, ameliorated Aβ pathology, and inhibited neuronal death. Collectively, the present study, for the first time, shows that PSS inhibit the NLRP3 inflammasome via SHP-2-mediated mitophagy in vitro and in vivo, which strongly suggests the therapeutic potential of PSS in various neurodegenerative diseases.
    Keywords:  Mitophagy; NLRP3 inflammasome; Neurodegenerative disease; Polygala saponins; Radix polygalae; SHP-2
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2021.12.263
  55. Antioxidants (Basel). 2021 Dec 03. pii: 1944. [Epub ahead of print]10(12):
    mTOR Inhibition via Rapamycin Treatment Partially Reverts the Deficit in Energy Metabolism Caused by FH Loss in RPE Cells.
    David A Merle, Francesca Provenzano, Mohamed Ali Jarboui, Ellen Kilger, Simon J Clark, Michela Deleidi, Angela Armento, Marius Ueffing.
      Age-related macular degeneration (AMD) is a complex degenerative disease of the retina with multiple risk-modifying factors, including aging, genetics, and lifestyle choices. The combination of these factors leads to oxidative stress, inflammation, and metabolic failure in the retinal pigment epithelium (RPE) with subsequent degeneration of photoreceptors in the retina. The alternative complement pathway is tightly linked to AMD. In particular, the genetic variant in the complement factor H gene (CFH), which leads to the Y402H polymorphism in the factor H protein (FH), confers the second highest risk for the development and progression of AMD. Although the association between the FH Y402H variant and increased complement system activation is known, recent studies have uncovered novel FH functions not tied to this activity and highlighted functional relevance for intracellular FH. In our previous studies, we show that loss of CFH expression in RPE cells causes profound disturbances in cellular metabolism, increases the vulnerability towards oxidative stress, and modulates the activation of pro-inflammatory signaling pathways, most importantly the NF-kB pathway. Here, we silenced CFH in hTERT-RPE1 cells to investigate the mechanism by which intracellular FH regulates RPE cell homeostasis. We found that silencing of CFH results in hyperactivation of mTOR signaling along with decreased mitochondrial respiration and that mTOR inhibition via rapamycin can partially rescue these metabolic defects. To obtain mechanistic insight into the function of intracellular FH in hTERT-RPE1 cells, we analyzed the interactome of FH via immunoprecipitation followed by mass spectrometry-based analysis. We found that FH interacts with essential components of the ubiquitin-proteasomal pathway (UPS) as well as with factors associated with RB1/E2F signalling in a complement-pathway independent manner. Moreover, we found that FH silencing affects mRNA levels of the E3 Ubiquitin-Protein Ligase Parkin and PTEN induced putative kinase (Pink1), both of which are associated with UPS. As inhibition of mTORC1 was previously shown to result in increased overall protein degradation via UPS and as FH mRNA and protein levels were shown to be affected by inhibition of UPS, our data stress a potential regulatory link between endogenous FH activity and the UPS.
    Keywords:  age-related macular degeneration (AMD); complement factor H (CFH/FH); interactome; mammalian target of rapamycin (mTOR); mitochondrial respiration; retinal pigment epithelium (RPE) cells; ubiquitin-proteasomal pathway
    DOI:  https://doi.org/10.3390/antiox10121944
  56. Biomolecules. 2021 Nov 30. pii: 1789. [Epub ahead of print]11(12):
    Advances in Proteasome Enhancement by Small Molecules.
    Dare E George, Jetze J Tepe.
      The proteasome system is a large and complex molecular machinery responsible for the degradation of misfolded, damaged, and redundant cellular proteins. When proteasome function is impaired, unwanted proteins accumulate, which can lead to several diseases including age-related and neurodegenerative diseases. Enhancing proteasome-mediated substrate degradation with small molecules may therefore be a valuable strategy for the treatment of various neurodegenerative diseases such as Parkinson's, Alzheimer's, and Huntington's diseases. In this review, we discuss the structure of proteasome and how proteasome's proteolytic activity is associated with aging and various neurodegenerative diseases. We also summarize various classes of compounds that are capable of enhancing, directly or indirectly, proteasome-mediated protein degradation.
    Keywords:  20S; 26S; cancer; degradation; disordered; misfolded; neurodegeneration; proteasome; protein; ubiquitin
    DOI:  https://doi.org/10.3390/biom11121789
  57. J Exp Med. 2022 Feb 07. pii: e20211523. [Epub ahead of print]219(2):
    Targeting PLD2 in adipocytes augments adaptive thermogenesis by improving mitochondrial quality and quantity in mice.
    Hyung Sik Kim, Min Young Park, Nam Joo Yun, Hye Sun Go, Mi Young Kim, Je Kyung Seong, Minyoung Lee, Eun Seok Kang, Jaewang Ghim, Sung Ho Ryu, Brian A Zabel, Ara Koh, Yoe-Sik Bae.
      Phospholipase D (PLD)2 via its enzymatic activity regulates cell proliferation and migration and thus is implicated in cancer. However, the role of PLD2 in obesity and type 2 diabetes has not previously been investigated. Here, we show that during diet-induced thermogenesis and obesity, levels of PLD2 but not PLD1 in adipose tissue are inversely related with uncoupling protein 1, a key thermogenic protein. We demonstrate that the thermogenic program in adipose tissue is significantly augmented in mice with adipocyte-specific Pld2 deletion or treated with a PLD2-specific inhibitor and that these mice are resistant to high fat diet-induced obesity, glucose intolerance, and insulin resistance. Mechanistically, we show that Pld2 deletion in adipose tissue or PLD2 pharmacoinhibition acts via p62 to improve mitochondrial quality and quantity in adipocytes. Thus, PLD2 inhibition is an attractive therapeutic approach for obesity and type 2 diabetes by resolving defects in diet-induced thermogenesis.
    DOI:  https://doi.org/10.1084/jem.20211523
  58. Toxins (Basel). 2021 Dec 18. pii: 909. [Epub ahead of print]13(12):
    Indoxyl Sulfate Contributes to mTORC1-Induced Renal Fibrosis via The OAT/NADPH Oxidase/ROS Pathway.
    Takehiro Nakano, Hiroshi Watanabe, Tadashi Imafuku, Kai Tokumaru, Issei Fujita, Nanaka Arimura, Hitoshi Maeda, Motoko Tanaka, Kazutaka Matsushita, Masafumi Fukagawa, Toru Maruyama.
      Activation of mTORC1 (mechanistic target of rapamycin complex 1) in renal tissue has been reported in chronic kidney disease (CKD)-induced renal fibrosis. However, the molecular mechanisms responsible for activating mTORC1 in CKD pathology are not well understood. The purpose of this study was to identify the uremic toxin involved in mTORC1-induced renal fibrosis. Among the seven protein-bound uremic toxins, only indoxyl sulfate (IS) caused significant activation of mTORC1 in human kidney 2 cells (HK-2 cells). This IS-induced mTORC1 activation was inhibited in the presence of an organic anion transporter inhibitor, a NADPH oxidase inhibitor, and an antioxidant. IS also induced epithelial-mesenchymal transition of tubular epithelial cells (HK-2 cells), differentiation of fibroblasts into myofibroblasts (NRK-49F cells), and inflammatory response of macrophages (THP-1 cells), which are associated with renal fibrosis, and these effects were inhibited in the presence of rapamycin (mTORC1 inhibitor). In in vivo experiments, IS overload was found to activate mTORC1 in the mouse kidney. The administration of AST-120 or rapamycin targeted to IS or mTORC1 ameliorated renal fibrosis in Adenine-induced CKD mice. The findings reported herein indicate that IS activates mTORC1, which then contributes to renal fibrosis. Therapeutic interventions targeting IS and mTORC1 could be effective against renal fibrosis in CKD.
    Keywords:  AST-120; chronic kidney disease; indoxyl sulfate; mTORC1; renal fibrosis
    DOI:  https://doi.org/10.3390/toxins13120909
  59. J Nanobiotechnology. 2021 Dec 24. 19(1): 447
    Biomimetic nanoparticles blocking autophagy for enhanced chemotherapy and metastasis inhibition via reversing focal adhesion disassembly.
    Yesi Shi, Gan Lin, Huili Zheng, Dan Mu, Hu Chen, Zhixiang Lu, Pan He, Yang Zhang, Chao Liu, Zhongning Lin, Gang Liu.
       BACKGROUND: Autophagy is a conserved catabolic process, which plays an important role in regulating tumor cell motility and degrading protein aggregates. Chemotherapy-induced autophagy may lead to tumor distant metastasis and even chemo-insensitivity in the therapy of hepatocellular carcinoma (HCC). Therefore, a vast majority of HCC cases do not produce a significant response to monotherapy with autophagy inhibitors.
    RESULTS: In this work, we developed a biomimetic nanoformulation (TH-NP) co-encapsulating Oxaliplatin (OXA)/hydroxychloroquine (HCQ, an autophagy inhibitor) to execute targeted autophagy inhibition, reduce tumor cell migration and invasion in vitro and attenuate metastasis in vivo. The tumor cell-specific ligand TRAIL was bioengineered to be stably expressed on HUVECs and the resultant membrane vesicles were wrapped on OXA/HCQ-loaded PLGA nanocores. Especially, TH-NPs could significantly improve OXA and HCQ effective concentration by approximately 21 and 13 times in tumor tissues compared to the free mixture of HCQ/OXA. Moreover, the tumor-targeting TH-NPs released HCQ alkalized the acidic lysosomes and inhibited the fusion of autophagosomes and lysosomes, leading to effective blockade of autophagic flux. In short, the system largely improved chemotherapeutic performance of OXA on subcutaneous and orthotopic HCC mice models. Importantly, TH-NPs also exhibited the most effective inhibition of tumor metastasis in orthotopic HCCLM3 models, and in the HepG2, Huh-7 or HCCLM3 metastatic mice models. Finally, we illustrated the enhanced metastasis inhibition was attributed to the blockade or reverse of the autophagy-mediated degradation of focal adhesions (FAs) including E-cadherin and paxillin.
    CONCLUSIONS: TH-NPs can perform an enhanced chemotherapy and antimetastatic effect, and may represent a promising strategy for HCC therapy in clinics.
    Keywords:  Autophagy inhibition; Biomimetic; Focal adhesions; Metastasis; Targeted co-delivery
    DOI:  https://doi.org/10.1186/s12951-021-01189-5
  60. Trends Endocrinol Metab. 2021 Dec 16. pii: S1043-2760(21)00267-8. [Epub ahead of print]
    Amino acid transporters as modulators of glucose homeostasis.
    Stefan Bröer.
      Amino acids modulate glucose homeostasis. Cytosolic levels of amino acids are regulated by amino acid transporters, modulating insulin release, protein synthesis, cell proliferation, cell fate, and metabolism. In β-cells, amino acid transporters modulate incretin-stimulated insulin release. In the liver, amino acid transporters provide glutamine and alanine for gluconeogenesis. Intestinal amino acid transporters facilitate the intake of amino acids causing protein restriction when inactive. Adipocyte development is regulated by amino acid transporters through activation of mechanistic target of rapamycin (mTORC1) and amino acid-related metabolites. The accumulation and metabolism of branched-chain amino acids (BCAAs) in muscle depends on transporters. The integration between amino acid metabolism and transport is critical for the maintenance and function of tissues and cells involved in glucose homeostasis.
    Keywords:  GCN2; adipocytes; gluconeogenesis; solute carrier; β-cell
    DOI:  https://doi.org/10.1016/j.tem.2021.11.004
  61. Front Cardiovasc Med. 2021 ;8 757591
    An Inhibitor of Grp94 Inhibits OxLDL-Induced Autophagy and Apoptosis in VECs and Stabilized Atherosclerotic Plaques.
    Qun Wei, Hui Ren, Jun Zhang, Wen Yao, Baoxiang Zhao, Junying Miao.
      Background: Oxidized low-density lipoprotein (oxLDL) induces vascular endothelial cell (VEC) injury and atherosclerosis through activating endoplasmic reticulum stress. Expression of glucose-regulated protein 94 (Grp94) is induced by endoplasmic reticulum stress and Grp94 is involved in cardiovascular diseases. This study aimed to determine the role of Grp94 in oxLDL-induced vascular endothelial cell injury and atherosclerosis. Methods and Results: An inhibitor of Grp94, HCP1, was used to investigate the role of Grp94 in oxLDL-induced VEC injury in human umbilical vein endothelial cells and atherosclerosis in apolipoprotein E-/- mice. Results showed that HCP1 inhibited autophagy and apoptosis induced by oxLDL in VECs. And we found that Grp94 might interact with adenosine monophosphate-activated protein kinase (AMPK) and activate its activity. HCP1 inhibited AMPK activity and overexpression of Grp94 blocked the effect of HCP1. Besides, HCP1 activated the activity of mechanistic target of rapamycin complex 1 (mTORC1), co-treatment with AMPK activator acadesine eliminated the effect of HCP1 on mTORC1 activity as well as autophagy. In apolipoprotein E-/- mice, HCP1 suppressed autophagy and apoptosis of atherosclerotic plaque endothelium. In addition, HCP1 increased the content of collagen, smooth muscle cells, and anti-inflammatory macrophages while reducing the activity of MMP-2/9 and pro-inflammatory macrophages in the atherosclerotic lesion. Conclusion: HCP1 inhibited oxLDL-induced VEC injury and promoted the stabilization of atherosclerotic plaque in apoE-/- mice. Grp94 might be a potential therapeutic target in the clinical treatment of atherosclerosis.
    Keywords:  adenosine monophosphate-activated protein kinase; apoptosis; atherosclerosis; autophagy; glucose-regulated protein 94
    DOI:  https://doi.org/10.3389/fcvm.2021.757591
  62. Int J Mol Sci. 2021 Dec 09. pii: 13271. [Epub ahead of print]22(24):
    Sequestosome 1 Is Part of the Interaction Network of VAPB.
    Christina James, Christof Lenz, Henning Urlaub, Ralph H Kehlenbach.
      VAPB (Vesicle-Associated-membrane Protein-associated protein B) is a tail-anchored membrane protein of the endoplasmic reticulum that can also be detected at the inner nuclear membrane. As a component of many contact sites between the endoplasmic reticulum and other organelles, VAPB is engaged in multiple protein interactions with a plethora of binding partners. A mutant version of VAPB, P56S-VAPB, which results from a single point mutation, is involved in a familial form of amyotrophic lateral sclerosis (ALS8). We performed RAPIDS (rapamycin- and APEX-dependent identification of proteins by SILAC) to identify proteins that interact with or are in close proximity to P56S-VAPB. The mutation abrogates the interaction of VAPB with many known binding partners. Here, we identify Sequestosome 1 (SQSTM1), a well-known autophagic adapter protein, as a major interaction/proximity partner of P56S-VAPB. Remarkably, not only the mutant protein, but also wild-type VAPB interacts with SQSTM1, as shown by proximity ligation assays and co-immunoprecipiation experiments.
    Keywords:  SQSTM1; VAPB; amyotrophic lateral sclerosis; p62; sequestosome
    DOI:  https://doi.org/10.3390/ijms222413271
  63. Biomedicines. 2021 Nov 27. pii: 1782. [Epub ahead of print]9(12):
    Metformin Improves Stemness of Human Adipose-Derived Stem Cells by Downmodulation of Mechanistic Target of Rapamycin (mTOR) and Extracellular Signal-Regulated Kinase (ERK) Signaling.
    Somaiah Chinnapaka, Katherine S Yang, Quinn Flowers, Minhal Faisal, Wayne Vincent Nerone, Joseph Peter Rubin, Asim Ejaz.
      Adipose tissue plays an important role in regulating metabolic homeostasis by storing excess fat and protecting other organs from lipotoxicity. Aging is associated with central fat redistribution, culminating in a decrease in insulin-sensitive subcutaneous and an increase in insulin-resistant visceral adipose depots. Adipose-derived stem cells (ASCs) play an important role in the regeneration of adipose tissue. Aged ASCs show decreased stemness and regenerative potential due to the accumulation of oxidative stress and mitochondrial dysfunction-related cell damage. Metformin is a well-established anti-diabetic drug that has shown anti-aging effects in different organisms and animal models. In this study, we analyzed the effect of metformin treatment on the stemness of human ASCs in cell culture and whole adipose tissue culture models. Our results demonstrate that metformin improves the stemness of ASCs, reducing their rate of proliferation and adipocyte differentiation. Investigating the possible underlying mechanism, we observed a decrease in the mTOR and ERK activity in metformin-treated ASCs. In addition, we observed an increase in autophagy activity upon metformin treatment. We conclude that metformin treatment improves ASCs stemness by reducing mTOR and ERK signaling and enhancing autophagy. Future in vivo evaluations in animal models and humans will pave the way for the clinical adaptation of this well-established drug for reviving the stemness of aged stem cells.
    Keywords:  ERK; adipose stem cells; adipose tissue; autophagy; differentiation; mTOR; metformin; proliferation; stemness
    DOI:  https://doi.org/10.3390/biomedicines9121782
  64. Biomolecules. 2021 Nov 26. pii: 1775. [Epub ahead of print]11(12):
    Therapeutic Approaches in Lysosomal Storage Diseases.
    Carlos Fernández-Pereira, Beatriz San Millán-Tejado, María Gallardo-Gómez, Tania Pérez-Márquez, Marta Alves-Villar, Cristina Melcón-Crespo, Julián Fernández-Martín, Saida Ortolano.
      Lysosomal Storage Diseases are multisystemic disorders determined by genetic variants, which affect the proteins involved in lysosomal function and cellular metabolism. Different therapeutic approaches, which are based on the physiologic mechanisms that regulate lysosomal function, have been proposed for these diseases. Currently, enzyme replacement therapy, gene therapy, or small molecules have been approved or are under clinical development to treat lysosomal storage disorders. The present article reviews the main therapeutic strategies that have been proposed so far, highlighting possible limitations and future perspectives.
    Keywords:  autophagy; enzyme replacement therapy; gene therapy; lysosomal storage diseases; small molecules
    DOI:  https://doi.org/10.3390/biom11121775
  65. Cancers (Basel). 2021 Dec 10. pii: 6220. [Epub ahead of print]13(24):
    Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators.
    David P Dannheisig, Anna Schimansky, Cornelia Donow, Astrid S Pfister.
      Ribosome biogenesis is essential for protein synthesis, cell growth and survival. The process takes places in nucleoli and is orchestrated by various proteins, among them RNA polymerases I-III as well as ribosome biogenesis factors. Perturbation of ribosome biogenesis activates the nucleolar stress response, which classically triggers cell cycle arrest and apoptosis. Nucleolar stress is utilized in modern anti-cancer therapies, however, also contributes to the development of various pathologies, including cancer. Growing evidence suggests that nucleolar stress stimulates compensatory cascades, for instance bulk autophagy. However, underlying mechanisms are poorly understood. Here we demonstrate that induction of nucleolar stress activates expression of key autophagic regulators such as ATG7 and ATG16L1, essential for generation of autophagosomes. We show that knockdown of the ribosomopathy factor SBDS, or of key ribosome biogenesis factors (PPAN, NPM, PES1) is associated with enhanced levels of ATG7 in cancer cells. The same holds true when interfering with RNA polymerase I function by either pharmacological inhibition (CX-5461) or depletion of the transcription factor UBF-1. Moreover, we demonstrate that RNA pol I inhibition by CX-5461 stimulates autophagic flux. Together, our data establish that nucleolar stress affects transcriptional regulation of autophagy. Given the contribution of both axes in propagation or cure of cancer, our data uncover a connection that might be targeted in future.
    Keywords:  ATG16L1; ATG7; CX-5461; NPM; PES1; PPAN; SBDS; UBF-1; autophagy; nucleolar stress; nucleolus; ribosome biogenesis
    DOI:  https://doi.org/10.3390/cancers13246220
  66. Antioxidants (Basel). 2021 Nov 29. pii: 1916. [Epub ahead of print]10(12):
    Camphorquinone Promotes the Antisenescence Effect via Activating AMPK/SIRT1 in Stem Cells and D-Galactose-Induced Aging Mice.
    Nagarajan Maharajan, Gwang-Won Cho.
      Terpenoids are a wide class of secondary metabolites with geroprotective properties that can alter the mechanism of aging and aging-related diseases. Camphorquinone (CQ) is a bicyclic monoterpenoid compound that can be efficiently synthesized through the continuous bromination and oxidation reaction of camphor. The purpose of this study is to investigate the effects of CQ on oxidative-stress-induced senescence and its underlying mechanisms. To generate oxidative stress in human bone marrow mesenchymal stem cells (hBM-MSCs) and mice, we used hydrogen peroxide (200 μM twice) and D-galactose (D-Gal) (150 mg/kg for 10 weeks), respectively. Our findings suggest that CQ potentially reduces senescence in hBM-MSCs and mouse heart tissue. In addition, we found that CQ boosted AMPK/SIRT1 activation and autophagy in both models. These results were subsequently verified in hBM-MSCs using compound C (an AMPK inhibitor) but AMPK inhibition by CC did not significantly reduce the SIRT1 and the autophagy markers. CQ treatment also reduced the gene expression of inflammation markers in D-Gal-induced aging mouse heart tissue. Furthermore, we determined that CQ fits all of the pharmacological parameters using the freely available SwissADME Web tool. Collectively, our findings demonstrate that CQ possesses antisenescence and cardioprotective properties, and that oxidative-stress-induced senescence could be suppressed by AMPK/SIRT1 and autophagy mechanisms.
    Keywords:  AMPK; SIRT1; autophagy; camphorquinone; oxidative stress; senescence
    DOI:  https://doi.org/10.3390/antiox10121916
  67. Cancers (Basel). 2021 Dec 14. pii: 6280. [Epub ahead of print]13(24):
    HDAC6 Inhibition Extinguishes Autophagy in Cancer: Recent Insights.
    Eugenia Passaro, Chiara Papulino, Ugo Chianese, Antonella Toraldo, Raffaella Congi, Nunzio Del Gaudio, Maria Maddalena Nicoletti, Rosaria Benedetti, Lucia Altucci.
      Autophagy is an essential intracellular catabolic mechanism involved in the degradation and recycling of damaged organelles regulating cellular homeostasis and energy metabolism. Its activation enhances cellular tolerance to various stresses and is known to be involved in drug resistance. In cancer, autophagy has a dual role in either promoting or blocking tumorigenesis, and recent studies indicate that epigenetic regulation is involved in its mechanism of action in this context. Specifically, the ubiquitin-binding histone deacetylase (HDAC) enzyme HDAC6 is known to be an important player in modulating autophagy. Epigenetic modulators, such as HDAC inhibitors, mediate this process in different ways and are already undergoing clinical trials. In this review, we describe current knowledge on the role of epigenetic modifications, particularly HDAC-mediated modifications, in controlling autophagy in cancer. We focus on the controversy surrounding their ability to promote or block tumor progression and explore the impact of HDAC6 inhibitors on autophagy modulation in cancer. In light of the fact that targeted drug therapy for cancer patients is attracting ever increasing interest within the research community and in society at large, we discuss the possibility of using HDAC6 inhibitors as adjuvants and/or in combination with conventional treatments to overcome autophagy-related mechanisms of resistance.
    Keywords:  HDAC inhibitors; HDAC6; autophagy; cancer; drug resistance; epigenetics
    DOI:  https://doi.org/10.3390/cancers13246280
  68. Biomed Res. 2021 ;42(6): 239-246
    Long non-coding RNA TUG1 promotes the osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the AMPK/mTOR/autophagy pathway.
    Ding-Gui Lu, Mei-Jiao Lu, Shun-Han Yao, Jia-Jie Lin, Su Luo, Ji-Hua Wei, Yu-Jin Tang.
      Promoting the differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts is an effective strategy against osteoporosis. Long non-coding RNAs are closely implicated in BMSC osteogenic differentiation. The present study explored the expression pattern and biological role of taurine upregulated gene 1 (TUG1) in osteogenic differentiation. The expressions of TUG1 and osteogenic markers following the osteogenic induction of BMSCs were detected. The functional relevance of TUG1 was evaluated by performing gain- and loss-of-function tests. Inhibitors of AMP-activated protein kinase (AMPK) autophagy were applied to ascertain the effects of TUG1 on the osteogenic differentiation of BMSCs. TUG1 expression increased during the osteogenic differentiation of BMSCs. The overexpression of TUG1 was promoted, whereas the knockdown of TUG1 was suppressed, by BMSC osteogenic differentiation. Mechanically, TUG1 promoted the osteogenesis of BMSCs via the AMPK-mammalian target of rapamycin (mTOR)-autophagy signaling pathway. Blocking AMPK and autophagy could abrogate the osteogenic role of TUG1 in BMSCs. These results demonstrated that TUG1 promoted the osteogenic differentiation of BMSCs by regulating the AMPK/mTOR/autophagy axis, suggesting that targeting TUG1 may be a potential therapy for osteoporosis.
    DOI:  https://doi.org/10.2220/biomedres.42.239
  69. Front Cell Dev Biol. 2021 ;9 748485
    Crosstalk Between Autophagy and the cGAS-STING Signaling Pathway in Type I Interferon Production.
    Kunli Zhang, Sutian Wang, Hongchao Gou, Jianfeng Zhang, Chunling Li.
      Innate immunity is the front-line defense against infectious microorganisms, including viruses and bacteria. Type I interferons are pleiotropic cytokines that perform antiviral, antiproliferative, and immunomodulatory functions in cells. The cGAS-STING pathway, comprising the main DNA sensor cyclic guanosine monophosphate/adenosine monophosphate synthase (cGAS) and stimulator of IFN genes (STING), is a major pathway that mediates immune reactions and is involved in the strong induction of type I IFN production, which can fight against microbial infections. Autophagy is an evolutionarily conserved degradation process that is required to maintain host health and facilitate capture and elimination of invading pathogens by the immune system. Mounting evidence indicates that autophagy plays an important role in cGAS-STING signaling pathway-mediated type I IFN production. This review briefly summarizes the research progress on how autophagy regulates the cGAS-STING pathway, regulating type I IFN production, with a particular focus on the crosstalk between autophagy and cGAS-STING signaling during infection by pathogenic microorganisms.
    Keywords:  CGAS; STING; autophagy; signalling pathway; type I IFNs production
    DOI:  https://doi.org/10.3389/fcell.2021.748485
  70. Autophagy. 2021 Dec 22. 1-14
    SQSTM1-mediated clearance of cytoplasmic mutant TARDBP/TDP-43 in the monkey brain.
    Peng Yin, Dazhang Bai, Fuyu Deng, Chen Zhang, Qingqing Jia, Longhong Zhu, Laiqiang Chen, Bang Li, Xiangyu Guo, Jianmeng Ye, Zhiqiang Tan, Lu Wang, Shihua Li, Xiao-Jiang Li.
      The cytoplasmic accumulation and aggregates of TARDBP/TDP-43 (TAR DNA binding protein) are a pathological hallmark in amyotrophic lateral sclerosis and frontotemporal lobar degeneration. We previously reported that the primate specific cleavage of TARDBP accounts for its cytoplasmic mislocalization in the primate brains, prompting us to further investigate how the cytoplasmic TARDBP mediates neuropathology. Here we reported that cytoplasmic mutant TARDBP reduced SQSTM1 expression selectively in the monkey brain, when compared with the mouse brain, by inducing SQSTM1 mRNA instability via its binding to the unique 3'UTR sequence (GU/UG)n of the primate SQSTM1 transcript. Overexpression of SQSTM1 could diminish the cytoplasmic C-terminal TARDBP accumulation in the monkey brain by augmenting macroautophagy/autophagy activity. Our findings provide additional clues for the pathogenesis of cytoplasmic TARDBP and a potential therapy for mutant TARDBP-mediated neuropathology.
    Keywords:  Aggregate; RNA instability; SQSTM1; TARDBP; non-human primate
    DOI:  https://doi.org/10.1080/15548627.2021.2013653
  71. Antioxidants (Basel). 2021 Nov 30. pii: 1924. [Epub ahead of print]10(12):
    Resolvin D1 Suppresses H2O2-Induced Senescence in Fibroblasts by Inducing Autophagy through the miR-1299/ARG2/ARL1 Axis.
    Hyun Ji Kim, Boram Kim, Hyung Jung Byun, Lu Yu, Tuan Minh Nguyen, Thi Ha Nguyen, Phuong Anh Do, Eun Ji Kim, Kyung Ah Cheong, Kyung Sung Kim, Hiệu Huy Phùng, Mostafizur Rahman, Ji Yun Jang, Seung Bae Rho, Gyeoung Jin Kang, Mi Kyung Park, Ho Lee, Kyeong Lee, Jungsook Cho, Hyo Kyung Han, Sang Geon Kim, Ai Young Lee, Chang Hoon Lee.
      ARG2 has been reported to inhibit autophagy in vascular endothelial cells and keratinocytes. However, studies of its mechanism of action, its role in skin fibroblasts, and the possibility of promoting autophagy and inhibiting cellular senescence through ARG2 inhibition are lacking. We induced cellular senescence in dermal fibroblasts by using H2O2. H2O2-induced fibroblast senescence was inhibited upon ARG2 knockdown and promoted upon ARG2 overexpression. The microRNA miR-1299 suppressed ARG2 expression, thereby inhibiting fibroblast senescence, and miR-1299 inhibitors promoted dermal fibroblast senescence by upregulating ARG2. Using yeast two-hybrid assay, we found that ARG2 binds to ARL1. ARL1 knockdown inhibited autophagy and ARL1 overexpression promoted it. Resolvin D1 (RvD1) suppressed ARG2 expression and cellular senescence. These data indicate that ARG2 stimulates dermal fibroblast cell senescence by inhibiting autophagy after interacting with ARL1. In addition, RvD1 appears to promote autophagy and inhibit dermal fibroblast senescence by inhibiting ARG2 expression. Taken together, the miR-1299/ARG2/ARL1 axis emerges as a novel mechanism of the ARG2-induced inhibition of autophagy. Furthermore, these results indicate that miR-1299 and pro-resolving lipids, including RvD1, are likely involved in inhibiting cellular senescence by inducing autophagy.
    Keywords:  ARG2; ARL1; H2O2; autophagy; cell senescence; fibroblast; miR-1299
    DOI:  https://doi.org/10.3390/antiox10121924
  72. Nat Immunol. 2021 Dec 23.
    The kinase complex mTORC2 promotes the longevity of virus-specific memory CD4+ T cells by preventing ferroptosis.
    Yifei Wang, Qin Tian, Yaxing Hao, Wei Yao, Jinjin Lu, Cheng Chen, Xiangyu Chen, Yao Lin, Qizhao Huang, Lifan Xu, Jianjun Hu, Shun Lei, Zhengping Wei, Yuan Luo, Zhirong Li, Li Hu, Jianfang Tang, Qing Wu, Xinyuan Zhou, Yuzhang Wu, Zhinan Yin, Jianqing Xu, Lilin Ye.
      Antigen-specific memory CD4+ T cells can persist and confer rapid and efficient protection from microbial reinfection. However, the mechanisms underlying the long-term maintenance of the memory CD4+ T cell pool remain largely unknown. Here, using a mouse model of acute infection with lymphocytic choriomeningitis virus (LCMV), we found that the serine/threonine kinase complex mammalian target of rapamycin complex 2 (mTORC2) is critical for the long-term persistence of virus-specific memory CD4+ T cells. The perturbation of mTORC2 signaling at memory phase led to an enormous loss of virus-specific memory CD4+ T cells by a unique form of regulated cell death (RCD), ferroptosis. Mechanistically, mTORC2 inactivation resulted in the impaired phosphorylation of downstream AKT and GSK3β kinases, which induced aberrant mitochondrial reactive oxygen species (ROS) accumulation and ensuing ferroptosis-causative lipid peroxidation in virus-specific memory CD4+ T cells; furthermore, the disruption of this signaling cascade also inhibited glutathione peroxidase 4 (GPX4), a major scavenger of lipid peroxidation. Thus, the mTORC2-AKT-GSK3β axis functions as a key signaling hub to promote the longevity of virus-specific memory CD4+ T cells by preventing ferroptosis.
    DOI:  https://doi.org/10.1038/s41590-021-01090-1
  73. Brain. 2021 Dec 20. pii: awab467. [Epub ahead of print]
    Early lysosome defects precede neurodegeneration with amyloid-β and tau aggregation in NHE6-null rat brain.
    YouJin Lee, Morgan R Miller, Marty A Fernandez, Elizabeth L Berg, Adriana M Prada, Qing Ouyang, Michael Schmidt, Jill L Silverman, Tracy L Young-Pearse, Eric M Morrow.
      Loss-of-function mutations in the X-linked endosomal Na+/H+ Exchanger 6 (NHE6) cause Christianson syndrome (CS) in males. CS involves endosome dysfunction leading to early cerebellar degeneration, as well as later-onset cortical and subcortical neurodegeneration, potentially including tau deposition as reported in postmortem studies. In addition, there is reported evidence of modulation of amyloid beta (Aβ) levels in experimental models wherein NHE6 expression was targeted. We have recently shown that loss of NHE6 causes defects in endosome maturation and trafficking underlying lysosome deficiency in primary mouse neurons in vitro. For in vivo studies, rat models may have an advantage over mouse models for the study of neurodegeneration, as rat brain can demonstrate robust deposition of endogenously-expressed Aβ and tau in certain pathological states. Mouse models generally do not show the accumulation of insoluble, endogenously-expressed (non-transgenic) tau or Aβ. Therefore, to study neurodegeneration in CS and the possibility of Aβ and tau pathology, we generated an NHE6-null rat model of CS using CRISPR-Cas9 genome-editing. Here, we present the sequence of pathogenic events in neurodegenerating NHE6-null male rat brains across the lifespan. NHE6-null rats demonstrate an early and rapid loss of Purkinje cells in the cerebellum, as well as a more protracted neurodegenerative course in the cerebrum. In both the cerebellum and cerebrum, lysosome deficiency is an early pathogenic event, preceding autophagic dysfunction. Microglial and astrocyte activation also occur early. In the hippocampus and cortex, lysosome defects precede loss of pyramidal cells. Importantly, we subsequently observe biochemical and in situ evidence of both Aβ and tau aggregation in the aged NHE6-null hippocampus and cortex (but not in the cerebellum). Tau deposition is widely distributed, including cortical and subcortical distributions. Interestingly, we observe tau deposition in both neurons and glia, as has been reported in CS postmortem studies previously. In summary, this experimental model is among very few examples of a genetically modified animal that exhibits neurodegeneration with deposition of endogenously-expressed Aβ and tau. This NHE6-null rat will serve as a new robust model for CS. Furthermore, these studies provide evidence for linkages between endo-lysosome dysfunction and neurodegeneration involving protein aggregations, including Aβ and tau. Therefore these studies may provide insight into mechanisms of more common neurodegenerative disorders, including Alzheimer's Disease and related dementias.
    Keywords:  amyloid beta; lysosomes; neurodegeneration; rat model; tau
    DOI:  https://doi.org/10.1093/brain/awab467
  74. Cells. 2021 Dec 08. pii: 3464. [Epub ahead of print]10(12):
    The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma.
    Nicholas Nikesitch, Patricia Rebeiro, Lye Lin Ho, Srinivasa Pothula, Xin Maggie Wang, Tiffany Khong, Hazel Quek, Andrew Spencer, Cheok Soon Lee, Tara L Roberts, Silvia C W Ling.
      Background: Multiple myeloma (MM) remains incurable despite high-dose chemotherapy, autologous stem cell transplants and novel agents. Even with the improved survival of MM patients treated with novel agents, including bortezomib (Bz), the therapeutic options in relapsed/refractory MM remain limited. The majority of MM patients eventually develop resistance to Bz, although the mechanisms of the resistance are poorly understood. Methods: Lysosomal associated membrane protein 2A (LAMP2A) mRNA and protein expression levels were assessed in ex vivo patient samples and a Bz-resistant MM cell line model by in real-rime PCR, western blotting and immunohistochemistry. In vitro modelling of chaperone-mediated autophagy (CMA) activity in response to ER stress were assessed by western blotting and confocal microscopy. The effects of CMA inhibition on MM cell viability and Bz sensitivity in MM cells were assessed by Annexin V/7AAD apoptosis assays using flow cytometry. Results: In this study, there is evidence that CMA, a chaperone-mediated protein degradation pathway, is upregulated in Bz-resistant MM and the inhibition of CMA sensitises resistant cells to Bz. The protein levels of LAMP2A, the rate-limiting factor of the CMA pathway, are significantly increased in MM patients resistant to Bz and within our Bz-resistant cell line model. Bz-resistant cell lines also possessed higher basal CMA activity than the Bz-sensitive parent cell line. In MM cell lines, CMA activity was upregulated in response to ER stress induced by Bz. The inhibition of CMA sensitises Bz-resistant cells to Bz and the combination of CMA inhibition and Bz in vitro had a more cytotoxic effect on myeloma cells than Bz alone. Conclusion: In summary, the upregulation of CMA is a potential mechanism of resistance to Bz and a novel target to overcome Bz-resistant MM.
    Keywords:  ER stress and autophagy; LAMP2A; bortezomib; chaperone mediated autophagy; multiple myeloma; resistance
    DOI:  https://doi.org/10.3390/cells10123464
  75. Toxicology. 2021 Dec 21. pii: S0300-483X(21)00404-2. [Epub ahead of print] 153082
    The role of DRP1- PINK1-Parkin-mediated mitophagy in early cadmium-induced liver damage.
    Jian Sun, Fan Yu, Tao Wang, Jianchun Bian, Zongping Liu, Hui Zou.
      Cadmium (Cd) is an important environmental pollutant that causes varying degrees of damage to multiple systems of the body. However, the specific mechanism of Cd-induced liver mitophagy remains unclear. In the present study, 5-week-old BALB/c mice and a mouse liver parenchyma cell line (AML12) were studied using a combination of in vivo and in vitro studies. We found that Cd damaged liver cells, destroy the structure and function of mitochondria, and increased the production of superoxide anions. This study further examined the effect of Cd on mitochondrial dynamics and mitophagy and showed that Cd increased mitochondrial division and induced mitophagy. The PINK1-Parkin pathway is a classical mitophagy pathway. Cd-induced mitophagy was inhibited after significantly knocking down Pink1. Mdivi-1 can effectively inhibit mitochondrial division. In this study, Mdivi-1 inhibited the expression of DRP1 and significantly inhibited the occurrence of mitophagy induced by Cd. We further examined the effect of Cd on mitophagy flux. Cd did not increase lysosomal colocalization with mitochondria. In summary, Cd increase the level of oxidative stress, destroy the structure and function of mitochondria, destroy the homeostasis of mitochondrial division and fusion, induce mitophagy through the PINK1-Parkin pathway. Mitophagy plays a protective role in early cadmium-induced liver damage.
    Keywords:  Cadmium; DRP1; Liver; Mitochondria; Mitophagy; PINK1; Parkin
    DOI:  https://doi.org/10.1016/j.tox.2021.153082
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