bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2022–06–12
63 papers selected by
Viktor Korolchuk, Newcastle University
  1. The retroviral restriction factor TRIM5/TRIM5α regulates mitochondrial quality control.
  2. A unifying model for the role of the ATG8 system in autophagy.
  3. Emerging roles of mitotic autophagy.
  4. Autophagy at the intersection of aging, senescence, and cancer.
  5. The Genetics of Autophagy in Multicellular Organisms.
  6. TRIM27 cooperates with STK38L to inhibit ULK1-mediated autophagy and promote tumorigenesis.
  7. Contemporary mTOR inhibitor scaffolds to diseases breakdown: A patent review (2015-2021).
  8. The ORF7a protein of SARS-CoV-2 initiates autophagy and limits autophagosome-lysosome fusion via degradation of SNAP29 to promote virus replication.
  9. Mitochondrial Dysfunction in Spinocerebellar Ataxia Type 3 Is Linked to VDAC1 Deubiquitination.
  10. Mitochondria ROS and mitophagy in acute kidney injury.
  11. High glutamine suppresses osteogenesis through mTORC1-mediated inhibition of the mTORC2/AKT-473/RUNX2 axis.
  12. Autophagy is required for spermatogonial differentiation in the Drosophila testis.
  13. The Regulation of MiTF/TFE Transcription Factors Across Model Organisms: from Brain Physiology to Implication for Neurodegeneration.
  14. Inhibition of miR-421 Preserves Mitochondrial Function and Protects against Parkinson's Disease Pathogenesis via Pink1/Parkin-Dependent Mitophagy.
  15. Diacylglycerol kinase alleviates autophagic degradation of the endoplasmic reticulum in SPT10-deficient yeast to enhance triterpene biosynthesis.
  16. Lipoic acid blocks autophagic flux and impairs cellular bioenergetics in breast cancer and reduces stemness.
  17. The role of MiT/TFE family members in autophagy regulation.
  18. STAT3 suppresses the AMPKα/ULK1-dependent induction of autophagy in glioblastoma cells.
  19. Deletion of Rheb1 in Osteocytes Leads to Osteopenia Characterized by Reduced Bone Formation and Enhanced Bone Resorption.
  20. A Review of ApoE4 Interference Targeting Mitophagy Molecular Pathways for Alzheimer's Disease.
  21. Mitophagy in Alzheimer's disease: Molecular defects and therapeutic approaches.
  22. Feeding activates FGF15-SHP-TFEB-mediated lipophagy in the gut.
  23. Stratifin (SFN) regulates lung cancer progression via nucleating the Vps34-BECN1-TRAF6 complex for autophagy induction.
  24. Itaconate is a lysosomal inducer that promotes antibacterial innate immunity.
  25. Microglial mTOR Activation Upregulates Trem2 and Enhances β-Amyloid Plaque Clearance in the 5XFAD Alzheimer's Disease Model.
  26. Loss of 4E-BP converts cerebellar long-term depression to long-term potentiation.
  27. LC3-Mediated Mitophagy After CCCP or Vibrio splendidus Exposure in the Pacific Oyster Crassostrea gigas.
  28. Quercetin Can Improve Spinal Cord Injury by Regulating the mTOR Signaling Pathway.
  29. De Novo Development of Mitochondria-Targeted Molecular Probes Targeting Pink1.
  30. OPTN attenuates the neurotoxicity of abnormal Tau protein by restoring autophagy.
  31. Robustness of the Autophagy Pathway to Somatic Copy Number Losses.
  32. Tissue factor promotes HCC carcinogenesis by inhibiting BCL2-dependent autophagy.
  33. The Potential of Epigallocatechin Gallate (EGCG) in Targeting Autophagy for Cancer Treatment: A Narrative Review.
  34. Autophagic pathway contributes to low-nitrogen tolerance by optimizing nitrogen uptake and utilization in tomato.
  35. Ube2c-inhibition alleviated amyloid pathology and memory deficits in APP/PS1 mice model of AD.
  36. Lysosome lipid signalling from the periphery to neurons regulates longevity.
  37. Tonic inhibition of the chloride/proton antiporter ClC-7 by PI(3,5)P2 is crucial for lysosomal pH maintenance.
  38. A pivotal role of selective autophagy in mitochondrial quality control: Implications for zinc oxide nanoparticles induced neurotoxicity.
  39. Triphenyl phosphate (TPP) promotes hepatocyte toxicity via induction of endoplasmic reticulum stress and inhibition of autophagy flux.
  40. The Common Cellular Events in the Neurodegenerative Diseases and the Associated Role of Endoplasmic Reticulum Stress.
  41. Molecular basis of mEAK7-mediated human V-ATPase regulation.
  42. Major construction entails major demolition.
  43. Knockdown of miRNA-134-5p rescues dendritic deficits by promoting AMPK-mediated mitophagy in a mouse model of depression.
  44. Extracellular Vesicles as Therapy for CDH-associated Pulmonary Hypoplasia: Extra! Extra! Read All About Autophagy!
  45. Central role for p62/SQSTM1 in the elimination of toxic tau species in a mouse model of tauopathy.
  46. Lysosomal Proteomics Links Disturbances in Lipid Homeostasis and Sphingolipid Metabolism to CLN5 Disease.
  47. An Overview of Autophagy in Hematopoietic Stem Cell Transplantation.
  48. Effects of Individual Amino Acids on PPARα Transactivation, mTORC1 Activation, ApoA-I Transcription and pro-ApoA-I Secretion.
  49. Promising autophagy inhibitors: Therapeutic implications in oral cancer.
  50. Protective effect of the tunneling nanotube-TNFAIP2/M-sec system on podocyte autophagy in diabetic nephropathy.
  51. Emerging role of LETM1/GRP78 axis in lung cancer.
  52. From cradle to grave: neurogenesis, neuroregeneration and neurodegeneration in Alzheimer's and Parkinson's diseases.
  53. BAX regulates dendritic spine development via mitochondrial fusion.
  54. Evaluation of targeting autophagy for the treatment of malignant rhabdoid tumours.
  55. ERAS Is Constitutively Expressed in the Tissues of Adult Horses and May Be a Key Player in Basal Autophagy.
  56. An Update to Hallmarks of Cancer.
  57. Endolysosome Iron Chelation Inhibits HIV-1 Protein-Induced Endolysosome De-Acidification-Induced Increases in Mitochondrial Fragmentation, Mitophagy, and Cell Death.
  58. Preclinical and clinical evaluation of the LRRK2 inhibitor DNL201 for Parkinson's disease.
  59. Stress- and metabolic responses of Candida albicans require Tor1 kinase N-terminal HEAT repeats.
  60. The protective effect of selenoprotein M on non-alcoholic fatty liver disease: the role of the AMPKα1-MFN2 pathway and Parkin mitophagy.
  61. Clustering analysis revealed the autophagy classification and potential autophagy regulators' sensitivity of pancreatic cancer based on multi-omics data.
  62. Autophagy Impairment in App Knock-in Alzheimer's Model Mice.
  63. The Role of Autophagy and Pyroptosis in Liver Disorders.
  1. Autophagy. 2022 Jun 05. 1-2
    The retroviral restriction factor TRIM5/TRIM5α regulates mitochondrial quality control.
    Bhaskar Saha, Michael A Mandell.
      The protein TRIM5 is under intensive investigation related to its roles in antiviral defense, yet its underlying mechanisms of action remain elusive. In our study, we performed an unbiased identification of TRIM5-interacting partners and found proteins participating in a wide variety of cellular functions. We utilized this proteomics data set to uncover a role for TRIM5 in mitophagy, a mitochondrial quality control system that is impaired in multiple human diseases. Mitochondrial damage triggers the recruitment of TRIM5 to ER-mitochondria contact sites where TRIM5 colocalizes with markers of autophagosome biogenesis. Cells lacking TRIM5 are unable to carry out PRKN-dependent and PRKN-independent mitophagy pathways. TRIM5 knockout cells show reduced mitochondrial function and uncontrolled immune activation in response to mitochondrial damage; phenotypes consistent with a requirement for TRIM5 in mitophagy. Mechanistically, we found that TRIM5 is required for the recruitment of the autophagy initiation machinery to damaged mitochondria, where TRIM5 acts as a scaffold promoting interactions between protein markers of mitochondrial damage and the autophagy initiation machinery.
    Keywords:  APEX2; HIV-1; TRIM5α; autophagy; inflammation; mitochondria; mitophagy; restriction factor; tripartite-motif
    DOI:  https://doi.org/10.1080/15548627.2022.2084863
  2. J Cell Sci. 2022 Jun 01. pii: jcs258997. [Epub ahead of print]135(11):
    A unifying model for the role of the ATG8 system in autophagy.
    Thanh Ngoc Nguyen, Michael Lazarou.
      The formation of autophagosomes and their fusion with lysosomes are key events that underpin autophagic degradation of cargoes. The core ATG8 system, which consists of the ATG8 family of ubiquitin-like proteins and the machineries that conjugate them onto autophagosomal membranes, are among the most-studied autophagy components. Despite the research focus on the core ATG8 system, there are conflicting reports regarding its essential roles in autophagy. Here, we reconcile prior observations of the core ATG8 system into a unifying model of their function that aims to consider apparently conflicting discoveries. Bypass pathways of autophagy that function independently of the core ATG8 system are also discussed.
    Keywords:  ATG8; Autophagosome; Autophagy; GABARAP; LC3
    DOI:  https://doi.org/10.1242/jcs.258997
  3. J Cell Sci. 2022 Jun 01. pii: jcs255802. [Epub ahead of print]135(11):
    Emerging roles of mitotic autophagy.
    Eugenia Almacellas, Caroline Mauvezin.
      Lysosomes exert pleiotropic functions to maintain cellular homeostasis and degrade autophagy cargo. Despite the great advances that have boosted our understanding of autophagy and lysosomes in both physiology and pathology, their function in mitosis is still controversial. During mitosis, most organelles are reshaped or repurposed to allow the correct distribution of chromosomes. Mitotic entry is accompanied by a reduction in sites of autophagy initiation, supporting the idea of an inhibition of autophagy to protect the genetic material against harmful degradation. However, there is accumulating evidence revealing the requirement of selective autophagy and functional lysosomes for a faithful chromosome segregation. Degradation is the most-studied lysosomal activity, but recently described alternative functions that operate in mitosis highlight the lysosomes as guardians of mitotic progression. Because the involvement of autophagy in mitosis remains controversial, it is important to consider the specific contribution of signalling cascades, the functions of autophagic proteins and the multiple roles of lysosomes, as three entangled, but independent, factors controlling genomic stability. In this Review, we discuss the latest advances in this area and highlight the therapeutic potential of targeting autophagy for drug development.
    Keywords:  Autophagy; Cancer; Chromosomal instability; Chromosome segregation; Lysosomes; Mitosis
    DOI:  https://doi.org/10.1242/jcs.255802
  4. Mol Oncol. 2022 Jun 10.
    Autophagy at the intersection of aging, senescence, and cancer.
    Liam D Cassidy, Masashi Narita.
      Autophagy is an evolutionarily conserved cellular process in which macromolecules undergo lysosomal degradation. It fulfils essential roles in quality controlling cellular constituents and in energy homeostasis. Basal autophagy is also widely accepted to provide a protective role in aging and aging-related disorders, and its decline with age might precipitate the onset of a variety of diseases. In this review, we discuss the role of basal autophagy in maintaining homeostasis, in part through the maintenance of stem cell populations and the prevention of cellular senescence. We also consider how stress-induced senescence, for example during oncogene activation and in premalignant disease, might rely on autophagy, and the possibility that the age-associated decline in autophagy might promote tumor development through a variety of mechanisms. Ultimately, evidence suggests that autophagy is required for malignant cancer progression in a number of settings. Thus, autophagy appears to be tumor-suppressive during the early stages of tumorigenesis and tumor-promoting at later stages.
    Keywords:  Autophagy; aging; cancer; senescence
    DOI:  https://doi.org/10.1002/1878-0261.13269
  5. Annu Rev Genet. 2022 Jun 09.
    The Genetics of Autophagy in Multicellular Organisms.
    Hong Zhang.
      Autophagy, a lysosome-mediated degradation process evolutionarily conserved from yeast to mammals, is essential for maintaining cellular homeostasis and combating diverse cellular stresses. Autophagy involves de novo synthesis of a double-membrane autophagosome, sequestration of selected cellular contents, and subsequent delivery of sequestrated contents to the vacuole (in yeasts and plants) or to lysosomes (in animal cells) for degradation and recycling. Genetic studies in unicellular and multicellular model organisms have systematically revealed the molecular machinery, regulation, and function of autophagy in physiological settings. I review genetic studies in model organisms-from yeast to worm to fly-that enable us to not only identify autophagy genes, including ATG genes and the metazoan-specific EPG genes, but also uncover variants of autophagy in developmental contexts, novel regulatory mechanisms, and signaling events involved in mediating systemic autophagy response. Genetic analysis also helps us understand the liquid-liquid phase separation and transition that control autophagic degradation of protein aggregates. The emerging role of autophagy in zebrafish tissue regeneration is also discussed. Expected final online publication date for the Annual Review of Genetics, Volume 56 is November 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.
    DOI:  https://doi.org/10.1146/annurev-genet-022422-095608
  6. EMBO J. 2022 Jun 07. e109777
    TRIM27 cooperates with STK38L to inhibit ULK1-mediated autophagy and promote tumorigenesis.
    Yi Yang, Yifu Zhu, Shuai Zhou, Peipei Tang, Ran Xu, Yuwei Zhang, Dongping Wei, Jian Wen, Rick F Thorne, Xu Dong Zhang, Jun-Lin Guan, Lianxin Liu, Mian Wu, Song Chen.
      Autophagy represents a fundamental mechanism for maintaining cell survival and tissue homeostasis in response to physiological and pathological stress. Autophagy initiation converges on the FIP200-ATG13-ULK1 complex wherein the serine/threonine kinase ULK1 plays a central role. Here, we reveal that the E3 ubiquitin ligase TRIM27 functions as a negative regulatory component of the FIP200-ATG13-ULK1 complex. TRIM27 directly polyubiquitinates ULK1 at K568 and K571 sites with K48-linked ubiquitin chains, with proteasomal turnover maintaining control over basal ULK1 levels. However, during starvation-induced autophagy, TRIM27 catalyzes non-degradative K6- and K11-linked ubiquitination of the serine/threonine kinase 38-like (STK38L) kinase. In turn, STK38L ubiquitination promotes its activation and phosphorylation of ULK1 at Ser495, rendering ULK1 in a permissive state for TRIM27-mediated hyper-ubiquitination of ULK1. This cooperative mechanism serves to restrain the amplitude and duration of autophagy. Further evidence from mouse models shows that basal autophagy levels are increased in Trim27 knockout mice and that Trim27 differentially regulates tumorigenesis and metastasis. Our study identifies a key role of STK38L-TRIM27-ULK1 signaling axis in negatively controlling autophagy with relevance established in human breast cancer.
    Keywords:  STK38L; TRIM27; ULK1; autophagy; tumorigenesis
    DOI:  https://doi.org/10.15252/embj.2021109777
  7. Eur J Med Chem. 2022 May 28. pii: S0223-5234(22)00400-7. [Epub ahead of print]238 114498
    Contemporary mTOR inhibitor scaffolds to diseases breakdown: A patent review (2015-2021).
    Patrik Oleksak, Eugenie Nepovimova, Zofia Chrienova, Kamil Musilek, Jiri Patocka, Kamil Kuca.
      Mechanistic target of rapamycin (mTOR) is a highly conserved protein kinase acting as a central regulator of cell functions. The kinase forms two distinct mTOR complexes termed as mTORC1 and mTORC2. Dysregulation of mTOR activity is associated with various pathological conditions. Inhibition of overactivated mTOR represent a rational approach in the treatment of numerous human diseases. Rapamycin is a potent natural inhibitor of mTOR exhibiting significant antitumor and immunosuppressive activity. Derivatization of rapamycin provided rapalogs, the first generation of mTOR inhibitors that selectively inhibit mTORC1 activity. Further interest of research community resulted in creation of the second generation of mTOR inhibitors involving both, mTOR kinase inhibitors and dual phosphoinositide 3-kinase (PI3K)/mTOR inhibitors. Recently, combining advances of first and second generation of mTOR inhibitors yielded in the third generation of inhibitors termed as rapalinks. Nowadays, novel inhibitors belonging to all of the three generations are still under development. These inhibitors help us better to understand role of mTOR in mTOR signaling pathway as well as in diverse human diseases. In this review, we summarize recent reported mTOR inhibitors or methods of use thereof in the treatment of various diseases.
    Keywords:  Dual PI3K/mTOR inhibitors; PI3K/AKT/mTOR signaling pathway; Rapalinks; Rapalogs; Rapamycin; mTOR inhibitor; mTOR kinase inhibitors
    DOI:  https://doi.org/10.1016/j.ejmech.2022.114498
  8. Autophagy. 2022 Jun 07.
    The ORF7a protein of SARS-CoV-2 initiates autophagy and limits autophagosome-lysosome fusion via degradation of SNAP29 to promote virus replication.
    Peili Hou, Xuefeng Wang, Hongmei Wang, Tiecheng Wang, Zhangping Yu, Chunqing Xu, Yudong Zhao, Wenqi Wang, Yong Zhao, Fengyun Chu, Huasong Chang, Hongchao Zhu, Jiahui Lu, Fuzhen Zhang, Xue Liang, Xingyu Li, Song Wang, Yuwei Gao, Hongbin He.
      Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is closely related to various cellular aspects associated with autophagy. However, how SARS-CoV-2 mediates the subversion of the macroautophagy/autophagy pathway remains largely unclear. In this study, we demonstrate that overexpression of the SARS-CoV-2 ORF7a protein activates LC3-II and leads to the accumulation of autophagosomes in multiple cell lines, while knockdown of the viral ORF7a gene via shRNAs targeting ORF7a sgRNA during SARS-CoV-2 infection decreased autophagy levels. Mechanistically, the ORF7a protein initiates autophagy via the AKT-MTOR-ULK1-mediated pathway, but ORF7a limits the progression of autophagic flux by activating CASP3 (caspase 3) to cleave the SNAP29 protein at aspartic acid residue 30 (D30), ultimately impairing complete autophagy. Importantly, SARS-CoV-2 infection-induced accumulated autophagosomes promote progeny virus production, whereby ORF7a downregulates SNAP29, ultimately resulting in failure of autophagosome fusion with lysosomes to promote viral replication. Taken together, our study reveals a mechanism by which SARS-CoV-2 utilizes the autophagic machinery to facilitate its own propagation via ORF7a.
    Keywords:  Autophagosome-lysosome fusion; ORF7a; SARS-CoV-2; SNAP29; autophagy initiation; caspase activity
    DOI:  https://doi.org/10.1080/15548627.2022.2084686
  9. Int J Mol Sci. 2022 May 25. pii: 5933. [Epub ahead of print]23(11):
    Mitochondrial Dysfunction in Spinocerebellar Ataxia Type 3 Is Linked to VDAC1 Deubiquitination.
    Tina Harmuth, Jonasz J Weber, Anna J Zimmer, Anna S Sowa, Jana Schmidt, Julia C Fitzgerald, Ludger Schöls, Olaf Riess, Jeannette Hübener-Schmid.
      Dysfunctional mitochondria are linked to several neurodegenerative diseases. Metabolic defects, a symptom which can result from dysfunctional mitochondria, are also present in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease, the most frequent, dominantly inherited neurodegenerative ataxia worldwide. Mitochondrial dysfunction has been reported for several neurodegenerative disorders and ataxin-3 is known to deubiquitinylate parkin, a key protein required for canonical mitophagy. In this study, we analyzed mitochondrial function and mitophagy in a patient-derived SCA3 cell model. Human fibroblast lines isolated from SCA3 patients were immortalized and characterized. SCA3 patient fibroblasts revealed circular, ring-shaped mitochondria and featured reduced OXPHOS complexes, ATP production and cell viability. We show that wildtype ataxin-3 deubiquitinates VDAC1 (voltage-dependent anion channel 1), a member of the mitochondrial permeability transition pore and a parkin substrate. In SCA3 patients, VDAC1 deubiquitination and parkin recruitment to the depolarized mitochondria is inhibited. Increased p62-linked mitophagy, autophagosome formation and autophagy is observed under disease conditions, which is in line with mitochondrial fission. SCA3 fibroblast lines demonstrated a mitochondrial phenotype and dysregulation of parkin-VDAC1-mediated mitophagy, thereby promoting mitochondrial quality control via alternative pathways.
    Keywords:  Machado–Joseph disease; VDAC1 ubiquitination; ataxin-3; mitochondria dysfunction; spinocerebellar ataxia type 3
    DOI:  https://doi.org/10.3390/ijms23115933
  10. Autophagy. 2022 Jun 09. 1-14
    Mitochondria ROS and mitophagy in acute kidney injury.
    Lianjiu Su, Jiahao Zhang, Hernando Gomez, John A Kellum, Zhiyong Peng.
      Mitophagy is an essential mitochondrial quality control mechanism that eliminates damaged mitochondria and the production of reactive oxygen species (ROS). The relationship between mitochondria oxidative stress, ROS production and mitophagy are intimately interwoven, and these processes are all involved in various pathological conditions of acute kidney injury (AKI). The elimination of damaged mitochondria through mitophagy in mammals is a complicated process which involves several pathways. Furthermore, the interplay between mitophagy and different types of cell death, such as apoptosis, pyroptosis and ferroptosis in kidney injury is unclear. Here we will review recent advances in our understanding of the relationship between ROS and mitophagy, the different mitophagy pathways, the relationship between mitophagy and cell death, and the relevance of these processes in the pathogenesis of AKI.Abbreviations: AKI: acute kidney injury; AMBRA1: autophagy and beclin 1 regulator 1; ATP: adenosine triphosphate; BAK1: BCL2 antagonist/killer 1; BAX: BCL2 associated X, apoptosis regulator; BCL2: BCL2 apoptosis regulator; BECN1: beclin 1; BH3: BCL2 homology domain 3; BNIP3: BCL2 interacting protein 3; BNIP3L/NIX: BCL2 interacting protein 3 like; CASP1: caspase 1; CAT: catalase; CCCP: carbonyl cyanide m-chlorophenylhydrazone; CI-AKI: contrast-induced acute kidney injury; CISD1: CDGSH iron sulfur domain 1; CL: cardiolipin; CNP: 2',3'-cyclic nucleotide 3'-phosphodiesterase; DNM1L/DRP1: dynamin 1 like; E3: enzyme 3; ETC: electron transport chain; FA: folic acid; FUNDC1: FUN14 domain containing 1; G3P: glycerol-3-phosphate; G6PD: glucose-6-phosphate dehydrogenase; GPX: glutathione peroxidase; GSH: glutathione; GSK3B: glycogen synthase kinase 3 beta; GSR: glutathione-disulfide reductase; HIF1A: hypoxia inducible factor 1 subunit alpha; HUWE1: HECT, UBA and WWE domain containing 1; IL1B: interleukin 1 beta; IMM: inner mitochondrial membrane; IPC: ischemic preconditioning; IRI: ischemia-reperfusion injury; LIR: LC3-interacting region; LPS: lipopolysaccharide; MA: malate-aspartate; MPT: mitochondrial permeability transition; MUL1: mitochondrial E3 ubiquitin protein ligase 1; mtROS: mitochondrial ROS; NLR: NOD-like receptor; NLRP3: NLR family pyrin domain containing 3; NOX: NADPH oxidase; OGD-R: oxygen-glucose deprivation-reperfusion; OMM: outer mitochondrial membrane; OPA1: OPA1 mitochondrial dynamin like GTPase; OXPHOS: oxidative phosphorylation; PARL: presenilin associated rhomboid like; PINK1: PTEN induced kinase 1; PLSCR3: phospholipid scramblase 3; PMP: peptidase, mitochondrial processing; PRDX: peroxiredoxin; PRKN: parkin RBR E3 ubiquitin protein ligase; RPTC: rat proximal tubular cells; ROS: reactive oxygen species; SLC7A11/xCT: solute carrier family 7 member 11; SOD: superoxide dismutase; SOR: superoxide reductase; SQSTM1/p62: sequestosome 1; TCA: tricarboxylic acid; TIMM: translocase of inner mitochondrial membrane; TOMM: translocase of outer mitochondrial membrane; TXN: thioredoxin; VDAC: voltage dependent anion channel; VCP: valosin containing protein.
    Keywords:  Acute kidney injury; cell death; mitochondria; mitophagy; reactive oxygen species
    DOI:  https://doi.org/10.1080/15548627.2022.2084862
  11. Cell Death Discov. 2022 Jun 07. 8(1): 277
    High glutamine suppresses osteogenesis through mTORC1-mediated inhibition of the mTORC2/AKT-473/RUNX2 axis.
    Meher Bolisetti Gayatri, Navya Naidu Gajula, Suresh Chava, Aramati B M Reddy.
      Activation of the key nutrient cellular sensors mTORC1 and mTORC2 directs the fate of mesenchymal stromal cells (MSCs). Here, we report that glutamine regulates crosstalk between mTOR complexes and lineage commitment of MSCs independent of glucose concentration. High glutamine-induced mTORC1 hyperactivation resulted in the suppression of mTORC2, which otherwise stabilizes RUNX2 via GSK3β inhibition through pAKT-473. Activation of GSK3β resulted in the ubiquitination of RUNX2, a key transcription factor for the osteogenic commitment of MSCs. However, low glutamine conditions inhibit mTORC1 hyperactivation followed by increased mTORC2 activation and RUNX2 stabilization. Under diabetic/high-glucose conditions, glutamine-triggered hyperactivation of mTORC1 resulted in mTORC2 suppression, and active GSK3β led to suppression of RUNX2. Activation of p-AMPK by metformin inhibits high glutamine-induced mTORC1 hyperactivation and rescues RUNX2 through the mTORC2/AKT-473 axis. Collectively, our study indicates the role of glutamine in modulating MSC fate through cross-talk between mTOR complexes by identifying a critical switch in signaling. It also shows the importance of glutamine in modulating molecular cues (mTORC1/p-70S6K/mTORC2/RUNX2) that are involved in driving diabetes-induced bone adipogenesis and other secondary complications.
    DOI:  https://doi.org/10.1038/s41420-022-01077-3
  12. Biol Futur. 2022 Jun 07.
    Autophagy is required for spermatogonial differentiation in the Drosophila testis.
    Virginia B Varga, Dóra Schuller, Fanni Szikszai, Janka Szinyákovics, Gina Puska, Tibor Vellai, Tibor Kovács.
      Autophagy is a conserved, lysosome-dependent catabolic process of eukaryotic cells which is involved in cellular differentiation. Here, we studied its specific role in the differentiation of spermatogonial cells in the Drosophila testis. In the apical part of the Drosophila testis, there is a niche of germline stem cells (GSCs), which are connected to hub cells. Hub cells emit a ligand for bone morhphogenetic protein (BMP)-mediated signalling that represses Bam (bag of marbles) expression in GSCs to maintain them in an undifferentiated state. GSCs divide asymmetrically, and one of the daughter cells differentiates into a gonialblast, which eventually generates a cluster of spermatogonia (SG) by mitoses. Bam is active in SG, and defects in Bam function arrest these cells at mitosis. We show that BMP signalling represses autophagy in GSCs, but upregulates the process in SG. Inhibiting autophagy in SG results in an overproliferating phenotype similar to that caused by bam mutations. Furthermore, Bam deficiency leads to a failure in downstream mechanisms of the autophagic breakdown. These results suggest that the BMP-Bam signalling axis regulates developmental autophagy in the Drosophila testis, and that acidic breakdown of cellular materials is required for spermatogonial differentiation.
    Keywords:  Autophagy; BMP signalling pathway; Differentiation; Drosophila; Lysosome biogenesis; Stem cell
    DOI:  https://doi.org/10.1007/s42977-022-00122-7
  13. Mol Neurobiol. 2022 Jun 04.
    The Regulation of MiTF/TFE Transcription Factors Across Model Organisms: from Brain Physiology to Implication for Neurodegeneration.
    Francesco Agostini, Rossella Agostinis, Diego L Medina, Marco Bisaglia, Elisa Greggio, Nicoletta Plotegher.
      The microphthalmia/transcription factor E (MiTF/TFE) transcription factors are responsible for the regulation of various key processes for the maintenance of brain function, including autophagy-lysosomal pathway, lipid catabolism, and mitochondrial homeostasis. Among them, autophagy is one of the most relevant pathways in this frame; it is evolutionary conserved and crucial for cellular homeostasis. The dysregulation of MiTF/TFE proteins was shown to be involved in the development and progression of neurodegenerative diseases. Thus, the characterization of their function is key in the understanding of the etiology of these diseases, with the potential to develop novel therapeutics targeted to MiTF/TFE proteins and to the autophagic process. The fact that these proteins are evolutionary conserved suggests that their function and dysfunction can be investigated in model organisms with a simpler nervous system than the mammalian one. Building not only on studies in mammalian models but also in complementary model organisms, in this review we discuss (1) the mechanistic regulation of MiTF/TFE transcription factors; (2) their roles in different regions of the central nervous system, in different cell types, and their involvement in the development of neurodegenerative diseases, including lysosomal storage disorders; (3) the overlap and the compensation that occur among the different members of the family; (4) the importance of the evolutionary conservation of these protein and the process they regulate, which allows their study in different model organisms; and (5) their possible role as therapeutic targets in neurodegeneration.
    Keywords:  Autophagy; Lysosomal storage disorders; MiTF/TFE; Neurodegeneration; TFEB
    DOI:  https://doi.org/10.1007/s12035-022-02895-3
  14. Dis Markers. 2022 ;2022 5186252
    Inhibition of miR-421 Preserves Mitochondrial Function and Protects against Parkinson's Disease Pathogenesis via Pink1/Parkin-Dependent Mitophagy.
    Xiaolin Dong, Xianghua He, Li Yang, Qingyun Li, Yanming Xu.
      Mutations in PINK1 and Parkin are a major cause of Parkinson's disease (PD) pathogenesis. In addition, PINK1 and Parkin are two mitochondrial proteins that jointly contribute to mitochondrial homeostasis via mitophagy. Mitochondrial dysfunction is the most significant mechanism underlying PD pathogenesis. Thus, understanding the regulatory mechanism of PINK1 and Parkin expression is beneficial to the treatment of PD. In this study, we found that miR-421 expression was upregulated in mice treated with MPTP, as well as in SH-SY5Y cells treated with methyl-4-phenylpyridine (MPP+). Inhibition of miR-421 alleviated neurodegeneration in MPTP-treated mice and promoted mitophagy in MPP+-treated SH-SY5Y cells. Bioinformatics software predicted that Pink1 is a downstream target protein of miR-421. In addition, miR-421-induced Pink1 and Parkin inhibition negatively modulates mitophagy in MPP+-treated SH-SY5Y cells. In addition, our study confirmed that Pink1/Parkin is responsible for miR-421-regulated cell mitophagy. Overall, this study revealed that miR-421 regulates nerve cell mitophagy through the Pink1/Parkin pathway.
    DOI:  https://doi.org/10.1155/2022/5186252
  15. FEBS Lett. 2022 Jun 05.
    Diacylglycerol kinase alleviates autophagic degradation of the endoplasmic reticulum in SPT10-deficient yeast to enhance triterpene biosynthesis.
    Poornima Ramani Ranganathan, Ananth Krishna Narayanan, Niveditha Nawada, Monala Jayaprakash Rao, Kalyani Sai Reju, S Chaithra Priya, Tejal Gujarathi, Ravi Manjithaya, D K Venkata Rao.
      A recent study showed that deletion of the gene encoding the transcription regulator SuPpressor of Ty10 (SPT10) increases total phospholipids, and our previous study established a critical link between phospholipids and the mevalonate/ergosterol (MEV/ERG) pathway, which synthesizes triterpenes. This study aims to use spt10Δ yeast to improve triterpene production. Though MEV/ERG pathway was highly expressed in spt10Δ yeast, results showed insufficient accumulation of key metabolites and also revealed massive endoplasmic reticulum (ER) degradation. We found a stable, massive ER structure when we overexpressed diacylglycerol kinase1 (DGK1OE ) in spt10Δ yeast. Analyses of ER-stress and autophagy suggest that DGK1OE in the spt10Δ strain decreased autophagy, resulting in increased MEV/ERG pathway activity. Heterologous expression of β-amyrin synthase showed significant production of the triterpene β-amyrin in DGK1OE spt10Δ yeast. Overall, our study provides a strategic approach to improve triterpene production by increasing ER biogenesis while limiting ER degradation.
    Keywords:  Autophagy; ER stress; Endoplasmic reticulum; SPT10; Target of rapamycin (TOR) complex; diacylglycerol kinase1; synthetic biology
    DOI:  https://doi.org/10.1002/1873-3468.14418
  16. Biochim Biophys Acta Mol Basis Dis. 2022 Jun 06. pii: S0925-4439(22)00125-9. [Epub ahead of print] 166455
    Lipoic acid blocks autophagic flux and impairs cellular bioenergetics in breast cancer and reduces stemness.
    Bandana Chakravarti, Swati Rajput, Sana Raza, Sangam Rajak, Archana Tewari, Pratima Gupta, Aditya Upadhyay, Naibedya Chattopadhyay, Rohit Anthony Sinha.
      Autophagy inhibition is currently considered a novel therapeutic strategy for cancer treatment. Lipoic acid (LA), a naturally occurring compound found in all prokaryotic and eukaryotic cells, inhibits breast cancer cell growth; however, the effect of LA on autophagy-mediated breast cell death remains unknown. Our study identified that LA blocks autophagic flux by inhibiting autophagosome-lysosome fusion and lysosome activity which increases the accumulation of autophagosomes in MCF-7 and MDA-MB231 cells, leading to cell death of breast cancer cells. Interestingly, autophagic flux blockade limits the recycling of cellular fuels, resulting in insufficient substrates for cellular bioenergetics. Therefore, LA impairs cellular bioenergetics by the inhibition of mitochondrial function and glycolysis. We showed that LA-induced ROS generation is responsible for the blockade of autophagic flux and cellular bioenergetics in breast cancer cells. Moreover, LA-mediated blockade of autophagic flux and ROS generation may interfere with the regulation of the BCSCs/progenitor phenotype. Here, we demonstrate that LA inhibits mammosphere formation and subpopulation of BCSCs. Together, these results implicate that LA acts as a prooxidant, potent autophagic flux inhibitor, and causes energetic impairment which may lead to cell death of breast cancer cells/BCSCs.
    Keywords:  Autophagic flux; Autophagy; Breast Cancer stem cells; Breast cancer; Energy metabolism
    DOI:  https://doi.org/10.1016/j.bbadis.2022.166455
  17. Curr Top Biochem Res. 2021 ;22 151-159
    The role of MiT/TFE family members in autophagy regulation.
    Nicholas Theodosakis, Angel D Pagan, David E Fisher.
      The MiT/TFE family of proteins are important regulators of a number of metabolic processes. One of their most important roles is activating the autophagy pathway in the setting of nutrient deprivation or buildup of toxic metabolites. Their proper and improper functioning in this role has been linked to several types of disease, including cancer and multiple forms of neurodegeneration. In this review we will briefly outline what is known about individual family members' roles in regulating autophagy across a variety of contexts.
    Keywords:  Alzheimer’s; MITF; MiT/TFE; Parkinson’s; TFE3; TFEB; autophagy; neurodegeneration; pigmentation; tanning
  18. J Cell Mol Med. 2022 Jun 06.
    STAT3 suppresses the AMPKα/ULK1-dependent induction of autophagy in glioblastoma cells.
    Sujoy Bhattacharya, Jinggang Yin, Chuanhe Yang, Yinan Wang, Michelle Sims, Lawrence M Pfeffer, Edward Chaum.
      Despite advances in molecular characterization, glioblastoma (GBM) remains the most common and lethal brain tumour with high mortality rates in both paediatric and adult patients. The signal transducer and activator of transcription 3 (STAT3) is an important oncogenic driver of GBM. Although STAT3 reportedly plays a role in autophagy of some cells, its role in cancer cell autophagy remains unclear. In this study, we found Serine-727 and Tyrosine-705 phosphorylation of STAT3 was constitutive in GBM cell lines. Tyrosine phosphorylation of STAT3 in GBM cells suppresses autophagy, whereas knockout (KO) of STAT3 increases ULK1 gene expression, increases TSC2-AMPKα-ULK1 signalling, and increases lysosomal Cathepsin D processing, leading to the stimulation of autophagy. Rescue of STAT3-KO cells by the enforced expression of wild-type (WT) STAT3 reverses these pathways and inhibits autophagy. Conversely, expression of Y705F- and S727A-STAT3 phosphorylation deficient mutants in STAT3-KO cells did not suppress autophagy. Inhibition of ULK1 activity (by treatment with MRT68921) or its expression (by siRNA knockdown) in STAT3-KO cells inhibits autophagy and sensitizes cells to apoptosis. Taken together, our findings suggest that serine and tyrosine phosphorylation of STAT3 play critical roles in STAT3-dependent autophagy in GBM, and thus are potential targets to treat GBM.
    Keywords:  Atg14; Beclin1; Caspase-3; LC3-I/LC3-II; Prom1/CD133; MRT68921; RAD001 (Everolimus); Sequestome/p62; autophagy flux; mTORC1/2; apoptosis; cathepsin D; tuberin/TSC2
    DOI:  https://doi.org/10.1111/jcmm.17421
  19. DNA Cell Biol. 2022 Jun 10.
    Deletion of Rheb1 in Osteocytes Leads to Osteopenia Characterized by Reduced Bone Formation and Enhanced Bone Resorption.
    Jun Yang, Wuju Zhang, Eryong Lai, Wen Liu, Pinglin Lai, Zhipeng Zou, Weidong Wang, Xiaochun Bai.
      Ras homologue enriched in brain 1 (Rheb1), an upstream activator of the mechanistic target of rapamycin complex 1 (mTORC1), is known to modulate various cellular processes. However, its impact on bone metabolism in vivo remains unknown. The study aimed at understanding the role of Rheb1 on bone homeostasis. We measured the serum parameters and performed histomorphometry, quantitative real-time polymerase chain reaction, and Western blotting, along with the generation of mouse gene knockout (KO) model, and conducted a microcomputed tomography analysis and tartrate-resistant acid phosphatase staining, to delineate the impacts of Rheb1 on bone homeostasis. In the Rheb1 KO mice, the results showed that Rheb1 KO caused significant damage to the bone microarchitecture, indicating that mTORC1 activity was essential for the regulation of bone homeostasis. Specifically, suppressed mineralization activity in primary osteoblasts and a decreased osteoblast number were observed in the Rheb1 KO mice, demonstrating that loss of Rheb1 led to impaired osteoblastic differentiation. Furthermore, the higher apoptotic ratio in Rheb1-null osteocytes could promote Tnfsf11 expression and lead to an increase in osteoclasts, indicating increased bone resorption activity in the KO mice. The findings confirmed that Rheb1 deletion in osteoblasts/osteocytes led to osteopenia due to impaired bone formation and enhanced bone resorption.
    Keywords:  Rheb1; mature osteoblast; osteocyte; osteopenia
    DOI:  https://doi.org/10.1089/dna.2021.0874
  20. Front Aging Neurosci. 2022 ;14 881239
    A Review of ApoE4 Interference Targeting Mitophagy Molecular Pathways for Alzheimer's Disease.
    Huiyi Chen, Feng Chen, Ying Jiang, Lu Zhang, Guizhen Hu, Furong Sun, Miaoping Zhang, Yao Ji, Yanting Chen, Gang Che, Xu Zhou, Yu Zhang.
      Alzheimer's disease (AD) is one of the major worldwide causes of dementia that is characterized by irreversible decline in learning, memory loss, and behavioral impairments. Mitophagy is selective autophagy through the clearance of aberrant mitochondria, specifically for degradation to maintain energy generation and neuronal and synaptic function in the brain. Accumulating evidence shows that defective mitophagy is believed to be as one of the early and prominent features in AD pathogenesis and has drawn attention in the recent few years. APOE ε4 allele is the greatest genetic determinant for AD and is widely reported to mediate detrimental effects on mitochondria function and mitophagic process. Given the continuity of the physiological process, this review takes the mitochondrial dynamic and mitophagic core events into consideration, which highlights the current knowledge about the molecular alterations from an APOE-genotype perspective, synthesizes ApoE4-associated regulations, and the cross-talk between these signaling, along with the focuses on general autophagic process and several pivotal processes of mitophagy, including mitochondrial dynamic (DRP1, MFN-1), mitophagic induction (PINK1, Parkin). These may shed new light on the link between ApoE4 and AD and provide novel insights for promising mitophagy-targeted therapeutic strategies for AD.
    Keywords:  Alzheimer's disease; apolipoprotein E; mitochondrial dynamics; mitophagy; neurodegenerative disease
    DOI:  https://doi.org/10.3389/fnagi.2022.881239
  21. Mol Psychiatry. 2022 Jun 03.
    Mitophagy in Alzheimer's disease: Molecular defects and therapeutic approaches.
    Arnaud Mary, Fanny Eysert, Frédéric Checler, Mounia Chami.
      Mitochondrial dysfunctions are central players in Alzheimer's disease (AD). In addition, impairments in mitophagy, the process of selective mitochondrial degradation by autophagy leading to a gradual accumulation of defective mitochondria, have also been reported to occur in AD. We provide an updated overview of the recent discoveries and advancements on mitophagic molecular dysfunctions in AD-derived fluids and cells as well as in AD brains. We discuss studies using AD cellular and animal models that have unraveled the contribution of relevant AD-related proteins (Tau, Aβ, APP-derived fragments and APOE) in mitophagy failure. In accordance with the important role of impaired mitophagy in AD, we report on various therapeutic strategies aiming at stimulating mitophagy in AD and we summarize the benefits of these potential therapeutic strategies in human clinical trials.
    DOI:  https://doi.org/10.1038/s41380-022-01631-6
  22. EMBO J. 2022 Jun 10. e109997
    Feeding activates FGF15-SHP-TFEB-mediated lipophagy in the gut.
    Sunmi Seok, Young-Chae Kim, Yang Zhang, Bo Kong, Grace Guo, Jian Ma, Byron Kemper, Jongsook Kim Kemper.
      Lysosome-mediated macroautophagy, including lipophagy, is activated under nutrient deprivation but is repressed after feeding. We show that, unexpectedly, feeding activates intestinal autophagy/lipophagy in a manner dependent on both the orphan nuclear receptor, small heterodimer partner (SHP/NR0B2), and the gut hormone, fibroblast growth factor-15/19 (FGF15/19). Furthermore, postprandial intestinal triglycerides (TGs) and apolipoprotein-B48 (ApoB48), the TG-rich chylomicron marker, were elevated in SHP-knockout and FGF15-knockout mice. Genomic analyses of the mouse intestine indicated that SHP partners with the key lysosomal activator, transcription factor-EB (TFEB) to upregulate the transcription of autophagy/lipolysis network genes after feeding. FGF19 treatment activated lipophagy, reducing TG and ApoB48 levels in HT29 intestinal cells, which was dependent on TFEB. Mechanistically, feeding-induced FGF15/19 signaling increased the nuclear localization of TFEB and SHP via PKC beta/zeta-mediated phosphorylation, leading to increased transcription of the TFEB/SHP target lipophagy genes, Ulk1 and Atgl. Collectively, these results demonstrate that paradoxically after feeding, FGF15/19-activated SHP and TFEB activate gut lipophagy, limiting postprandial TGs. As excess postprandial lipids cause dyslipidemia and obesity, the FGF15/19-SHP-TFEB axis that reduces intestinal TGs via lipophagic activation provides promising therapeutic targets for obesity-associated metabolic disease.
    Keywords:  ATGL; FGF19; SHP; TFEB; autophagy
    DOI:  https://doi.org/10.15252/embj.2021109997
  23. Clin Transl Med. 2022 Jun;12(6): e896
    Stratifin (SFN) regulates lung cancer progression via nucleating the Vps34-BECN1-TRAF6 complex for autophagy induction.
    Ji Young Kim, Mi-Jeong Kim, Ji Su Lee, Juhee Son, Duk-Hwan Kim, Joo Sang Lee, Soo-Kyung Jeong, Eunyoung Chun, Ki-Young Lee.
      
    DOI:  https://doi.org/10.1002/ctm2.896
  24. Mol Cell. 2022 May 25. pii: S1097-2765(22)00443-9. [Epub ahead of print]
    Itaconate is a lysosomal inducer that promotes antibacterial innate immunity.
    Zhenxing Zhang, Chao Chen, Fan Yang, Yi-Xin Zeng, Pengkai Sun, Ping Liu, Xinjian Li.
      Lysosomes are the main organelles in macrophages for killing invading bacteria. However, the precise mechanism underlying lysosomal biogenesis upon bacterial infection remains enigmatic. We demonstrate here that LPS stimulation increases IRG1-dependent itaconate production, which promotes lysosomal biogenesis by activating the transcription factor, TFEB. Mechanistically, itaconate directly alkylates human TFEB at cysteine 212 (Cys270 in mice) to induce its nuclear localization by antagonizing mTOR-mediated phosphorylation and cytosolic retention. Functionally, abrogation of itaconate synthesis by IRG1/Irg1 knockout or expression of an alkylation-deficient TFEB mutant impairs the antibacterial ability of macrophages in vitro. Furthermore, knockin mice harboring an alkylation-deficient TFEB mutant display elevated susceptibility to Salmonella typhimurium infection, whereas in vivo treatment of OI, a cell-permeable itaconate derivative, limits inflammation. Our study identifies itaconate as an endogenous metabolite that functions as a lysosomal inducer in macrophages in response to bacterial infection, implying the potential therapeutic utility of itaconate in treating human bacterial infection.
    Keywords:  TFEB; alkylation; bacterial infection; innate immunity; itaconate; lysosomal biogenesis; macrophage
    DOI:  https://doi.org/10.1016/j.molcel.2022.05.009
  25. J Neurosci. 2022 Jun 02. pii: JN-RM-2427-21. [Epub ahead of print]
    Microglial mTOR Activation Upregulates Trem2 and Enhances β-Amyloid Plaque Clearance in the 5XFAD Alzheimer's Disease Model.
    Qian Shi, Cheng Chang, Afaf Saliba, Manzoor A Bhat.
      The mechanistic Target of Rapamycin (mTOR) signaling pathway plays a major role in key cellular processes including metabolism and differentiation; however, the role of mTOR in microglia and its importance in Alzheimer's disease (AD) has remained largely uncharacterized. We report that selective loss of Tsc1, a negative regulator of mTOR, in microglia in mice of both sexes, caused mTOR activation and upregulation of Trem2 with enhanced β-Amyloid clearance, reduced spine loss, and improved cognitive function in the 5XFAD AD mouse model. Combined loss of Tsc1 and Trem2 in microglia led to reduced β-Amyloid clearance and increased Aβ plaque burden revealing that Trem2 functions downstream of mTOR. Tsc1 mutant microglia showed increased phagocytosis with upregulation of CD68 and Lamp1 lysosomal proteins. In vitro studies using Tsc1-deficient microglia revealed enhanced endocytosis of the lysosomal tracker indicator Green DND-26 suggesting increased lysosomal activity. Incubation of Tsc1-deficient microglia with fluorescent-labeled Aβ revealed enhanced Aβ uptake and clearance, which was blunted by rapamycin, an mTOR inhibitor. In vivo treatment of mice of relevant genotypes in the 5XFAD background with rapamycin, affected microglial activity, decreased Trem2 expression and reduced Aβ clearance causing an increase in Aβ plaque burden. Prolonged treatment with rapamycin caused even further reduction of mTOR activity, reduction in Trem2 expression, and increase in Aβ levels. Together, our findings reveal that mTOR signaling in microglia is critically linked to Trem2 regulation and lysosomal biogenesis, and that the up-regulation of Trem2 in microglia through mTOR activation could be exploited towards better therapeutic avenues to β-Amyloid-related AD pathologies.Significance statement:mTOR signaling pathway is a key regulator for major cellular metabolic processes. However, the link between mTOR signaling and AD is not well understood. In this study, we provide compelling in vivo evidence that mTOR activation in microglia would benefit β-Amyloid related AD pathologies, as it upregulates Trem2, a key receptor for β-Amyloid plaque uptake. Inhibition of mTOR pathway with rapamycin, a well-established immunosuppressant, downregulated Trem2 in microglia and reduced β-Amyloid plaque clearance indicating that mTOR inactivation may be detrimental in β-Amyloid-associated AD patients. This finding will have a significant public health impact and benefit, regarding the usage of rapamycin in AD patients, which we believe will aggravate the β-Amyloid-related AD pathologies.
    DOI:  https://doi.org/10.1523/JNEUROSCI.2427-21.2022
  26. Cell Rep. 2022 Jun 07. pii: S2211-1247(22)00688-X. [Epub ahead of print]39(10): 110911
    Loss of 4E-BP converts cerebellar long-term depression to long-term potentiation.
    Natasha Saviuk, Yumaine Chong, Peng Wang, Sara Bermudez, Zhe Zhao, Arjun A Bhaskaran, Derek Bowie, Nahum Sonenberg, Ellis Cooper, A Pejmun Haghighi.
      Genetic perturbances in translational regulation result in defects in cerebellar motor learning; however, little is known about the role of translational mechanisms in the regulation of cerebellar plasticity. We show that genetic removal of 4E-BP, a translational suppressor and target of mammalian target of rapamycin complex 1, results in a striking change in cerebellar synaptic plasticity. We find that cerebellar long-term depression (LTD) at parallel fiber-Purkinje cell synapses is converted to long-term potentiation in 4E-BP knockout mice. Biochemical and pharmacological experiments suggest that increased phosphatase activity largely accounts for the defects in LTD. Our results point to a model in which translational regulation through the action of 4E-BP plays a critical role in establishing the appropriate kinase/phosphatase balance required for normal synaptic plasticity in the cerebellum.
    Keywords:  4E-BP; CP: Neuroscience; LTD; PP2A; Purkinje cells; autism; cerebellum; mTOR; phosphatase kinase balance; synaptic plasticity; translation
    DOI:  https://doi.org/10.1016/j.celrep.2022.110911
  27. Front Cell Dev Biol. 2022 ;10 885478
    LC3-Mediated Mitophagy After CCCP or Vibrio splendidus Exposure in the Pacific Oyster Crassostrea gigas.
    Jiejie Sun, Xiaoqian Lv, Jinyuan Leng, Lingling Wang, Linsheng Song.
      Mitochondrial selective autophagy, known as mitophagy, surveils the mitochondrial population by eliminating superfluous and/or impaired organelles to mediate cellular survival and viability in response to injury/trauma and infection. In this study, the components of the mitophagy pathway in the Pacific oyster Crassostrea gigas were screened from NCBI with reference to the protein sequences of the human mitophagy process. A total of 10 mitophagy process-related genes were identified from C. gigas, including NIX, FUNDC1, PHB2, Cardiolipin, P62, VDAC2, MFN2, PARL, MPP, and OPTN. They shared high similarities with their homologs in the human mitophagy pathway and were expressed in various tissues of C. gigas. After CCCP exposure, the fluorescence intensity of the mitochondrial probe JC-1 monomers increased significantly in hemocytes, while the fluorescence intensity of JC-1 aggregates decreased significantly. Meanwhile, the fluorescence of lysosomes was found to be co-localized with that of CgLC3 and mitochondria in CCCP-treated hemocytes. Double- and single-membrane-bound vacuoles resembling autophagic structures were observed in the hemocytes after CCCP exposure. The fluorescence intensity of JC-1 monomers and the abundance of CgLC3Ⅱ in hemocytes both increased after Vibrio splendidus exposure. At the same time, the green signals of CgLC3 were co-localized with red signals of the mitochondria, and the fluorescence intensity of autophagy increased significantly in hemocytes after V. splendidus exposure. The results confirmed the existence of a complete mitophagy pathway in mollusks for the first time, which was helpful for further study on the function of mitochondrial autophagy in mollusks.
    Keywords:  CCCP; Crassostrea gigas; LC3; Vibrio splendidus; mitophagy
    DOI:  https://doi.org/10.3389/fcell.2022.885478
  28. Front Neurol. 2022 ;13 905640
    Quercetin Can Improve Spinal Cord Injury by Regulating the mTOR Signaling Pathway.
    Xichen Wang, Yuke Fu, Benson O A Botchway, Yufeng Zhang, Yong Zhang, Tian Jin, Xuehong Liu.
      The pathogenesis of spinal cord injury (SCI) is complex. At present, there is no effective treatment for SCI, with most current interventions focused on improving the symptoms. Inflammation, apoptosis, autophagy, and oxidative stress caused by secondary SCI may instigate serious consequences in the event of SCI. The mammalian target of rapamycin (mTOR), as a key signaling molecule, participates in the regulation of inflammation, apoptosis, and autophagy in several processes associated with SCI. Quercetin can reduce the loss of myelin sheath, enhance the ability of antioxidant stress, and promote axonal regeneration. Moreover, quercetin is also a significant player in regulating the mTOR signaling pathway that improves pathological alterations following neuronal injury. Herein, we review the therapeutic effects of quercetin in SCI through its modulation of the mTOR signaling pathway and elaborate on how it can be a potential interventional agent for SCI.
    Keywords:  apoptosis; autophagy; autophagy mTOR signaling pathway; inflammation; mTOR signaling pathway; quercetin; spinal cord injury
    DOI:  https://doi.org/10.3389/fneur.2022.905640
  29. Int J Mol Sci. 2022 May 28. pii: 6076. [Epub ahead of print]23(11):
    De Novo Development of Mitochondria-Targeted Molecular Probes Targeting Pink1.
    Shulamit Fluss Ben-Uliel, Faten Habrat Zoabi, Moriya Slavin, Hadas Sibony-Benyamini, Nir Kalisman, Nir Qvit.
      Mitochondria play central roles in maintaining cellular metabolic homeostasis, cell survival and cell death, and generate most of the cell's energy. Mitochondria maintain their homeostasis by dynamic (fission and fusion) and quality control mechanisms, including mitophagy, the removal of damaged mitochondria that is mediated mainly by the Pink1/Parkin pathway. Pink1 is a serine/threonine kinase which regulates mitochondrial function, hitherto many molecular mechanisms underlying Pink1 activity in mitochondrial homeostasis and cell fate remain unknown. Peptides are vital biological mediators that demonstrate remarkable potency, selectivity, and low toxicity, yet they have two major limitations, low oral bioavailability and poor stability. Herein, we rationally designed a linear peptide that targets Pink1 and, using straightforward chemistry, we developed molecular probes with drug-like properties to further characterize Pink1. Initially, we conjugated a cell-penetrating peptide and a cross-linker to map Pink1's 3D structure and its interaction sites. Next, we conjugated a fluorescent dye for cell-imaging. Finally, we developed cyclic peptides with improved stability and binding affinity. Overall, we present a facile approach to converting a non-permeable linear peptide into a research tool possessing important properties for therapeutics. This is a general approach using straightforward chemistry that can be tailored for various applications by numerous laboratories.
    Keywords:  Pink1; backbone cyclization; bioactive peptides; mitochondria; mitophagy; molecular probes; peptidomimetics; protein-peptide interactions; protein-protein interactions; therapeutic peptides
    DOI:  https://doi.org/10.3390/ijms23116076
  30. Transl Psychiatry. 2022 Jun 04. 12(1): 230
    OPTN attenuates the neurotoxicity of abnormal Tau protein by restoring autophagy.
    Yin Xu, Yun Liu, Xi Chen, Qia Xu, Liwei Liu, Hui Liu, Ruowen Guo, Yide Qin.
      OPTN is an autophagy receptor involved in autophagic degradation. Here we studied the role of OPTN in attenuating the neurotoxicity induced by mutated Tau protein. We constructed recombinant adeno-associated viruses with OPTN and Tau-P301L genes, respectively. Through virus coinfection on neuronal cell line HT22 in vitro and Kunming mice in vivo, we found that autophagy- and apoptosis-associated genes are altered by Tau-P301L at both mRNA and protein levels, which are restored by OPTN expression. Functionally, OPTN suppresses apoptosis and enhances cellular viability in Tau-P301L expressing HT22 cells, and increases learning and memory in Tau-P301L expressing mice, respectively. Last, we found that OPTN reduces the p-Tau levels in vitro and in vivo. Our results reveal the function of OPTN in lowering the p-Tau level and the expressions of apoptosis genes, and increasing the expressions of autophagic genes, indicating a beneficial role of OPTN in Tau pathology.
    DOI:  https://doi.org/10.1038/s41398-022-02004-x
  31. Cells. 2022 May 27. pii: 1762. [Epub ahead of print]11(11):
    Robustness of the Autophagy Pathway to Somatic Copy Number Losses.
    Pierfrancesco Polo, Niklas Gremke, Thorsten Stiewe, Michael Wanzel.
      Autophagy allows cells to temporarily tolerate energy stress by replenishing critical metabolites through self-digestion, thereby attenuating the cytotoxic effects of anticancer drugs that target tumor metabolism. Autophagy defects could therefore mark a metabolically vulnerable cancer state and open a therapeutic window. While mutations of autophagy genes (ATGs) are notably rare in cancer, haploinsufficiency network analyses across many cancers have shown that the autophagy pathway is frequently hit by somatic copy number losses of ATGs such as MAP1LC3B/ATG8F (LC3), BECN1/ATG6 (Beclin-1), and ATG10. Here, we used CRISPR/Cas9 technology to delete increasing numbers of copies of one or more of these ATGs in non-small cell lung cancer cells and examined the effects on sensitivity to compounds targeting aerobic glycolysis, a hallmark of cancer metabolism. Whereas the complete knockout of one ATG blocked autophagy and led to profound metabolic vulnerability, this was not the case for combinations of different nonhomozygous deletions. In cancer patients, the effect of ATG copy number loss was blunted at the protein level and did not lead to the accumulation of p62 as a sign of reduced autophagic flux. Thus, the autophagy pathway is shown to be markedly robust and resilient, even with the concomitant copy number loss of key autophagy genes.
    Keywords:  aerobic glycolysis; autophagy; cancer metabolism; copy number loss; haploinsufficiency; metabolic cancer therapy; somatic copy number alterations (SCNAs)
    DOI:  https://doi.org/10.3390/cells11111762
  32. Bull Cancer. 2022 Jun 06. pii: S0007-4551(22)00164-3. [Epub ahead of print]
    Tissue factor promotes HCC carcinogenesis by inhibiting BCL2-dependent autophagy.
    Jia Liu, Bang Liu, Guanghao Diao, Zhiqiang Zhang.
       INTRODUCTION: Tissue factor (TF) is an important predictor for poor prognosis of Hepatocellular carcinoma (HCC). TF can also upregulate the expression of BCL2, which is a key inhibitor of autophagy responses. This study aims to explore the role of BCL2-dependent autophagy in TF-regulated HCC carcinogenesis.
    METHODS: In this study, we explored the roles of TF in HCC using gene overexpression and silencing assays. Besides, we further identified the significance of BCL2-Beclin1-autophagy signaling on TF-regulated HCC tumorigenesis by combining TF silencing with pharmacological autophagy inhibitor (3-MA).
    RESULTS: The experimental data showed that the overexpressed TF promoted BCL2 protein expression and inhibited the autophagy activity (shown as LC3 conversion rate, p62 expression and autophagosomes) while maintaining the survival in HCC cells. In contrast, the silent TF showed the completely opposite results. Furthermore, TF knockdown promoted the dissociation of Beclin1 from BCL2-Beclin1 complex. In addition, the enhanced autophagy and inhibited survival by TF knockdown could be reversed by autophagy inhibition with 3-MA or spautin-1 (Beclin1 specific inhibitor) in HCC cells. Xenografts assays also showed that TF-silencing HCC cells had stronger tumorigenicity in vivo, which was recovered by spautin-1 administration.
    CONCLUSIONS: TF inhibits autophagy-related death by enhancing BCL2 expression, whereby promoting HCC tumorigenesis.
    Keywords:  Tissue factor, Hepatocellular carcinoma, BCL2, Autophagy, Beclin1
    DOI:  https://doi.org/10.1016/j.bulcan.2022.04.007
  33. Int J Mol Sci. 2022 May 28. pii: 6075. [Epub ahead of print]23(11):
    The Potential of Epigallocatechin Gallate (EGCG) in Targeting Autophagy for Cancer Treatment: A Narrative Review.
    Elena Ferrari, Saverio Bettuzzi, Valeria Naponelli.
      Autophagy is an evolutionarily conserved process for the degradation of redundant or damaged cellular material by means of a lysosome-dependent mechanism, contributing to cell homeostasis and survival. Autophagy plays a multifaceted and context-dependent role in cancer initiation, maintenance, and progression; it has a tumor suppressive role in the absence of disease and is upregulated in cancer cells to meet their elevated metabolic demands. Autophagy represents a promising but challenging target in cancer treatment. Green tea is a widely used beverage with healthy effects on several diseases, including cancer. The bioactive compounds of green tea are mainly catechins, and epigallocatechin-gallate (EGCG) is the most abundant and biologically active among them. In this review, evidence of autophagy modulation and anti-cancer effects induced by EGCG treatment in experimental cancer models is presented. Reviewed articles reveal that EGCG promotes cytotoxic autophagy often through the inactivation of PI3K/Akt/mTOR pathway, resulting in apoptosis induction. EGCG pro-oxidant activity has been postulated to be responsible for its anti-cancer effects. In combination therapy with a chemotherapy drug, EGCG inhibits cell growth and the drug-induced pro-survival autophagy. The selected studies rightly claim EGCG as a valuable agent in cancer chemoprevention.
    Keywords:  autophagy; autophagy activator; autophagy modulator; cancer therapy; epigallocatechin gallate
    DOI:  https://doi.org/10.3390/ijms23116075
  34. Hortic Res. 2022 ;9 uhac068
    Autophagic pathway contributes to low-nitrogen tolerance by optimizing nitrogen uptake and utilization in tomato.
    Jiajian Cao, Xuelian Zheng, Dongling Xie, Hui Zhou, Shujun Shao, Jie Zhou.
      Autophagy is a primary process involved in the degradation and reuse of redundant or damaged cytoplasmic components in eukaryotes. Autophagy has been demonstrated to facilitate nutrient recycling and remobilization by delivering intracellular materials to the vacuole for degradation in plants under nutrient starvation. However, the role of autophagy in nitrogen (N) uptake and utilization remains unknown. Here, we report that the ATG6-dependent autophagic pathway regulates N utilization in tomato (Solanum lycopersicum) under low-nitrogen (LN) conditions. Autophagy-disrupted mutants exhibited weakened biomass production and N accumulation compared with wild-type (WT), while ATG6 overexpression promoted autophagy and biomass production under LN stress. The N content in atg6 mutants decreased while that in ATG6-overexpressing lines increased due to the control of N transporter gene expression in roots under LN conditions. Furthermore, ATG6-dependent autophagy enhanced N assimilation efficiency and protein production in leaves. Nitrate reductase and nitrite reductase activities and expression were compromised in atg6 mutants but were enhanced in ATG6-overexpressing plants under LN stress. Moreover, ATG6-dependent autophagy increased plant carbon fixation and photosynthetic capacity. The quantum yield of photosystem II, photosynthetic N use efficiency and photosynthetic protein accumulation were compromised in atg6 mutants but were restored in ATG6-overexpressing plants. A WT scion grafted onto atg6 mutant rootstock and an atg6 scion grafted onto WT rootstock both exhibited inhibited LN-induced autophagy and N uptake and utilization. Thus, ATG6-dependent autophagy regulates not only N uptake and utilization as well as carbon assimilation but also nutrient recycling and remobilization in tomato plants experiencing LN stress.
    DOI:  https://doi.org/10.1093/hr/uhac068
  35. Prog Neurobiol. 2022 Jun 04. pii: S0301-0082(22)00084-3. [Epub ahead of print] 102298
    Ube2c-inhibition alleviated amyloid pathology and memory deficits in APP/PS1 mice model of AD.
    Tian Li, Qiang Su, Zhi-Na Zhang, Yan-Li Zhang, Mingxuan Yang, Zhuoran Wang, Junhong Guo, Zhaojun Wang, Meina Wu, Hongyan Cai, Jinshun Qi.
      Autophagy is a major intracellular degradation pathway for the clearance of damaged organelles and misfolded peptides. Previous studies have indicated that autophagy is involved in the pathogenesis of neurodegenerative disease including Alzheimer's disease (AD). Defective autophagy and highly expressed ubiquitin-conjugating enzyme 2C (Ube2c) have been found in AD patients and mouse. However, little is known about the regulation of autophagy in AD. The association of Ube2c with autophagy, amyloid pathology and cognitive deficits in AD remains unclear. In the present study, we characterized over expression of Ube2c and declined autophagy in amyloid β (Aβ)-treated microglia and demonstrated the protective effects of agomelatine (AGO) in APP/PS1 mice. We found that knockdown of Ube2c with AAV2 encoding shUbe2c resulted in an obvious enhancement of autophagy in BV2 microglia cells, and an alleviation of Aβ pathology and memory deficits in APP/PS1 mice. Further, pharmacological inhibition of Ube2c by AGO significantly reduced Aβ plaques, improved synaptic plasticity and cognitive behaviors in APP/PS1 mice, as well as promoted autophagy in microglia. Our findings uncover a potent role of Ube2c over-expression and autophagy decline in the pathogenesis of AD, and suggest that regulation of Ube2c and autophagy may provide an important clue and a potential target for the novel therapeutics of AD.
    Keywords:  APP/PS1 mice; Agomelatine; Ube2c; autophagy; microglia
    DOI:  https://doi.org/10.1016/j.pneurobio.2022.102298
  36. Nat Cell Biol. 2022 Jun 09.
    Lysosome lipid signalling from the periphery to neurons regulates longevity.
    Marzia Savini, Andrew Folick, Yi-Tang Lee, Feng Jin, André Cuevas, Matthew C Tillman, Jonathon D Duffy, Qian Zhao, Isaiah A Neve, Pei-Wen Hu, Yong Yu, Qinghao Zhang, Youqiong Ye, William B Mair, Jin Wang, Leng Han, Eric A Ortlund, Meng C Wang.
      Lysosomes are key cellular organelles that metabolize extra- and intracellular substrates. Alterations in lysosomal metabolism are implicated in ageing-associated metabolic and neurodegenerative diseases. However, how lysosomal metabolism actively coordinates the metabolic and nervous systems to regulate ageing remains unclear. Here we report a fat-to-neuron lipid signalling pathway induced by lysosomal metabolism and its longevity-promoting role in Caenorhabditis elegans. We discovered that induced lysosomal lipolysis in peripheral fat storage tissue upregulates the neuropeptide signalling pathway in the nervous system to promote longevity. This cell-non-autonomous regulation is mediated by a specific polyunsaturated fatty acid, dihomo-γ-linolenic acid, and LBP-3 lipid chaperone protein transported from the fat storage tissue to neurons. LBP-3 binds to dihomo-γ-linolenic acid, and acts through NHR-49 nuclear receptor and NLP-11 neuropeptide in neurons to extend lifespan. These results reveal lysosomes as a signalling hub to coordinate metabolism and ageing, and lysosomal signalling mediated inter-tissue communication in promoting longevity.
    DOI:  https://doi.org/10.1038/s41556-022-00926-8
  37. Elife. 2022 Jun 07. pii: e74136. [Epub ahead of print]11
    Tonic inhibition of the chloride/proton antiporter ClC-7 by PI(3,5)P2 is crucial for lysosomal pH maintenance.
    Xavier Leray, Jacob K Hilton, Kamsi Nwangwu, Alissa Becerril, Vedrana Mikusevic, Gabriel Fitzgerald, Anowarul Amin, Mary R Weston, Joseph A Mindell.
      The acidic luminal pH of lysosomes, maintained within a narrow range, is essential for proper degrative function of the organelle and is generated by the action of a V-type H+ ATPase, but other pathways for ion movement are required to dissipate the voltage generated by this process. ClC-7, a Cl-/H+ antiporter responsible for lysosomal Cl- permeability, is a candidate to contribute to the acidification process as part of this 'counterion pathway'. The signaling lipid PI(3,5)P2 modulates lysosomal dynamics, including by regulating lysosomal ion channels, raising the possibility that it could contribute to lysosomal pH regulation. Here we demonstrate that depleting PI(3,5)P2 by inhibiting the PIKfyve kinase causes lysosomal hyperacidification, primarily via an effect on ClC-7. We further show that PI(3,5)P2 directly inhibits ClC-7 transport and that this inhibition is eliminated in a disease-causing gain-of-function ClC-7 mutation. Together these observations suggest an intimate role for ClC-7 in lysosomal pH regulation.
    Keywords:  human; molecular biophysics; structural biology
    DOI:  https://doi.org/10.7554/eLife.74136
  38. Chem Biol Interact. 2022 Jun 01. pii: S0009-2797(22)00208-3. [Epub ahead of print] 110003
    A pivotal role of selective autophagy in mitochondrial quality control: Implications for zinc oxide nanoparticles induced neurotoxicity.
    Liwei Wang, Zhiyu Duan, Madan Liang, Chunqiang Wang, Tingting Liang, Liqian Sun, Chaoqun Yan, Qingshan Li, Taigang Liang.
      Excessive occupational, medical, and environmental exposure of zinc oxide nanoparticles (ZnONPs) caused its accumulation in the nervous system and raised global concerns over its detrimental effects. However, very few researches had been conducted on the impact of mitochondrial quality control process on central nervous system (CNS) after ZnONPs administration, including mitochondrial fission, fusion, biogenesis, and autophagy. In present study, mitochondrial dysfunction and apoptosis were triggered in ZnONPs-exposed human neuroblastoma SH-SY5Y cells. Upregulation of mitochondrial biogenesis regulator (PGC-1α) and fission proteins (Drp1) and downregulation of fusion proteins (OPA1 and Mfn2) were observed in 3 and 6 μg/mL ZnONPs-treated cells. Meanwhile, loss of mitochondrial dynamics and biogenesis was observed in the severe impaired cells (treated with 12 μg/mL ZnONPs). More, autophagy and mitophagy were significantly activated in ZnONPs-treated cells. The increased Beclin1 and LC3 II proteins, decreases of p62 protein, and activated PINK1/Parkin signaling were quantified. The autophagy agonist (Rapamycin), inhibitor (3-MA), and mitophagy inhibitor (Cyclosporine A, CsA) were employed to verify the roles of autophagy and mitophagy in ZnONPs-treated cells. Consequently, mitochondrial dysfunction and apoptosis were aggravated by the blockage of autophagy and mitophagy. Our research could be used to evaluate the risk assessment of ZnONPs exposure in CNS neurons so as to provide a crucial guideline for their future biological applications.
    Keywords:  Autophagy; Mitochondrial quality control; Mitophagy; Neurotoxicity; Zinc oxide nanoparticles
    DOI:  https://doi.org/10.1016/j.cbi.2022.110003
  39. Sci Total Environ. 2022 Jun 02. pii: S0048-9697(22)03558-6. [Epub ahead of print] 156461
    Triphenyl phosphate (TPP) promotes hepatocyte toxicity via induction of endoplasmic reticulum stress and inhibition of autophagy flux.
    Miaoran Li, Gang Liu, Li-Xia Yuan, Jing Yang, Jing Liu, Zhijie Li, Chuanbin Yang, Jigang Wang.
      Triphenyl phosphate (TPP), a commonly used organophosphate flame retardant, is frequently found in environmental and biota samples, indicating widespread human exposure. Recent studies have shown that TPP causes hepatotoxicity, but the underlying cellular mechanisms are not fully elucidated. Here, by using normal hepatocyte AML12 cells as a model, we showed that TPP induced apoptotic cell death. RNA sequencing analyses revealed that differentially expressed genes induced by TPP were related to endoplasmic reticulum (ER) stress and autophagy. Immunostaining and western blot results further confirmed that TPP activated ER stress. Interestingly, though TPP increased LC3-II, a canonical marker for autophagy, TPP inhibited autophagy flux rather than induced autophagy. Interestingly, TPP-induced ER stress facilitated autophagy flux inhibition and apoptosis. Furthermore, inhibition of autophagy aggravated, and activation of autophagy attenuated apoptosis induced by TPP. Collectively, these results uncovered that ER stress and autophagy flux inhibition were responsible for TPP-induced apoptosis in mouse hepatocytes. Thus, our foundlings provided novel insight into the potential mechanisms of TPP-induced hepatocyte toxicity.
    Keywords:  Apoptosis; Autophagy; ER stress; Hepatoxicity; Triphenyl phosphate
    DOI:  https://doi.org/10.1016/j.scitotenv.2022.156461
  40. Int J Mol Sci. 2022 May 24. pii: 5894. [Epub ahead of print]23(11):
    The Common Cellular Events in the Neurodegenerative Diseases and the Associated Role of Endoplasmic Reticulum Stress.
    Soojeong Kim, Doo Kyung Kim, Seho Jeong, Jaemin Lee.
      Neurodegenerative diseases are inseparably linked with aging and increase as life expectancy extends. There are common dysfunctions in various cellular events shared among neurogenerative diseases, such as calcium dyshomeostasis, neuroinflammation, and age-associated decline in the autophagy-lysosome system. However, most of all, the prominent pathological feature of neurodegenerative diseases is the toxic buildup of misfolded protein aggregates and inclusion bodies accompanied by an impairment in proteostasis. Recent studies have suggested a close association between endoplasmic reticulum (ER) stress and neurodegenerative pathology in cellular and animal models as well as in human patients. The contribution of mutant or misfolded protein-triggered ER stress and its associated signaling events, such as unfolded protein response (UPR), to the pathophysiology of various neurodegenerative disorders, including Alzheimer's, Parkinson's, and Huntington's disease, amyotrophic lateral sclerosis, and prion disease, is described here. Impaired UPR action is commonly attributed to exacerbated ER stress, pathogenic protein aggregate accumulation, and deteriorating neurodegenerative pathologies. Thus, activating certain UPR components has been shown to alleviate ER stress and its associated neurodegeneration. However, uncontrolled activation of some UPR factors has also been demonstrated to worsen neurodegenerative phenotypes, suggesting that detailed molecular mechanisms around ER stress and its related neurodegenerations should be understood to develop effective therapeutics against aging-associated neurological syndromes. We also discuss current therapeutic endeavors, such as the development of small molecules that selectively target individual UPR components and address ER stress in general.
    Keywords:  Alzheimer’s disease; ER stress; Huntington’s disease; Parkinson’s disease; amyotrophic lateral sclerosis; endoplasmic reticulum; misfolded protein; neurodegenerative disease; prion disease; unfolded protein response
    DOI:  https://doi.org/10.3390/ijms23115894
  41. Nat Commun. 2022 Jun 07. 13(1): 3272
    Molecular basis of mEAK7-mediated human V-ATPase regulation.
    Rong Wang, Yu Qin, Xiao-Song Xie, Xiaochun Li.
      The activity of V-ATPase is well-known to be regulated by reversible dissociation of its V1 and Vo domains in response to growth factor stimulation, nutrient sensing, and cellular differentiation. The molecular basis of its regulation by an endogenous modulator without affecting V-ATPase assembly remains unclear. Here, we discover that a lysosome-anchored protein termed (mammalian Enhancer-of-Akt-1-7 (mEAK7)) binds to intact V-ATPase. We determine cryo-EM structure of human mEAK7 in complex with human V-ATPase in native lipid-containing nanodiscs. The structure reveals that the TLDc domain of mEAK7 engages with subunits A, B, and E, while its C-terminal domain binds to subunit D, presumably blocking V1-Vo torque transmission. Our functional studies suggest that mEAK7, which may act as a V-ATPase inhibitor, does not affect the activity of V-ATPase in vitro. However, overexpression of mEAK7 in HCT116 cells that stably express subunit a4 of V-ATPase represses the phosphorylation of ribosomal protein S6. Thus, this finding suggests that mEAK7 potentially links mTOR signaling with V-ATPase activity.
    DOI:  https://doi.org/10.1038/s41467-022-30899-z
  42. Dev Cell. 2022 Jun 06. pii: S1534-5807(22)00361-6. [Epub ahead of print]57(11): 1311-1313
    Major construction entails major demolition.
    Ekaterina Korotkevich, Takashi Hiiragi.
      Embryonic cells of the early mouse embryo become hypersensitive to apoptotic stimuli before gastrulation. In this issue of Developmental Cell, Pernaute et al. show that this switch in sensitivity is a result of a change in mitochondrial dynamics and mitophagy levels controlled by DRP1, a regulator of mitochondrial fission.
    DOI:  https://doi.org/10.1016/j.devcel.2022.05.008
  43. Neuropharmacology. 2022 Jun 01. pii: S0028-3908(22)00213-1. [Epub ahead of print]214 109154
    Knockdown of miRNA-134-5p rescues dendritic deficits by promoting AMPK-mediated mitophagy in a mouse model of depression.
    Guoli Wang, Ying Liu, Xuejie Zhu, Kehao Lin, Mingkai Li, Zhenke Wu, Ronghua Zhang, Qiusheng Zheng, Defang Li, Tianyue An.
      Neuronal dendrites and dendritic spines are essential for normal synaptic transmission and may be critically involved in the pathophysiology of various neurological disorders, including depression. Emerging data supports the role of mitochondria in dendritic protrusions in modulating the development and morphological plasticity of spines. Mitophagy, a mitochondria-specific form of autophagy, is the fundamental process of clearing damaged mitochondria to maintain cellular homeostasis. As a brain-specific microRNA, miR-134 is localized to the synaptodendritic compartment of hippocampal neurons and negatively regulates the development of dendritic spines. However, the role of miR-134 in mitophagy related to dendritic deficits in the pathophysiology of depression remains unclear. In this study, we showed that miR-134-5p knockdown abrogated depression-like behavioral symptoms and corrected aberrant spine morphology in hippocampal neurons of chronic unpredictable mild stress (CUMS) mice. Moreover, knockdown of miR-134-5p triggered autophagy in dendrites, improved mitochondrial impairment, and induced the generation of autophagosomes in the hippocampus of CUMS mice. We further found that AMP-activated protein kinase (AMPK), which mediates the impairment of defective mitochondria via mitophagy, can bind directly to miR-134-5p and is negatively regulated by this miRNA. This study demonstrates that miR-134-5p exerts an enormous effect on dendritic deficits by promoting AMPK-mediated mitophagy and provides a potential new target for antidepressant drug research and development.
    Keywords:  AMPK signaling Pathway; Dendrite; Depression; Mitophagy; miR-134-5p
    DOI:  https://doi.org/10.1016/j.neuropharm.2022.109154
  44. Am J Respir Crit Care Med. 2022 Jun 08.
    Extracellular Vesicles as Therapy for CDH-associated Pulmonary Hypoplasia: Extra! Extra! Read All About Autophagy!
    Matthew T Harting, David Warburton.
      
    Keywords:  autophagy; congenital diaphragmatic hernia; extracellular vesicles
    DOI:  https://doi.org/10.1164/rccm.202204-0635ED
  45. Aging Cell. 2022 Jun 05. e13615
    Central role for p62/SQSTM1 in the elimination of toxic tau species in a mouse model of tauopathy.
    Maiko Ono, Masaaki Komatsu, Bin Ji, Yuhei Takado, Masafumi Shimojo, Takeharu Minamihisamatsu, Eiji Warabi, Toru Yanagawa, Gen Matsumoto, Ichio Aoki, Nicholas M Kanaan, Tetsuya Suhara, Naruhiko Sahara, Makoto Higuchi.
      Intracellular accumulation of filamentous tau aggregates with progressive neuronal loss is a common characteristic of tauopathies. Although the neurodegenerative mechanism of tau-associated pathology remains unclear, molecular elements capable of degrading and/or sequestering neurotoxic tau species may suppress neurodegenerative progression. Here, we provide evidence that p62/SQSTM1, a ubiquitinated cargo receptor for selective autophagy, acts protectively against neuronal death and neuroinflammation provoked by abnormal tau accumulation. P301S mutant tau transgenic mice (line PS19) exhibited accumulation of neurofibrillary tangles with localization of p62 mostly in the brainstem, but neuronal loss with few neurofibrillary tangles in the hippocampus. In the hippocampus of PS19 mice, the p62 level was lower compared to the brainstem, and punctate accumulation of phosphorylated tau unaccompanied by co-localization of p62 was observed. In PS19 mice deficient in p62 (PS19/p62-KO), increased accumulation of phosphorylated tau, acceleration of neuronal loss, and exacerbation of neuroinflammation were observed in the hippocampus as compared with PS19 mice. In addition, increase of abnormal tau and neuroinflammation were observed in the brainstem of PS19/p62-KO. Immunostaining and dot-blot analysis with an antibody selectively recognizing tau dimers and higher-order oligomers revealed that oligomeric tau species in PS19/p62-KO mice were significantly accumulated as compared to PS19 mice, suggesting the requirement of p62 to eliminate disease-related oligomeric tau species. Our findings indicated that p62 exerts neuroprotection against tau pathologies by eliminating neurotoxic tau species, suggesting that the manipulative p62 and selective autophagy may provide an intrinsic therapy for the treatment of tauopathy.
    Keywords:  FTDP-17; autophagy; p62/SQSTM1; tau; tau oligomers; tauopathy model mouse
    DOI:  https://doi.org/10.1111/acel.13615
  46. Cells. 2022 Jun 04. pii: 1840. [Epub ahead of print]11(11):
    Lysosomal Proteomics Links Disturbances in Lipid Homeostasis and Sphingolipid Metabolism to CLN5 Disease.
    Stefano Doccini, Maria Marchese, Federica Morani, Nicola Gammaldi, Serena Mero, Francesco Pezzini, Rabah Soliymani, Melissa Santi, Giovanni Signore, Asahi Ogi, Silvia Rocchiccioli, Katja M Kanninen, Alessandro Simonati, Maciej M Lalowski, Filippo M Santorelli.
      CLN5 disease (MIM: 256731) represents a rare late-infantile form of neuronal ceroid lipofuscinosis (NCL), caused by mutations in the CLN5 gene that encodes the CLN5 protein (CLN5p), whose physiological roles stay unanswered. No cure is currently available for CLN5 patients and the opportunities for therapies are lagging. The role of lysosomes in the neuro-pathophysiology of CLN5 disease represents an important topic since lysosomal proteins are directly involved in the primary mechanisms of neuronal injury occurring in various NCL forms. We developed and implemented a lysosome-focused, label-free quantitative proteomics approach, followed by functional validations in both CLN5-knockout neuronal-like cell lines and Cln5-/- mice, to unravel affected pathways and modifying factors involved in this disease scenario. Our results revealed a key role of CLN5p in lipid homeostasis and sphingolipid metabolism and highlighted mutual NCL biomarkers scored with high lysosomal confidence. A newly generated cln5 knockdown zebrafish model recapitulated most of the pathological features seen in NCL disease. To translate the findings from in-vitro and preclinical models to patients, we evaluated whether two FDA-approved drugs promoting autophagy via TFEB activation or inhibition of the glucosylceramide synthase could modulate in-vitro ROS and lipid overproduction, as well as alter the locomotor phenotype in zebrafish. In summary, our data advance the general understanding of disease mechanisms and modifying factors in CLN5 disease, which are recurring in other NCL forms, also stimulating new pharmacological treatments.
    Keywords:  CLN5 disease; NCL; lysosomal proteomics; lysosomes; miglustat; trehalose
    DOI:  https://doi.org/10.3390/cells11111840
  47. Front Bioeng Biotechnol. 2022 ;10 849768
    An Overview of Autophagy in Hematopoietic Stem Cell Transplantation.
    Soheila Montazersaheb, Ali Ehsani, Ezzatollah Fathi, Raheleh Farahzadi, Ilja Vietor.
      Autophagy is a fundamental homeostatic process crucial for cellular adaptation in response to metabolic stress. Autophagy exerts its effect through degrading intracellular components and recycling them to produce macromolecular precursors and energy. This physiological process contributes to cellular development, maintenance of cellular/tissue homeostasis, immune system regulation, and human disease. Allogeneic hematopoietic stem cell transplantation (HSCT) is the only preferred therapy for most bone marrow-derived cancers. Unfortunately, HSCT can result in several serious and sometimes untreatable conditions due to graft-versus-host disease (GVHD), graft failure, and infection. These are the major cause of morbidity and mortality in patients receiving the transplant. During the last decade, autophagy has gained a considerable understanding of its role in various diseases and cellular processes. In light of recent research, it has been confirmed that autophagy plays a crucial role in the survival and function of hematopoietic stem cells (HSCs), T-cell differentiation, antigen presentation, and responsiveness to cytokine stimulation. Despite the importance of these events to HSCT, the role of autophagy in HSCT as a whole remains relatively ambiguous. As a result of the growing use of autophagy-modulating agents in the clinic, it is imperative to understand how autophagy functions in allogeneic HSCT. The purpose of this literature review is to elucidate the established and implicated roles of autophagy in HSCT, identifying this pathway as a potential therapeutic target for improving transplant outcomes.
    Keywords:  autophagy; cell therapy; cellular mechanism; hematopoietic stem cells; hematopoietic stem cells transplantation
    DOI:  https://doi.org/10.3389/fbioe.2022.849768
  48. Int J Mol Sci. 2022 May 28. pii: 6071. [Epub ahead of print]23(11):
    Effects of Individual Amino Acids on PPARα Transactivation, mTORC1 Activation, ApoA-I Transcription and pro-ApoA-I Secretion.
    Jehad Z Tayyeb, Herman E Popeijus, Janna van de Sanden, Willem Zwaan, Ronald P Mensink, Jogchum Plat.
      A higher concentration of apolipoprotein A-I (ApoA-I) is associated with increased high density lipoprotein functionality and reverse cholesterol transport (RCT). A promising strategy to prevent cardiovascular diseases is therefore to improve RCT by increasing de novo ApoA-I production. Since experimental animal models have suggested effects of amino acids on hepatic lipoprotein metabolism, we here examined the effects of different amino acids on hepatic ApoA-I production. Human hepatocytes (HepG2) were exposed to six individual amino acids for 48 h. ApoA-I transcription and secreted pro-ApoA-I protein concentrations were analyzed using quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assays (ELISA), respectively. Additionally, CPT1 and KEAP1 mRNA expression, peroxisome proliferator-activated receptor alpha (PPARα) transactivation, and mechanistic target of rapamycin complex 1 (mTORC1) phosphorylation were determined. Leucine, glutamic acid, and tryptophan increased ApoA-I and CPT1 mRNA expression. Tryptophan also strongly increased PPARα transactivation. Glutamine, proline, and histidine increased pro-ApoA-I protein concentrations but mTORC1 phosphorylation remained unchanged regardless of the amino acid provided. In conclusion, individual amino acids have different effects on ApoA-I mRNA expression and pro-ApoA-I production which can partially be explained by specific effects on PPARα transactivation, while mTORC1 phosphorylation remained unaffected.
    Keywords:  ApoA-I; PPARα; amino acids; mTOR
    DOI:  https://doi.org/10.3390/ijms23116071
  49. Oral Oncol. 2022 Jun 03. pii: S1368-8375(22)00237-8. [Epub ahead of print]131 105948
    Promising autophagy inhibitors: Therapeutic implications in oral cancer.
    K Shree Harini, Devaraj Ezhilarasan.
      
    DOI:  https://doi.org/10.1016/j.oraloncology.2022.105948
  50. Autophagy. 2022 Jun 06. 1-20
    Protective effect of the tunneling nanotube-TNFAIP2/M-sec system on podocyte autophagy in diabetic nephropathy.
    F Barutta, S Bellini, S Kimura, K Hase, B Corbetta, A Corbelli, F Fiordaliso, S Bruno, L Biancone, A Barreca, M G Papotti, E Hirsh, M Martini, R Gambino, M Durazzo, H Ohno, G Gruden.
      Podocyte injury leading to albuminuria is a characteristic feature of diabetic nephropathy (DN). Hyperglycemia and advanced glycation end products (AGEs) are major determinants of DN. However, the underlying mechanisms of podocyte injury remain poorly understood. The cytosolic protein TNFAIP2/M-Sec is required for tunneling nanotubes (TNTs) formation, which are membrane channels that transiently connect cells, allowing organelle transfer. Podocytes express TNFAIP2 and form TNTs, but the potential relevance of the TNFAIP2-TNT system in DN is unknown. We studied TNFAIP2 expression in both human and experimental DN and the renal effect of tnfaip2 deletion in streptozotocin-induced DN. Moreover, we explored the role of the TNFAIP2-TNT system in podocytes exposed to diabetes-related insults. TNFAIP2 was overexpressed by podocytes in both human and experimental DN and exposure of podocytes to high glucose and AGEs induced the TNFAIP2-TNT system. In diabetic mice, tnfaip2 deletion exacerbated albuminuria, renal function loss, podocyte injury, and mesangial expansion. Moreover, blockade of the autophagic flux due to lysosomal dysfunction was observed in diabetes-injured podocytes both in vitro and in vivo and exacerbated by tnfaip2 deletion. TNTs allowed autophagosome and lysosome exchange between podocytes, thereby ameliorating AGE-induced lysosomal dysfunction and apoptosis. This protective effect was abolished by tnfaip2 deletion, TNT inhibition, and donor cell lysosome damage. By contrast, Tnfaip2 overexpression enhanced TNT-mediated transfer and prevented AGE-induced autophagy and lysosome dysfunction and apoptosis. In conclusion, TNFAIP2 plays an important protective role in podocytes in the context of DN by allowing TNT-mediated autophagosome and lysosome exchange and may represent a novel druggable target.Abbreviations: AGEs: advanced glycation end products; AKT1: AKT serine/threonine kinase 1; AO: acridine orange; ALs: autolysosomes; APs: autophagosomes; BM: bone marrow; BSA: bovine serum albumin; CTSD: cathepsin D; DIC: differential interference contrast; DN: diabetic nephropathy; FSGS: focal segmental glomerulosclerosis; HG: high glucose; KO: knockout; LAMP1: lysosomal-associated membrane protein 1; LMP: lysosomal membrane permeabilization; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; PI3K: phosphoinositide 3-kinase; STZ: streptozotocin; TNF: tumor necrosis factor; TNFAIP2: tumor necrosis factor, alpha-induced protein 2; TNTs: tunneling nanotubes; WT: wild type.
    Keywords:  Advanced glycation end products; albuminuria; autophagosomes; experimental diabetes; hyperglycemia; lysosomes; nephrin; podocytes; renal function loss; slit diaphragm
    DOI:  https://doi.org/10.1080/15548627.2022.2080382
  51. Cell Death Dis. 2022 Jun 10. 13(6): 543
    Emerging role of LETM1/GRP78 axis in lung cancer.
    Quangdon Tran, Hyunji Lee, Jae Hun Jung, Seung-Hee Chang, Robin Shrestha, Gyeyeong Kong, Jisoo Park, Seon-Hwan Kim, Kyu-Sang Park, Hyun-Woo Rhee, Jeanho Yun, Myung-Haing Cho, Kwang Pyo Kim, Jongsun Park.
      The selective autophagy of damaged mitochondria is called mitophagy. Mitochondrial dysfunction, mitophagy, and apoptosis have been suggested to be interrelated in various human lung carcinomas. Leucine zipper EF-hand-containing transmembrane protein-1 (LETM1) was cloned in an attempt to identify candidate genes for Wolf-Hirschhorn syndrome. LETM1 plays a role in mitochondrial morphology, ion homeostasis, and cell viability. LETM1 has also been shown to be overexpressed in different human cancer tissues, including lung cancer. In the current study, we have provided clear evidence that LETM1 acts as an anchoring protein for the mitochondria-associated ER membrane (MAM). Fragmented mitochondria have been found in lung cancer cells with LETM1 overexpression. In addition, a reduction of mitochondrial membrane potential and significant accumulation of microtubule-associated protein 1 A/1B-light chain 3 punctate, which localizes with Red-Mito, was found in LETM1-overexpressed cells, suggesting that mitophagy is upregulated in these cells. Interestingly, glucose-regulated protein 78 kDa (GRP78; an ER chaperon protein) and glucose-regulated protein 75 kDa (GRP75) were posited to interact with LETM1 in the immunoprecipitated LETM1 of H460 cells. This interaction was enhanced in cells treated with carbonyl cyanide m-chlorophenylhydrazone, a chemical mitophagy inducer. Treatment of cells with honokiol (a GRP78 inhibitor) blocked LETM1-mediated mitophagy, and CRISPR/Cas9-mediated GRP75 knockout inhibited LETM1-induced autophagy. Thus, GRP78 interacts with LETM1. Taken together, these observations support the notion that the complex formation of LETM1/GRP75/GRP78 might be an important step in MAM formation and mitophagy, thus regulating mitochondrial quality control in lung cancer.
    DOI:  https://doi.org/10.1038/s41419-022-04993-5
  52. Neural Regen Res. 2022 Dec;17(12): 2606-2614
    From cradle to grave: neurogenesis, neuroregeneration and neurodegeneration in Alzheimer's and Parkinson's diseases.
    Debia Wakhloo, Jane Oberhauser, Angela Madira, Sameehan Mahajani.
      Two of the most common neurodegenerative disorders - Alzheimer's and Parkinson's diseases - are characterized by synaptic dysfunction and degeneration that culminate in neuronal loss due to abnormal protein accumulation. The intracellular aggregation of hyper-phosphorylated tau and the extracellular aggregation of amyloid beta plaques form the basis of Alzheimer's disease pathology. The major hallmark of Parkinson's disease is the loss of dopaminergic neurons in the substantia nigra pars compacta, following the formation of Lewy bodies, which consists primarily of alpha-synuclein aggregates. However, the discrete mechanisms that contribute to neurodegeneration in these disorders are still poorly understood. Both neuronal loss and impaired adult neurogenesis have been reported in animal models of these disorders. Yet these findings remain subject to frequent debate due to a lack of conclusive evidence in post mortem brain tissue from human patients. While some publications provide significant findings related to axonal regeneration in Alzheimer's and Parkinson's diseases, they also highlight the limitations and obstacles to the development of neuroregenerative therapies. In this review, we summarize in vitro and in vivo findings related to neurogenesis, neuroregeneration and neurodegeneration in the context of Alzheimer's and Parkinson's diseases.
    Keywords:  Tau; Wallerian degeneration; alpha-synuclein; amyloid beta plaques; autophagy; dopaminergic neurons; human iPSCs; mitochondrial dysfunction; scRNA sequencing; synaptic dysfunction
    DOI:  https://doi.org/10.4103/1673-5374.336138
  53. Neurosci Res. 2022 Jun 07. pii: S0168-0102(22)00172-9. [Epub ahead of print]
    BAX regulates dendritic spine development via mitochondrial fusion.
    Qinhua Gu, Kaizheng Duan, Ronald S Petralia, Ya-Xian Wang, Zheng Li.
      BAX is a Bcl-2 family protein acting on apoptosis. It also promotes mitochondrial fusion by interacting with the mitochondrial fusion protein Mitofusin (Mfn1 and Mfn2). Neuronal mitochondria are important for the development and modification of dendritic spines, which are subcellular compartments accommodating excitatory synapses in postsynaptic neurons. The abundance of dendritic mitochondria influences dendritic spine development. Mitochondrial fusion is essential for mitochondrial homeostasis. Here, we show that in the hippocampal neuron of BAX knockout mice, mitochondrial fusion is impaired, leading to decreases in mitochondrial length and total mitochondrial mass in dendrites. Notably, BAX knockout mice also have fewer dendritic spines and less cellular Adenosine 5'triphosphate (ATP) in dendrites. The spine and ATP changes are abolished by restoring mitochondria fusion via overexpressing Mfn1 and Mfn2. These findings indicate that BAX-mediated mitochondrial fusion in neurons is crucial for the development of dendritic spines and the maintenance of cellular ATP levels.
    Keywords:  ATP; Mfn; mitochondria; spine
    DOI:  https://doi.org/10.1016/j.neures.2022.06.002
  54. Cancer Treat Res Commun. 2022 Jun 03. pii: S2468-2942(22)00074-0. [Epub ahead of print]32 100584
    Evaluation of targeting autophagy for the treatment of malignant rhabdoid tumours.
    Patricia Hannon Barroeta, Stefania Magnano, Maureen J O'Sullivan, Daniela M Zisterer.
      Malignant rhabdoid tumour (MRT) is a rare and aggressive paediatric tumour that typically arises in the kidneys or central nervous system (CNS). The malignancy often affects patients under the age of three and is associated with an extremely poor survival rate, with most deaths occurring within the first year of presentation. Thus, there is an unmet and urgent medical need for novel therapeutic strategies for this malignancy. One of the major issues when treating MRT patients is the emergence of chemoresistance. Autophagy has become an area of focus in the study of chemoresistance due to its reported dual role as both a pro-survival and pro-death mechanism. The role of autophagy in the chemotherapeutic response of MRT remains largely unknown. A greater understanding of the role of autophagy may lead to the development of therapeutic strategies to enhance chemotherapeutic effect and improve the clinical outcome of MRT patients. This study evaluated the cellular response to cisplatin, a representative chemotherapeutic agent used in the treatment of MRT, and the role of autophagy in mediating cisplatin resistance. Our results demonstrated that cisplatin induced apoptosis and autophagy concomitantly in a panel of MRT cell lines. Furthermore, inhibition of caspase-induced apoptosis with Z-VAD-FMK also inhibited autophagy levels demonstrating a complex interplay between these two pathways. In addition, blocking autophagy at the early stages of the autophagic process using the pharmacological inhibitor SAR405 or through the genetic knockdown of critical autophagic protein ATG5 by siRNA did not sensitise cells to cisplatin-induced apoptosis. Collectively, these results suggest that induction of autophagy does not appear to elicit a pro-survival effect in the chemotherapeutic response of MRT cells.
    Keywords:  Apoptosis; Autophagy; Chemoresistance; Malignant rhabdoid tumour
    DOI:  https://doi.org/10.1016/j.ctarc.2022.100584
  55. Front Vet Sci. 2022 ;9 818294
    ERAS Is Constitutively Expressed in the Tissues of Adult Horses and May Be a Key Player in Basal Autophagy.
    Francesca De Falco, Antonella Perillo, Fabio Del Piero, Chiara Del Prete, Nicola Zizzo, Ioan Marcus, Sante Roperto.
      ERas is a new gene of the Ras family found in murine embryonic stem (ES) cells. Its human ortholog is not expressed in human ES cells. So far ERas gene has only been found to be expressed in the tissues of adult cynomolgus monkeys and cattle; however, information about ERAS expression or its potential functions in equine tissues is lacking. This study was performed to investigate whether Eras is an equine functional gene and whether ERAS is expressed in the tissues of adult horses and determine its potential physiological role. Expression of the ERas gene was detected in all examined adult tissues, and the RT-PCR assay revealed ERAS transcripts. Protein expression was also detected by Western blot analysis. Quantitative real time RT-qPCR analysis revealed that different expression levels of ERAS transcripts were most highly expressed in the testis. Immunohistochemically, ERAS was found to be localized prevalently in the plasmatic membrane as well as cytoplasm of the cells. ERAS was a physical partner of activated PDGFβR leading to the AKT signaling. ERAS was found to interact with a network of proteins (BAG3, CHIP, Hsc70/Hsp70, HspB8, Synpo2, and p62) known to play a role in the chaperone-assisted selective autophagy (CASA), which is also known as BAG3-mediated selective macroautophagy, an adaptive mechanism to maintain cellular homeostasis. Furthermore, ERAS was found to interact with parkin. PINK1, BNIP3, laforin. All these proteins are known to play a role in parkin-dependent and -independent mitophagy. This is the first study demonstrating that Eras is a functional gene, and that ERAS is constitutively expressed in the tissues of adult horses. ERAS appears to play a physiological role in cellular proteostasis maintenance, thus mitigating the proteotoxicity of accumulated misfolded proteins and contributing to protection against disease. Finally, it is conceivable that activation of AKT pathway by PDGFRs promotes actin reorganization, directed cell movements, stimulation of cell growth.
    Keywords:  ERas; adult horse; basal autophagy; constitutive expression; mitophagy
    DOI:  https://doi.org/10.3389/fvets.2022.818294
  56. Cureus. 2022 May;14(5): e24803
    An Update to Hallmarks of Cancer.
    Swapna Ravi, Antonio M Alencar, Jemma Arakelyan, Weihao Xu, Roberta Stauber, Cheng-Chi I Wang, Ruzanna Papyan, Narine Ghazaryan, Rosalina M Pereira.
      In the last decade, there has been remarkable progress in research toward understanding and refining the hallmarks of cancer. In this review, we propose a new hallmark - "pro-survival autophagy." The importance of pro-survival autophagy is well established in tumorigenesis, as it is related to multiple steps in cancer progression and vital for some cancers. Autophagy is a potential anti-cancer therapeutic target. For this reason, autophagy is a good candidate as a new hallmark of cancer. We describe two enabling characteristics that play a major role in enabling cells to acquire the hallmarks of cancer - "tumor-promoting microenvironment and macroenvironment" and "cancer epigenetics, genome instability and mutation." We also discuss the recent updates, therapeutic and prognostic implications of the eight hallmarks of cancer described by Hanahan et al. in 2011. Understanding these hallmarks and enabling characteristics is key not only to developing new ways to treat cancer efficiently but also to exploring options to overcome cancer resistance to treatment.
    Keywords:  autophagy; cancer; genome instability; hallmark; macroenvironment; microenvironment; tumor
    DOI:  https://doi.org/10.7759/cureus.24803
  57. Cells. 2022 May 31. pii: 1811. [Epub ahead of print]11(11):
    Endolysosome Iron Chelation Inhibits HIV-1 Protein-Induced Endolysosome De-Acidification-Induced Increases in Mitochondrial Fragmentation, Mitophagy, and Cell Death.
    Peter W Halcrow, Nirmal Kumar, Darius N K Quansah, Aparajita Baral, Braelyn Liang, Jonathan D Geiger.
      People with human immunodeficiency virus-1 (PLWH) experience high rates of HIV-1-associated neurocognitive disorders (HANDs); clinical symptoms range from being asymptomatic to experiencing HIV-associated dementia. Antiretroviral therapies have effectively prolonged the life expectancy related to PLWH; however, the prevalence of HANDs has increased. Implicated in the pathogenesis of HANDs are two HIV-1 proteins, transactivator of transcription (Tat) and gp120; both are neurotoxic and damage mitochondria. The thread-like morphological features of functional mitochondria become fragmented when levels of reactive oxygen species (ROS) increase, and ROS can be generated via Fenton-like chemistry in the presence of ferrous iron (Fe2+). Endolysosomes are central to iron trafficking in cells and contain readily releasable Fe2+ stores. However, it is unclear whether the endolysosome store is sufficient to account for insult-induced increases in levels of ROS, mitochondrial fragmentation, autophagy, and cell death. Using U87MG astrocytoma and SH-SY5Y neuroblastoma cells, we determined that chloroquine (CQ), Tat, and gp120 all (1) de-acidified endolysosomes, (2) decreased endolysosome numbers and increased endolysosome sizes, (3) increased mitochondrial numbers (fragmentation), (4) increased autophagosome numbers, (5) increased autolysosome numbers, (6) increased mitochondrial fragments within endolysosomes, and (7) increased cell death. These effects were all blocked by the endolysosome-specific iron chelator deferoxamine (DFO). Thus, the endolysosome de-acidification-induced release of endolysosome Fe2+ is sufficient to account for inter-organellar signaling events and cell biology consequences of HIV-1 proteins, including mitochondrial fragmentation, autophagy, and cell death.
    Keywords:  autophagosomes; autophagy; deferoxamine; endolysosomes; mitochondria; mitophagy; pH
    DOI:  https://doi.org/10.3390/cells11111811
  58. Sci Transl Med. 2022 Jun 08. 14(648): eabj2658
    Preclinical and clinical evaluation of the LRRK2 inhibitor DNL201 for Parkinson's disease.
    Danna Jennings, Sarah Huntwork-Rodriguez, Anastasia G Henry, Jennifer C Sasaki, René Meisner, Dolores Diaz, Hilda Solanoy, Xiang Wang, Elvira Negrou, Vitaliy V Bondar, Rajarshi Ghosh, Michael T Maloney, Nicholas E Propson, Yuda Zhu, Romeo D Maciuca, Laura Harris, Angela Kay, Peter LeWitt, T Alex King, Drew Kern, Aaron Ellenbogen, Ira Goodman, Andrew Siderowf, Jason Aldred, Omid Omidvar, Shababa T Masoud, Sonnet S Davis, Annie Arguello, Anthony A Estrada, Javier de Vicente, Zachary K Sweeney, Giuseppe Astarita, Marie T Borin, Bradley K Wong, Harvey Wong, Hoang Nguyen, Kimberly Scearce-Levie, Carole Ho, Matthew D Troyer.
      Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic risk factors for Parkinson's disease (PD). Increased LRRK2 kinase activity is thought to impair lysosomal function and may contribute to the pathogenesis of PD. Thus, inhibition of LRRK2 is a potential disease-modifying therapeutic strategy for PD. DNL201 is an investigational, first-in-class, CNS-penetrant, selective, ATP-competitive, small-molecule LRRK2 kinase inhibitor. In preclinical models, DNL201 inhibited LRRK2 kinase activity as evidenced by reduced phosphorylation of both LRRK2 at serine-935 (pS935) and Rab10 at threonine-73 (pT73), a direct substrate of LRRK2. Inhibition of LRRK2 by DNL201 demonstrated improved lysosomal function in cellular models of disease, including primary mouse astrocytes and fibroblasts from patients with Gaucher disease. Chronic administration of DNL201 to cynomolgus macaques at pharmacologically relevant doses was not associated with adverse findings. In phase 1 and phase 1b clinical trials in 122 healthy volunteers and in 28 patients with PD, respectively, DNL201 at single and multiple doses inhibited LRRK2 and was well tolerated at doses demonstrating LRRK2 pathway engagement and alteration of downstream lysosomal biomarkers. Robust cerebrospinal fluid penetration of DNL201 was observed in both healthy volunteers and patients with PD. These data support the hypothesis that LRRK2 inhibition has the potential to correct lysosomal dysfunction in patients with PD at doses that are generally safe and well tolerated, warranting further clinical development of LRRK2 inhibitors as a therapeutic modality for PD.
    DOI:  https://doi.org/10.1126/scitranslmed.abj2658
  59. PLoS Pathog. 2022 Jun 10. 18(6): e1010089
    Stress- and metabolic responses of Candida albicans require Tor1 kinase N-terminal HEAT repeats.
    Wanjun Qi, Maikel Acosta-Zaldivar, Peter R Flanagan, Ning-Ning Liu, Niketa Jani, José F Fierro, María T Andrés, Gary P Moran, Julia R Köhler.
      Whether to commit limited cellular resources toward growth and proliferation, or toward survival and stress responses, is an essential determination made by Target of Rapamycin Complex 1 (TORC1) for a eukaryotic cell in response to favorable or adverse conditions. Loss of TORC1 function is lethal. The TORC1 inhibitor rapamycin that targets the highly conserved Tor kinase domain kills fungal pathogens like Candida albicans, but is also severely toxic to human cells. The least conserved region of fungal and human Tor kinases are the N-terminal HEAT domains. We examined the role of the 8 most N-terminal HEAT repeats of C. albicans Tor1. We compared nutritional- and stress responses of cells that express a message for N-terminally truncated Tor1 from repressible tetO, with cells expressing wild type TOR1 from tetO or from the native promoter. Some but not all stress responses were significantly impaired by loss of Tor1 N-terminal HEAT repeats, including those to oxidative-, cell wall-, and heat stress; in contrast, plasma membrane stress and antifungal agents that disrupt plasma membrane function were tolerated by cells lacking this Tor1 region. Translation was inappropriately upregulated during oxidative stress in cells lacking N-terminal Tor1 HEAT repeats despite simultaneously elevated Gcn2 activity, while activation of the oxidative stress response MAP kinase Hog1 was weak. Conversely, these cells were unable to take advantage of favorable nutritional conditions by accelerating their growth. Consuming oxygen more slowly than cells containing wild type TOR1 alleles during growth in glucose, cells lacking N-terminal Tor1 HEAT repeats additionally were incapable of utilizing non-fermentable carbon sources. They were also hypersensitive to inhibitors of specific complexes within the respiratory electron transport chain, suggesting that inefficient ATP generation and a resulting dearth of nucleotide sugar building blocks for cell wall polysaccharides causes cell wall integrity defects in these mutants. Genome-wide expression analysis of cells lacking N-terminal HEAT repeats showed dysregulation of carbon metabolism, cell wall biosynthetic enzymes, translational machinery biosynthesis, oxidative stress responses, and hyphal- as well as white-opaque cell type-associated genes. Targeting fungal-specific Tor1 N-terminal HEAT repeats with small molecules might selectively abrogate fungal viability, especially when during infection multiple stresses are imposed by the host immune system.
    DOI:  https://doi.org/10.1371/journal.ppat.1010089
  60. Cell Mol Life Sci. 2022 Jun 09. 79(7): 354
    The protective effect of selenoprotein M on non-alcoholic fatty liver disease: the role of the AMPKα1-MFN2 pathway and Parkin mitophagy.
    Jingzeng Cai, Jiaqiang Huang, Jie Yang, Xiaoming Chen, Haoran Zhang, Yue Zhu, Qi Liu, Ziwei Zhang.
      Non-alcoholic fatty liver disease (NAFLD) is related to a dysregulation of mitophagy, a process that is not fully understood. Parkin-related mitophagy can sustain mitochondrial homeostasis and hepatocyte viability. Herein, we report that selenoprotein M (SELENOM) plays a central role in maintaining mitophagy in high-fat diet (HFD)-mediated NAFLD. We show that SELENOM was significantly downregulated in the liver of HFD-fed mice. SELENOM deletion aggravated HFD-mediated hepatic steatosis, inflammation, and fibrosis; accompanied by enhanced fatty acid oxidation and oxidative stress in the liver. Molecular analyses show that lipotoxicity was related to increased mitochondrial apoptosis as evidenced by enhanced mitochondrial ROS production, and attenuation of mitochondrial potential in the liver of HFD-fed SELENOM-/- mice. Additionally, SELENOM deletion reduced mitophagy and aggravated hepatic injury in NAFLD. Mechanistically, SELENOM overexpression activated Parkin-mediated mitophagy to reduce mitochondrial apoptosis and remove HFD-damaged mitochondria. We further found that SELENOM regulates Parkin expression via the AMPKα1-MFN2 pathway; blockade of AMPKα1 prevented SELENOM activation of Parkin-mediated mitophagy. Our work identified SELENOM downregulation as a possible explanation for the defective mitophagy in NAFLD. Thus, targeting SELENOM may be potential new therapeutic modalities for NAFLD treatment.
    Keywords:  Mitochondria homeostasis; Mitophagy; NAFLD; Oxidative stress; SELENOM
    DOI:  https://doi.org/10.1007/s00018-022-04385-0
  61. Cancer Med. 2022 Jun 09.
    Clustering analysis revealed the autophagy classification and potential autophagy regulators' sensitivity of pancreatic cancer based on multi-omics data.
    Yonghao Chen, Jialin Meng, Xiaofan Lu, Xiao Li, Chunhui Wang.
       BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy and is unresponsive to conventional therapeutic modalities due to its high heterogeneity, expounding the necessity, and priority of searching for effective biomarkers and drugs. Autophagy, as an evolutionarily conserved biological process, is upregulated in PDAC and its regulation is linked to a poor prognosis. Increased autophagy sequestered MHC-I on PDAC cells and weaken the antigen presentation and antitumor immune response, indicating the potential therapeutic strategies of autophagy inhibitors.
    METHODS: By performing 10 state-of-the-art multi-omics clustering algorithms, we constructed a robust PDAC classification model to reveal the autophagy-related genes among different subgroups.
    OUTCOMES: After building a more comprehensive regulating network for potential autophagy regulators exploration, we concluded the top 20 autophagy-related hub genes (GAPDH, MAPK3, RHEB, SQSTM1, EIF2S1, RAB5A, CTSD, MAP1LC3B, RAB7A, RAB11A, FADD, CFKN2A, HSP90AB1, VEGFA, RELA, DDIT3, HSPA5, BCL2L1, BAG3, and ERBB2), six miRNAs, five transcription factors, and five immune infiltrated cells as biomarkers. The drug sensitivity database was screened based on the biomarkers to predict possible drug-targeting signal pathways, hoping to yield novel insights, and promote the progress of the anticancer therapeutic strategy.
    CONCLUSION: We succefully constructed an autophagy-related mRNA/miRNA/TF/Immune cells network based on a 10 state-of art algorithm multi-omics analysis, and screened the drug sensitivity dataset for detecting potential signal pathway which might be possible autophagy modulators' targets.
    Keywords:  autophagy; bioinformation; drug sensitivity; multi-omics; pancreatic carcinoma
    DOI:  https://doi.org/10.1002/cam4.4932
  62. Front Aging Neurosci. 2022 ;14 878303
    Autophagy Impairment in App Knock-in Alzheimer's Model Mice.
    Richeng Jiang, Makoto Shimozawa, Johanna Mayer, Simone Tambaro, Rakesh Kumar, Axel Abelein, Bengt Winblad, Nenad Bogdanovic, Per Nilsson.
      Alzheimer's disease (AD) is characterized by impaired protein homeostasis leading to amyloid-β peptide (Aβ) amyloidosis. Amyloid precursor protein (APP) knock-in mice exhibit robust Aβ pathology, providing possibilities to determine its effect on protein homeostasis including autophagy. Here we compared human AD postmortem brain tissue with brains from two different types of App knock-in mice, App NL-F and App NL-G-F mice, exhibiting AD-like pathology. In AD postmortem brains, p62 levels are increased and p62-positive staining is detected in neurons, including potential axonal beadings, as well as in the vasculature and in corpora amylacea. Interestingly, p62 is also increased in the neurons in 12-month-old App NL-G-F mice. In brain homogenates from 12-month-old App NL-G-F mice, both p62 and light chain 3 (LC3)-II levels are increased as compared to wildtype (WT) mice, indicating inhibited autophagy. Double immunostaining for LC3 and Aβ revealed LC3-positive puncta in hippocampus of 24-month-old App NL-F mice around the Aβ plaques which was subsequently identified by electron microscopy imaging as an accumulation of autophagic vacuoles in dystrophic neurites around the Aβ plaques. Taken together, autophagy is impaired in App knock-in mice upon increased Aβ pathology, indicating that App knock-in mouse models provide a platform for understanding the correlation between Aβ and autophagy.
    Keywords:  APP knock-in mice; Alzheimer’s disease; LC3; amyloid beta; autophagy; electron microscopy; p62; protein homeostasis
    DOI:  https://doi.org/10.3389/fnagi.2022.878303
  63. Int J Mol Sci. 2022 Jun 01. pii: 6208. [Epub ahead of print]23(11):
    The Role of Autophagy and Pyroptosis in Liver Disorders.
    Huijie Zhao, Huiyang Liu, Yihan Yang, Honggang Wang.
      Pyroptosis is a programmed cell death caused by inflammasomes, which can detect cell cytosolic contamination or disturbance. In pyroptosis, caspase-1 or caspase-11/4/5 is activated, cleaving gasdermin D to separate its N-terminal pore-forming domain (PFD). The oligomerization of PFD forms macropores in the membrane, resulting in swelling and membrane rupture. According to the different mechanisms, pyroptosis can be divided into three types: canonical pathway-mediated pyroptosis, non-canonical pathway-mediated pyroptosis, and caspase-3-induced pyroptosis. Pyroptosis has been reported to play an important role in many tissues and organs, including the liver. Autophagy is a highly conserved process of the eukaryotic cell cycle. It plays an important role in cell survival and maintenance by degrading organelles, proteins and macromolecules in the cytoplasm. Therefore, the dysfunction of this process is involved in a variety of pathological processes. In recent years, autophagy and pyroptosis and their interactions have been proven to play an important role in various physiological and pathological processes, and have gradually attracted more and more attention to become a research hotspot. Therefore, this review summarized the role of autophagy and pyroptosis in liver disorders, and analyzed the related mechanism to provide a basis for future research.
    Keywords:  autophagy; hepatocellular carcinoma; hepatotoxicity; non-alcoholic fatty liver disease; pyroptosis
    DOI:  https://doi.org/10.3390/ijms23116208
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