bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2022–07–24
fifty-two papers selected by
Viktor Korolchuk, Newcastle University
  1. An overview of the molecular mechanisms of mitophagy in yeast.
  2. Sestrin mediates detection of and adaptation to low-leucine diets in Drosophila.
  3. Multiple functions of CALCOCO family proteins in selective autophagy.
  4. Glutamine Availability Regulates the Development of Aging Mediated by mTOR Signaling and Autophagy.
  5. Crosstalk of organelles in Parkinson's disease - MiT family transcription factors as central players in signaling pathways connecting mitochondria and lysosomes.
  6. Autophagy in health and disease: From molecular mechanisms to therapeutic target.
  7. Pharmacological modulation of autophagy for Alzheimer's disease therapy: Opportunities and obstacles.
  8. Atg23 is a vesicle-tethering protein.
  9. Cellular and mitochondrial quality control mechanisms in maintaining homeostasis in ageing.
  10. Targeting retinoic acid receptor alpha-corepressor interaction activates chaperone-mediated autophagy and protects against retinal degeneration.
  11. Induction of Autophagy Promotes Clearance of RHOP23H Aggregates and Protects From Retinal Degeneration.
  12. TBK1 is part of a galectin 8 dependent membrane damage recognition complex and drives autophagy upon Adenovirus endosomal escape.
  13. Development of an RHEB-Targeting Peptide To Inhibit mTORC1 Kinase Activity.
  14. Autophagy Activation Induces p62-Dependent Autophagic Degradation of Dengue Virus Capsid Protein During Infection.
  15. Stimulation of synaptic activity promotes TFEB-mediated clearance of pathological MAPT/Tau in cellular and mouse models of tauopathies.
  16. Confirmation of Decreased Rates of Cerebral Protein Synthesis In Vivo in a Mouse Model of Tuberous Sclerosis Complex.
  17. An ER-targeting DNA Nanodevice for Autophagy-dependent Degradation of Proteins in Membrane-bound Organelles.
  18. TFEB induces mitochondrial itaconate synthesis to suppress bacterial growth in macrophages.
  19. Frustration analysis of TBK1 missense mutations reported in ALS/FTD and cancer patients.
  20. Integrated regulation of stress responses, autophagy and survival by altered intracellular iron stores.
  21. Celastrol enhances transcription factor EB (TFEB)-mediated autophagy and mitigates Tau pathology: Implications for Alzheimer's disease therapy.
  22. MANF Inhibits α-Synuclein Accumulation through Activation of Autophagic Pathways.
  23. Compounds activating VCP D1 ATPase enhance both autophagic and proteasomal neurotoxic protein clearance.
  24. Transcription factor EB-mediated mesenchymal stem cell therapy induces autophagy and alleviates spinocerebellar ataxia type 3 defects in neuronal cells model.
  25. Analysis of ER-Phagy in Cancer Drug Resistance.
  26. Lack of COL6/collagen VI causes megakaryocyte dysfunction by impairing autophagy and inducing apoptosis.
  27. Nutritional strategies for autophagy activation and health consequences of autophagy impairment.
  28. Interaction Between Autophagy and Porphyromonas gingivalis-Induced Inflammation.
  29. Pathological mitophagy disrupts mitochondrial homeostasis in Leber's hereditary optic neuropathy.
  30. Rest and Digest-The Basal Role of Autophagy in Neurons and Its Relevance to Parkinson's Disease.
  31. Repurposing econazole as a pharmacological autophagy inhibitor to treat pancreatic ductal adenocarcinoma.
  32. Hepatocyte β-Catenin Loss is Compensated by Insulin-mTORC1 Activation to Promote Liver Regeneration.
  33. Autophagic membranes participate in hepatitis B virus nucleocapsid assembly, precore and core protein trafficking, and viral release.
  34. A small molecule Toll-like receptor antagonist rescues α-synuclein fibril pathology.
  35. mTOR contributes to endothelium-dependent vasorelaxation by promoting eNOS expression and preventing eNOS uncoupling.
  36. Activating mTOR mutations are detrimental in nutrient-poor conditions.
  37. Super-resolution analyzing spatial organization of lysosomes with an organic fluorescent probe.
  38. Reversible mitophagy drives metabolic suppression in diapausing beetles.
  39. A CRISPR screen targeting PI3K effectors identifies RASA3 as a negative regulator of LFA-1-mediated adhesion in T cells.
  40. Alzheimer's disease: Ablating single master site abolishes tau hyperphosphorylation.
  41. The Rab GTPase in the heart: Pivotal roles in development and disease.
  42. The interplay of epilepsy with impaired mitophagy and autophagy linked dementia (MAD): A review of therapeutic approaches.
  43. FUNDC1-mediated mitophagy and HIF1α activation drives pulmonary hypertension during hypoxia.
  44. MT-12 inhibits the proliferation of bladder cells in vitro and in vivo by enhancing autophagy through mitochondrial dysfunction.
  45. High glucose and palmitic acid induces neuronal senescence by NRSF/REST elevation and the subsequent mTOR-related autophagy suppression.
  46. Autophagy in Bone Remodeling: A Regulator of Oxidative Stress.
  47. Atorvastatin ameliorates doxorubicin-induced cardiomyopathy by regulating the autophagy-lysosome pathway and its upstream regulatory factor transcription factor EB.
  48. RAC3 Inhibition Induces Autophagy to Impair Metastasis in Bladder Cancer Cells via the PI3K/AKT/mTOR Pathway.
  49. Mitochondrial function and nutrient sensing pathways in ageing: enhancing longevity through dietary interventions.
  50. Neuronal activity induces glucosylceramide that is secreted via exosomes for lysosomal degradation in glia.
  51. ADAR1 downregulation by autophagy drives senescence independently of RNA editing by enhancing p16INK4a levels.
  52. The kidney-expressed transcription factor ZKSCAN3 is dispensable for autophagy transcriptional regulation and AKI progression in mouse.
  1. Biochim Biophys Acta Gen Subj. 2022 Jul 13. pii: S0304-4165(22)00121-0. [Epub ahead of print] 130203
    An overview of the molecular mechanisms of mitophagy in yeast.
    Ramona Schuster, Koji Okamoto.
      Autophagy-dependent selective degradation of excess or damaged mitochondria, termed mitophagy, is a tightly regulated process necessary for mitochondrial quality and quantity control. Mitochondria are highly dynamic and major sites for vital cellular processes such as ATP and iron‑sulfur cluster biogenesis. Due to their pivotal roles for immunity, apoptosis, and aging, the maintenance of mitochondrial function is of utmost importance for cellular homeostasis. In yeast, mitophagy is mediated by the receptor protein Atg32 that is localized to the outer mitochondrial membrane. Upon mitophagy induction, Atg32 expression is transcriptionally upregulated, which leads to its accumulation on the mitochondrial surface and to recruitment of the autophagic machinery via its direct interaction with Atg11 and Atg8. Importantly, post-translational modifications such as phosphorylation further fine-tune the mitophagic response. This review summarizes the current knowledge about mitophagy in yeast and its connection with mitochondrial dynamics and the ubiquitin-proteasome system.
    Keywords:  Atg11; Atg32; Atg8; Autophagy; Mitochondria; Yeast
    DOI:  https://doi.org/10.1016/j.bbagen.2022.130203
  2. Nature. 2022 Jul 20.
    Sestrin mediates detection of and adaptation to low-leucine diets in Drosophila.
    Xin Gu, Patrick Jouandin, Pranav V Lalgudi, Rich Binari, Max L Valenstein, Michael A Reid, Annamarie E Allen, Nolan Kamitaki, Jason W Locasale, Norbert Perrimon, David M Sabatini.
      Mechanistic target of rapamycin complex 1 (mTORC1) regulates cell growth and metabolism in response to multiple nutrients, including the essential amino acid leucine1. Recent work in cultured mammalian cells established the Sestrins as leucine-binding proteins that inhibit mTORC1 signalling during leucine deprivation2,3, but their role in the organismal response to dietary leucine remains elusive. Here we find that Sestrin-null flies (Sesn-/-) fail to inhibit mTORC1 or activate autophagy after acute leucine starvation and have impaired development and a shortened lifespan on a low-leucine diet. Knock-in flies expressing a leucine-binding-deficient Sestrin mutant (SesnL431E) have reduced, leucine-insensitive mTORC1 activity. Notably, we find that flies can discriminate between food with or without leucine, and preferentially feed and lay progeny on leucine-containing food. This preference depends on Sestrin and its capacity to bind leucine. Leucine regulates mTORC1 activity in glial cells, and knockdown of Sesn in these cells reduces the ability of flies to detect leucine-free food. Thus, nutrient sensing by mTORC1 is necessary for flies not only to adapt to, but also to detect, a diet deficient in an essential nutrient.
    DOI:  https://doi.org/10.1038/s41586-022-04960-2
  3. J Cell Physiol. 2022 Jul 19.
    Multiple functions of CALCOCO family proteins in selective autophagy.
    Wei Chen, Xueqian Ouyang, Linxi Chen, Lanfang Li.
      Selective autophagy is the lysosomal degradation of specific intracellular components sequestered into autophagosomes, late endosomes, or lysosomes through the activity of selective autophagy receptors. CALCOCO family proteins are the newly found selective autophagy receptors, which include calcium binding and coiled-coil domain 1 (CALCOCO1), calcium binding and coiled-coil domain 2/nuclear domain 10 protein 52 (CALCOCO2/NDP52), and calcium binding and coiled-coil domain 3/Tax1-binding protein 1 (CALCOCO3/TAX1BP1). Specifically, CALCOCO1 can be recruited to endoplasmic reticulum (ER) and Golgi to mediate selective ER-phagy and Golgiphagy. CALCOCO2 and CALCOCO3, which are two essential cargo receptors, can mediate mitophagy and xenophagy through interacting with autophagy-related-8/microtubule-associated protein 1 light chain 3 (ATG8/LC3) on the growing autophagosome, and binding ubiquitin for cargo recruitment. Considering the significance of these proteins in selective autophagy, we review the structures, distribution, posttranslational modifications, and phylogenetic analysis of CALCOCO family proteins and their roles in different selective autophagy.
    Keywords:  CALCOCO1; CALCOCO2; CALCOCO3; ER-phagy; mitophagy; selective autophagy
    DOI:  https://doi.org/10.1002/jcp.30836
  4. Front Pharmacol. 2022 ;13 924081
    Glutamine Availability Regulates the Development of Aging Mediated by mTOR Signaling and Autophagy.
    Jiao Zhou, Honghan Chen, Jintao Du, Haoran Tai, Xiaojuan Han, Ning Huang, Xiaobo Wang, Hui Gong, Mingyao Yang, Hengyi Xiao.
      Glutamine is a conditionally essential amino acid involved in energy production and redox homeostasis. Aging is commonly characterized by energy generation reduction and redox homeostasis dysfunction. Various aging-related diseases have been reported to be accompanied by glutamine exhaustion. Glutamine supplementation has been used as a nutritional therapy for patients and the elderly, although the mechanism by which glutamine availability affects aging remains elusive. Here, we show that chronic glutamine deprivation induces senescence in fibroblasts and aging in Drosophila melanogaster, while glutamine supplementation protects against oxidative stress-induced cellular senescence and rescues the D-galactose-prompted progeria phenotype in mice. Intriguingly, we found that long-term glutamine deprivation activates the Akt-mTOR pathway, together with the suppression of autolysosome function. However, the inhibition of the Akt-mTOR pathway effectively rescued the autophagy impairment and cellular senescence caused by glutamine deprivation. Collectively, our study demonstrates a novel interplay between glutamine availability and the aging process. Mechanistically, long-term glutamine deprivation could evoke mammalian target of rapamycin (mTOR) pathway activation and autophagy impairment. These findings provide new insights into the connection between glutamine availability and the aging process.
    Keywords:  aging; autophagy; cellular senescence; glutamine; mTOR
    DOI:  https://doi.org/10.3389/fphar.2022.924081
  5. Mol Neurodegener. 2022 Jul 16. 17(1): 50
    Crosstalk of organelles in Parkinson's disease - MiT family transcription factors as central players in signaling pathways connecting mitochondria and lysosomes.
    Martin Lang, Peter P Pramstaller, Irene Pichler.
      Living organisms constantly need to adapt to their surrounding environment and have evolved sophisticated mechanisms to deal with stress. Mitochondria and lysosomes are central organelles in the response to energy and nutrient availability within a cell and act through interconnected mechanisms. However, when such processes become overwhelmed, it can lead to pathologies. Parkinson's disease (PD) is a common neurodegenerative disorder (NDD) characterized by proteinaceous intracellular inclusions and progressive loss of dopaminergic neurons, which causes motor and non-motor symptoms. Genetic and environmental factors may contribute to the disease etiology. Mitochondrial dysfunction has long been recognized as a hallmark of PD pathogenesis, and several aspects of mitochondrial biology are impaired in PD patients and models. In addition, defects of the autophagy-lysosomal pathway have extensively been observed in cell and animal models as well as PD patients' brains, where constitutive autophagy is indispensable for adaptation to stress and energy deficiency. Genetic and molecular studies have shown that the functions of mitochondria and lysosomal compartments are tightly linked and influence each other. Connections between these organelles are constituted among others by mitophagy, organellar dynamics and cellular signaling cascades, such as calcium (Ca2+) and mTOR (mammalian target of rapamycin) signaling and the activation of transcription factors. Members of the Microphthalmia-associated transcription factor family (MiT), including MITF, TFE3 and TFEB, play a central role in regulating cellular homeostasis in response to metabolic pressure and are considered master regulators of lysosomal biogenesis. As such, they are part of the interconnection between mitochondria and lysosome functions and therefore represent attractive targets for therapeutic approaches against NDD, including PD. The activation of MiT transcription factors through genetic and pharmacological approaches have shown encouraging results at ameliorating PD-related phenotypes in in vitro and in vivo models. In this review, we summarize the relationship between mitochondrial and autophagy-lysosomal functions in the context of PD etiology and focus on the role of the MiT pathway and its potential as pharmacological target against PD.
    Keywords:  Autophagy-lysosomal pathway; Lysosome; MITF; MiT Transcription factors; Mitochondria; Parkinson’s disease; TFE3; TFEB
    DOI:  https://doi.org/10.1186/s13024-022-00555-7
  6. MedComm (2020). 2022 Sep;3(3): e150
    Autophagy in health and disease: From molecular mechanisms to therapeutic target.
    Guang Lu, Yu Wang, Yin Shi, Zhe Zhang, Canhua Huang, Weifeng He, Chuang Wang, Han-Ming Shen.
      Macroautophagy/autophagy is an evolutionally conserved catabolic process in which cytosolic contents, such as aggregated proteins, dysfunctional organelle, or invading pathogens, are sequestered by the double-membrane structure termed autophagosome and delivered to lysosome for degradation. Over the past two decades, autophagy has been extensively studied, from the molecular mechanisms, biological functions, implications in various human diseases, to development of autophagy-related therapeutics. This review will focus on the latest development of autophagy research, covering molecular mechanisms in control of autophagosome biogenesis and autophagosome-lysosome fusion, and the upstream regulatory pathways including the AMPK and MTORC1 pathways. We will also provide a systematic discussion on the implication of autophagy in various human diseases, including cancer, neurodegenerative disorders (Alzheimer disease, Parkinson disease, Huntington's disease, and Amyotrophic lateral sclerosis), metabolic diseases (obesity and diabetes), viral infection especially SARS-Cov-2 and COVID-19, cardiovascular diseases (cardiac ischemia/reperfusion and cardiomyopathy), and aging. Finally, we will also summarize the development of pharmacological agents that have therapeutic potential for clinical applications via targeting the autophagy pathway. It is believed that decades of hard work on autophagy research is eventually to bring real and tangible benefits for improvement of human health and control of human diseases.
    Keywords:  SARS‐CoV‐2; autophagy; cancer; cardiovascular diseases; metabolic diseases; neurodegenerative diseases
    DOI:  https://doi.org/10.1002/mco2.150
  7. Acta Pharm Sin B. 2022 Apr;12(4): 1688-1706
    Pharmacological modulation of autophagy for Alzheimer's disease therapy: Opportunities and obstacles.
    Zhiqiang Deng, Yu Dong, Xiaoting Zhou, Jia-Hong Lu, Zhenyu Yue.
      Alzheimer's disease (AD) is a prevalent and deleterious neurodegenerative disorder characterized by an irreversible and progressive impairment of cognitive abilities as well as the formation of amyloid β (Aβ) plaques and neurofibrillary tangles (NFTs) in the brain. By far, the precise mechanisms of AD are not fully understood and no interventions are available to effectively slow down progression of the disease. Autophagy is a conserved degradation pathway that is crucial to maintain cellular homeostasis by targeting damaged organelles, pathogens, and disease-prone protein aggregates to lysosome for degradation. Emerging evidence suggests dysfunctional autophagy clearance pathway as a potential cellular mechanism underlying the pathogenesis of AD in affected neurons. Here we summarize the current evidence for autophagy dysfunction in the pathophysiology of AD and discuss the role of autophagy in the regulation of AD-related protein degradation and neuroinflammation in neurons and glial cells. Finally, we review the autophagy modulators reported in the treatment of AD models and discuss the obstacles and opportunities for potential clinical application of the novel autophagy activators for AD therapy.
    Keywords:  Alzheimer's disease; Autophagy; Autophagy modulators; Genetic modulation; LC3-associated phagocytosis; Microglial autophagy; Neuroinflammation; Neuronal autophagy
    DOI:  https://doi.org/10.1016/j.apsb.2021.12.009
  8. Autophagy. 2022 Jul 22.
    Atg23 is a vesicle-tethering protein.
    Kelsie A Leary, Wayne D Hawkins, Devika Andhare, Hana Popelka, Daniel J Klionsky, Michael J Ragusa.
      Small 30-nm vesicles containing the integral membrane protein Atg9 provide the initial membrane source for autophagy in yeast. Atg23, is an Atg9 binding protein that is required for Atg9 vesicle trafficking but whose exact function is unknown. In our recent paper, we explored the function of Atg23 using an approach combining cellular biology and biochemistry on purified protein. We determined that Atg23 is an elongated dimer spanning 320 Å in length. We also demonstrated that Atg23 is a membrane-binding and -tethering protein. Furthermore, we identified a series of amino acids residing in a putative coiled- coil region that when mutated prevent Atg23 dimer formation resulting in a stable Atg23 monomer. Last, we demonstrated that when monomeric Atg23 is expressed in yeast lacking Atg23, this leads to a loss of Atg23 puncta, a reduction in Atg9 puncta, a reduction in non-selective autophagy and a complete block in the cytoplasm-to-vacuole targeting (Cvt) pathway.
    Keywords:  Autophagy; cytoplasm-to-vacuole targeting; membrane binding protein; vacuole; yeast
    DOI:  https://doi.org/10.1080/15548627.2022.2105107
  9. Rejuvenation Res. 2022 Jul 19.
    Cellular and mitochondrial quality control mechanisms in maintaining homeostasis in ageing.
    Urvashi Langeh, Vishal Kumar, Anoop Kumar, Pradeep Kumar, Charan Singh, Arti Singh.
      Aging is a natural process in all living organisms defined as destruction of cell function as a result of long-term accumulation of damages. Autophagy is a cellular house safeguard pathway which responsible for degrading damaged cellular organelles. Moreover, it maintains cellular homeostasis, control lifetime, and longevity. Damaged mitochondrial accumulation is a characteristic of aging which associated with neurodegeneration. Mitochondria functions as a principal energy source via supplying ATP through oxidative phosphorylation which serves as fuel for neuronal function. Mitophagy and mitochondrial specific autophagy plays an important role in maintenance of neuronal health via the removal of dysfunctional and aged mitochondria. The mitochondrial QC system involves different strategies for protecting against mitochondrial dysfunction and maintaining healthy mitochondria in cells. Mitochondrial function protection could be a strategy for the promotion of neuroprotection. Mitophagy, could be an effective target for drug discovery. Therefore, further detailed studies for mechanism of mitophagy will advance our mitochondrial phenotype knowledge and understanding to disease pathogenesis. This review mainly focuses on ageing mediated mechanism of autophagy and mitophagy for maintaining the cellular homeostasis and longevity.
    DOI:  https://doi.org/10.1089/rej.2022.0027
  10. Nat Commun. 2022 Jul 21. 13(1): 4220
    Targeting retinoic acid receptor alpha-corepressor interaction activates chaperone-mediated autophagy and protects against retinal degeneration.
    Raquel Gomez-Sintes, Qisheng Xin, Juan Ignacio Jimenez-Loygorri, Mericka McCabe, Antonio Diaz, Thomas P Garner, Xiomaris M Cotto-Rios, Yang Wu, Shuxian Dong, Cara A Reynolds, Bindi Patel, Pedro de la Villa, Fernando Macian, Patricia Boya, Evripidis Gavathiotis, Ana Maria Cuervo.
      Chaperone-mediated autophagy activity, essential in the cellular defense against proteotoxicity, declines with age, and preventing this decline in experimental genetic models has proven beneficial. Here, we have identified the mechanism of action of selective chaperone-mediated autophagy activators previously developed by our group and have leveraged that information to generate orally bioavailable chaperone-mediated autophagy activators with favorable brain exposure. Chaperone-mediated autophagy activating molecules stabilize the interaction between retinoic acid receptor alpha - a known endogenous inhibitor of chaperone-mediated autophagy - and its co-repressor, nuclear receptor corepressor 1, resulting in changes of a discrete subset of the retinoic acid receptor alpha transcriptional program that leads to selective chaperone-mediated autophagy activation. Chaperone-mediated autophagy activators molecules activate this pathway in vivo and ameliorate retinal degeneration in a retinitis pigmentosa mouse model. Our findings reveal a mechanism for pharmacological targeting of chaperone-mediated autophagy activation and suggest a therapeutic strategy for retinal degeneration.
    DOI:  https://doi.org/10.1038/s41467-022-31869-1
  11. Front Aging Neurosci. 2022 ;14 878958
    Induction of Autophagy Promotes Clearance of RHOP23H Aggregates and Protects From Retinal Degeneration.
    Daniela Intartaglia, Giuliana Giamundo, Federica Naso, Edoardo Nusco, Simona Di Giulio, Francesco Giuseppe Salierno, Elena Polishchuk, Ivan Conte.
      Autophagy is a critical metabolic process that acts as a major self-digestion and recycling pathway contributing to maintain cellular homeostasis. An emerging field of research supports the therapeutic modulation of autophagy for treating human neurodegenerative disorders, in which toxic aggregates are accumulated in neurons. Our previous study identified Ezrin protein as an inhibitor of autophagy and lysosomal functions in the retina; thus, in turn, identifying it as a potential pharmacological target for increasing retinal cell clearance to treat inherited retinal dystrophies in which misfolded proteins have accumulated. This study aimed to verify the therapeutic inhibition of Ezrin to induce clearance of toxic aggregates in a mouse model for a dominant form of retinitis pigmentosa (i.e., RHOP23H/+). We found that daily inhibition of Ezrin significantly decreased the accumulation of misfolded RHOP23H aggregates. Remarkably, induction of autophagy, by a drug-mediated pulsatile inhibition of Ezrin, promoted the lysosomal clearance of disease-linked RHOP23H aggregates. This was accompanied with a reduction of endoplasmic reticulum (ER)-stress, robust decrease of photoreceptors' cell death, amelioration in both retinal morphology and function culminating in a better preservation of vision. Our study opens new perspectives for a pulsatile pharmacological induction of autophagy as a mutation-independent therapy paving the way toward a more effective therapeutic strategy to treat these devastating retinal disorders due to an accumulation of intracellular toxic aggregates.
    Keywords:  ER-stress; Ezrin; RHOP23H/+; autophagy; retinal degeneration
    DOI:  https://doi.org/10.3389/fnagi.2022.878958
  12. PLoS Pathog. 2022 Jul 20. 18(7): e1010736
    TBK1 is part of a galectin 8 dependent membrane damage recognition complex and drives autophagy upon Adenovirus endosomal escape.
    Noémie Pied, Coralie F Daussy, Zoé Denis, Jessica Ragues, Muriel Faure, Richard Iggo, Mario P Tschan, Benoit Roger, Fabienne Rayne, Harald Wodrich.
      Intracellular pathogens cause membrane distortion and damage as they enter host cells. Cells perceive these membrane alterations as danger signals and respond by activating autophagy. This response has primarily been studied during bacterial invasion, and only rarely in viral infections. Here, we investigate the cellular response to membrane damage during adenoviral entry. Adenoviruses and their vector derivatives, that are an important vaccine platform against SARS-CoV-2, enter the host cell by endocytosis followed by lysis of the endosomal membrane. We previously showed that cells mount a locally confined autophagy response at the site of endosomal membrane lysis. Here we describe the mechanism of autophagy induction: endosomal membrane damage activates the kinase TBK1 that accumulates in its phosphorylated form at the penetration site. Activation and recruitment of TBK1 require detection of membrane damage by galectin 8 but occur independently of classical autophagy receptors or functional autophagy. Instead, TBK1 itself promotes subsequent autophagy that adenoviruses need to take control of. Depletion of TBK1 reduces LC3 lipidation during adenovirus infection and restores the infectivity of an adenovirus mutant that is restricted by autophagy. By comparing adenovirus-induced membrane damage to sterile lysosomal damage, we implicate TBK1 in the response to a broader range of types of membrane damage. Our study thus highlights an important role for TBK1 in the cellular response to adenoviral endosome penetration and places TBK1 early in the pathway leading to autophagy in response to membrane damage.
    DOI:  https://doi.org/10.1371/journal.ppat.1010736
  13. ACS Omega. 2022 Jul 12. 7(27): 23479-23486
    Development of an RHEB-Targeting Peptide To Inhibit mTORC1 Kinase Activity.
    Raef Shams, Yoshihiro Ito, Hideyuki Miyatake.
      In cancer, the mechanistic/mammalian target of rapamycin complex-1 (mTORC1) is hyperactivated to promote survival under adverse conditions. The kinase activity of mTORC1 is activated by small-GTPase RHEB-GTP. Therefore, a new modality to inhibit mTORC1 activity has emerged, through intercepting RHEB. However, due to the relatively large contact area involved in the interaction between RHEB and mTORC1, facilitating this inhibition through small molecules has been challenging. Here, we report the development of a peptide that can inhibit the RHEB-mTORC1 interaction. The peptide, P1_WT, was designed based on the α-helix (aa 101-115) of the N-heat domain of mTOR to interact with switch II of RHEB. P1_WT bound to RHEB (K D = 0.14 μM) and inhibited RHEB-mTORN-heat interaction (IC50 = 0.33 μM) in vitro. Consequently, P1_WT inhibited mTORC1 activity at a sub-micromolar level (IC50 ∼ 0.3 μM). P1_WT was predicted to be cell-permeable due to the rich content of arginine (23%), enhancing the intracellular translocation. These results show that P1_WT is a potential compound to further develop inhibitors for mTORC1 by intercepting RHEB from mTORC1.
    DOI:  https://doi.org/10.1021/acsomega.2c01865
  14. Front Microbiol. 2022 ;13 889693
    Autophagy Activation Induces p62-Dependent Autophagic Degradation of Dengue Virus Capsid Protein During Infection.
    Yaoxing Wu, Tao Zhou, Jiajia Hu, Yishan Liu, Shouheng Jin, Jianfeng Wu, Xiangdong Guan, Jun Cui.
      In the past decade, dengue virus infection is one of the most prevalent and rapidly spreading arthropod-borne diseases worldwide with about 400 million infections every year. Although it has been reported that the dengue virus could take advantage of autophagy to promote its propagation, the association between selective autophagy and the dengue virus remains largely unclear. Here, we demonstrated that dengue virus capsid protein, the key viral protein for virus assembly, maturation, and replication, underwent autophagic degradation after autophagy activation. Autophagy cargo receptor p62 delivered ubiquitinated capsid protein to autophagosomes for degradation, which could be enhanced by Torin 1 treatments. Further study revealed that the association between p62 and viral capsid protein was dependent on the ubiquitin-binding domain of p62, and the poly-ubiquitin conjugated at lysine 76 of capsid protein served as a recognition signal for autophagy. Consistently, p62 deficiency in Huh7 cells led to the enhancement of dengue virus replication. Our study revealed that p62 targeted dengue virus capsid protein for autophagic degradation in a ubiquitin-dependent manner, which might uncover the potential roles of p62 in restricting dengue virus replication.
    Keywords:  autophagic degradation; capsid protein (C protein); cargo receptors; dengue virus; p62 (SQSTM1); ubiquitination
    DOI:  https://doi.org/10.3389/fmicb.2022.889693
  15. Autophagy. 2022 Jul 22. 1-18
    Stimulation of synaptic activity promotes TFEB-mediated clearance of pathological MAPT/Tau in cellular and mouse models of tauopathies.
    Yvette Akwa, Chiara Di Malta, Fátima Zallo, Elise Gondard, Adele Lunati, Lara Z Diaz-de-Grenu, Angela Zampelli, Anne Boiret, Sara Santamaria, Maialen Martinez-Preciado, Katia Cortese, Jeffrey H Kordower, Carlos Matute, Andres M Lozano, Estibaliz Capetillo-Zarate, Thomas Vaccari, Carmine Settembre, Etienne E Baulieu, Davide Tampellini.
      Synapses represent an important target of Alzheimer disease (AD), and alterations of their excitability are among the earliest changes associated with AD development. Synaptic activation has been shown to be protective in models of AD, and deep brain stimulation (DBS), a surgical strategy that modulates neuronal activity to treat neurological and psychiatric disorders, produced positive effects in AD patients. However, the molecular mechanisms underlying the protective role(s) of brain stimulation are still elusive. We have previously demonstrated that induction of synaptic activity exerts protection in mouse models of AD and frontotemporal dementia (FTD) by enhancing the macroautophagy/autophagy flux and lysosomal degradation of pathological MAPT/Tau. We now provide evidence that TFEB (transcription factor EB), a master regulator of lysosomal biogenesis and autophagy, is a key mediator of this cellular response. In cultured primary neurons from FTD-transgenic mice, synaptic stimulation inhibits MTORC1 signaling, thus promoting nuclear translocation of TFEB, which, in turn, induces clearance of MAPT/Tau oligomers. Conversely, synaptic activation fails to promote clearance of toxic MAPT/Tau in neurons expressing constitutively active RRAG GTPases, which sequester TFEB in the cytosol, or upon TFEB depletion. Activation of TFEB is also confirmed in vivo in DBS-stimulated AD mice. We also demonstrate that DBS reduces pathological MAPT/Tau and promotes neuroprotection in Parkinson disease patients with tauopathy. Altogether our findings indicate that stimulation of synaptic activity promotes TFEB-mediated clearance of pathological MAPT/Tau. This mechanism, underlying the protective effect of DBS, provides encouraging support for the use of synaptic stimulation as a therapeutic treatment against tauopathies.
    Keywords:  Alzheimer; autophagy; deep brain stimulation; lysosome; neuron; synapse; tau
    DOI:  https://doi.org/10.1080/15548627.2022.2095791
  16. eNeuro. 2022 Jul 18. pii: ENEURO.0480-21.2022. [Epub ahead of print]
    Confirmation of Decreased Rates of Cerebral Protein Synthesis In Vivo in a Mouse Model of Tuberous Sclerosis Complex.
    Rachel Michelle Saré, Anita Torossian, Inna Loutaev, Carolyn Beebe Smith.
      Tuberous sclerosis complex (TSC) is an autosomal dominant disorder that results in intellectual disability and, in ∼50% of patients, autism spectrum disorder. The protein products that are altered in TSC (TSC1 and TSC2) form a complex to inhibit the mammalian target of rapamycin [mTOR; mTOR complex 1 (mTORC1)] pathway. This pathway has been shown to affect the process of mRNA translation through its action on ribosomal protein S6 and 4-elongation binding protein 1. It is thought that mutations in the TSC proteins lead to upregulation of the mTORC1 pathway and consequently an increase in protein synthesis. Unexpectedly, our previous study of a mouse model of TSC (Tsc2Djk +/) demonstrated decreased in vivo rates of protein synthesis throughout the brain. In the present study, we confirm those results in another Tsc2+/- mouse model, one with a different mutation locus and on a mixed background (Tsc2Mjg +/-). We also examine mTORC1 signaling and possible effects of prior isoflurane anesthesia. Because measurements of protein synthesis rates in vivo require surgical preparation of the animal and anesthesia, we examine mTORC1 signaling pathways both under baseline conditions and following recovery from anesthesia. Our results demonstrate regionally selective effects of prior anesthesia. Overall, our results in both in vivo models suggest differences to the central hypothesis regarding TSC and show the importance of studying protein synthesis in vivo Significance StatementProtein synthesis is an important process for brain function. In the disorder, tuberous sclerosis complex (TSC), the inhibition of the mammalian target of rapamycin (mTOR) pathway is reduced and this is thought to lead to excessive protein synthesis. Most studies of protein synthesis in models of TSC have been conducted in vitro We report here confirmation of our previous in vivo study showing decreased brain protein synthesis rates in a second mouse model of TSC, results counter to the central hypothesis regarding TSC. We also explore the possible influence of prior isoflurane exposure on signaling pathways involved in regulation of protein synthesis. This study highlights a novel aspect of TSC and the importance of studying cellular processes in vivo.
    Keywords:  mTOR; protein synthesis; tuberous sclerosis
    DOI:  https://doi.org/10.1523/ENEURO.0480-21.2022
  17. Angew Chem Int Ed Engl. 2022 Jul 22.
    An ER-targeting DNA Nanodevice for Autophagy-dependent Degradation of Proteins in Membrane-bound Organelles.
    Caixia Liu, Bin Wang, Weiping Zhu, Yufang Xu, Yangyang Yang, Xuhong Qian.
      Targeted protein degradation via proteasomal and lysosomal pathways has become a promising therapeutic approach, and proteins in cytoplasm or on the cell membrane can be easily contacted and have become the major targets. However, degradation of disease-related proteins that exist in membrane-bound organelles (MBO) such as the endoplasmic reticulum (ER) remains unsolved due to the membrane limits. Here we describe a DNA nanodevice that shows ER targeting capacity and undergoes new intracellular degradation via the autophagy-dependent pathway. Then the DNA nanostructure functionalized with specific ligands are used to selectively catch ER-localized proteins and then transport them to the lysosome for degradation. Through this technique, the degradation of both exogenous ER-resident protein (ER-eGFP) and endogenous overexpressed molecular chaperone (glucose-regulated protein 78) in cancer cells has been successfully executed with high efficiency. This strategy expands the range of currently proposed protein degradation platforms and may contribute to the development of targeted cancer therapeutics.
    Keywords:  Autophagy; DNA nanostructure; Organelle targeting; Targeted protein degradation
    DOI:  https://doi.org/10.1002/anie.202205509
  18. Nat Metab. 2022 Jul 21.
    TFEB induces mitochondrial itaconate synthesis to suppress bacterial growth in macrophages.
    Ev-Marie Schuster, Maximilian W Epple, Katharina M Glaser, Michael Mihlan, Kerstin Lucht, Julia A Zimmermann, Anna Bremser, Aikaterini Polyzou, Nadine Obier, Nina Cabezas-Wallscheid, Eirini Trompouki, Andrea Ballabio, Jörg Vogel, Joerg M Buescher, Alexander J Westermann, Angelika S Rambold.
      Successful elimination of bacteria in phagocytes occurs in the phago-lysosomal system, but also depends on mitochondrial pathways. Yet, how these two organelle systems communicate is largely unknown. Here we identify the lysosomal biogenesis factor transcription factor EB (TFEB) as regulator for phago-lysosome-mitochondria crosstalk in macrophages. By combining cellular imaging and metabolic profiling, we find that TFEB activation, in response to bacterial stimuli, promotes the transcription of aconitate decarboxylase (Acod1, Irg1) and synthesis of its product itaconate, a mitochondrial metabolite with antimicrobial activity. Activation of the TFEB-Irg1-itaconate signalling axis reduces the survival of the intravacuolar pathogen Salmonella enterica serovar Typhimurium. TFEB-driven itaconate is subsequently transferred via the Irg1-Rab32-BLOC3 system into the Salmonella-containing vacuole, thereby exposing the pathogen to elevated itaconate levels. By activating itaconate production, TFEB selectively restricts proliferating Salmonella, a bacterial subpopulation that normally escapes macrophage control, which contrasts TFEB's role in autophagy-mediated pathogen degradation. Together, our data define a TFEB-driven metabolic pathway between phago-lysosomes and mitochondria that restrains Salmonella Typhimurium burden in macrophages in vitro and in vivo.
    DOI:  https://doi.org/10.1038/s42255-022-00605-w
  19. 3 Biotech. 2022 Aug;12(8): 174
    Frustration analysis of TBK1 missense mutations reported in ALS/FTD and cancer patients.
    Fatima Khatoon, Vijay Kumar, Farah Anjum, Alaa Shafie, Mohd Adnan, Md Imtaiyaz Hassan.
      Tank-binding kinase 1 (TBK1) is a multifunctional kinase having essential roles in cellular processes, autophagy/mitophagy, and selective clearance of damaged proteins. More than 90 mutations in the TBK1 gene are linked with multiple cancer types, amyotrophic lateral sclerosis (ALS), and frontotemporal dementia (FTD). Some of these missense mutations disrupt the abilities of TBK1 to dimerize, associate with the mitophagy receptor optineurin (OPTN), autoactivate, or catalyze phosphorylation. Some mutations may cause severe dysregulation of the pathway, while others induce a limited disruption. Here, we have studied those mutations reported in cancer, ALS and FTD, and subsequently investigated the effect of missense mutations on the structure and function of TBK1 for localized residual frustration change. Out of 33 ALS/FTD causing mutations and 28 oncogenic mutations, 10 mutations and 12 oncogenic mutations showed significant change in the residual frustration. The local frustration plays an important role in the conformation of protein structure in active and inactive kinases. Our analysis reports the change in residual frustration state, conformational change and effect on active and inactive TBK1 function due to ALS/FTD causing and oncogenic missense mutations.
    Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03240-0.
    Keywords:  Amyotrophic lateral sclerosis; Autophagy; Cancer; Comparative genomics; Fronto-temporal dementia; Local frustration; Tank-binding kinase 1
    DOI:  https://doi.org/10.1007/s13205-022-03240-0
  20. Redox Biol. 2022 Jul 14. pii: S2213-2317(22)00179-3. [Epub ahead of print]55 102407
    Integrated regulation of stress responses, autophagy and survival by altered intracellular iron stores.
    Yunyang Wang, Mo Wang, Yunshan Liu, Hui Tao, Somesh Banerjee, Shanthi Srinivasan, Elizabeta Nemeth, Mark J Czaja, Peijian He.
      Iron is a mineral essential for blood production and a variety of critical cellular functions. Altered iron metabolism has been increasingly observed in many diseases and disorders, but a comprehensive and mechanistic understanding of the cellular impact of impaired iron metabolism is still lacking. We examined the effects of iron overload or iron deficiency on cellular stress responses and autophagy which collectively regulate cell homeostasis and survival. Acute iron loading led to increased mitochondrial ROS (mtROS) production and damage, lipid peroxidation, impaired autophagic flux, and ferroptosis. Iron-induced mtROS overproduction is the mechanism of increased lipid peroxidation, impaired autophagy, and the induction of ferroptosis. Iron excess-induced ferroptosis was cell-type dependent and regulated by activating transcription factor 4 (ATF4). Upregulation of ATF4 mitigated iron-induced autophagic dysfunction and ferroptosis, whereas silencing of ATF4 expression impaired autophagy and resulted in increased mtROS production and ferroptosis. Employing autophagy-deficient hepatocytes and different autophagy inhibitors, we further showed that autophagic impairment sensitized cells to iron-induced ferroptosis. In contrast, iron deficiency activated the endoplasmic reticulum (ER) stress response, decreased autophagy, and induced apoptosis. Decreased autophagy associated with iron deficiency was due to ER stress, as reduction of ER stress by 4-phenylbutyric acid (4-PBA) improved autophagic flux. The mechanism of decreased autophagy in iron deficiency is a disruption in lysosomal biogenesis due to impaired posttranslational maturation of lysosomal membrane proteins. In conclusion, iron excess and iron deficiency cause different forms of cell stress and death in part through the common mechanism of impaired autophagic function.
    Keywords:  ATF4; ER stress; Ferroptosis; Lipid peroxidation; Mitochondria
    DOI:  https://doi.org/10.1016/j.redox.2022.102407
  21. Acta Pharm Sin B. 2022 Apr;12(4): 1707-1722
    Celastrol enhances transcription factor EB (TFEB)-mediated autophagy and mitigates Tau pathology: Implications for Alzheimer's disease therapy.
    Chuanbin Yang, Chengfu Su, Ashok Iyaswamy, Senthil Kumar Krishnamoorthi, Zhou Zhu, Sichang Yang, Benjamin Chunkit Tong, Jia Liu, Sravan G Sreenivasmurthy, Xinjie Guan, Yuxuan Kan, Aston Jiaxi Wu, Alexis Shiying Huang, Jieqiong Tan, Kingho Cheung, Juxian Song, Min Li.
      Alzheimer's disease (AD), characterized by the accumulation of protein aggregates including phosphorylated Tau aggregates, is the most common neurodegenerative disorder with limited therapeutic agents. Autophagy plays a critical role in the degradation of phosphorylated Tau aggregates, and transcription factor EB (TFEB) is a master regulator of autophagy and lysosomal biogenesis. Thus, small-molecule autophagy enhancers targeting TFEB hold promise for AD therapy. Here, we found that celastrol, an active ingredient isolated from the root extracts of Tripterygium wilfordii (Lei Gong Teng in Chinese) enhanced TFEB-mediated autophagy and lysosomal biogenesis in vitro and in mouse brains. Importantly, celastrol reduced phosphorylated Tau aggregates and attenuated memory dysfunction and cognitive deficits in P301S Tau and 3xTg mice, two commonly used AD animal models. Mechanistical studies suggest that TFEB-mediated autophagy-lysosomal pathway is responsible for phosphorylated Tau degradation in response to celastrol. Overall, our findings indicate that Celastrol is a novel TFEB activator that promotes the degradation of phosphorylated Tau aggregates and improves memory in AD animal models. Therefore, Celastrol shows potential as a novel agent for the treatment and/or prevention of AD and other tauopathies.
    Keywords:  Alzheimer's disease (AD); Autophagy; Celastrol; Lysosome biogenesis; TFEB; Tau; Therapeutic target; mTOR
    DOI:  https://doi.org/10.1016/j.apsb.2022.01.017
  22. Oxid Med Cell Longev. 2022 ;2022 7925686
    MANF Inhibits α-Synuclein Accumulation through Activation of Autophagic Pathways.
    Jing-Xing Zhang, Wei-Fang Tong, Ming Jiang, Kai-Ge Zhou, Xue-Rui Xiang, Yi-Jing He, Zhuo-Yu Zhang, Qiang Guan, Ling-Jing Jin.
      Progressive accumulation of misfolded SNCA/α-synuclein is key to the pathology of Parkinson's disease (PD). Drugs aiming at degrading SNCA may be an efficient therapeutic strategy for PD. Our previous study showed that mesencephalic astrocyte-derived neurotrophic factor (MANF) facilitated the removal of misfolded SNCA and rescued dopaminergic (DA) neurons, but the underlying mechanisms remain unknown. In this study, we showed that AAV8-MANF relieved Parkinsonian behavior in rotenone-induced PD model and reduced SNCA accumulation in the substantia nigra. By establishing wildtype (WT) SNCA overexpression cellular model, we found that chaperone-mediated-autophagy (CMA) and macroautophagy were both participated in MANF-mediated degradation of SNCAWT. Nuclear factor erythroid 2-related factor (Nrf2) was activated to stimulating macroautophagy activity when CMA pathway was impaired. Using A53T mutant SNCA overexpression cellular model to mimic CMA dysfunction situation, we concluded that macroautophagy rather than CMA was responsible to the degradation of SNCAA53T, and this degradation was mediated by Nrf2 activation. Hence, our findings suggested that MANF has potential therapeutic value for PD. Nrf2 and its role in MANF-mediated degradation may provide new sights that target degradation pathways to counteract SNCA pathology in PD.
    DOI:  https://doi.org/10.1155/2022/7925686
  23. Nat Commun. 2022 Jul 16. 13(1): 4146
    Compounds activating VCP D1 ATPase enhance both autophagic and proteasomal neurotoxic protein clearance.
    Lidia Wrobel, Sandra M Hill, Alvin Djajadikerta, Marian Fernandez-Estevez, Cansu Karabiyik, Avraham Ashkenazi, Victoria J Barratt, Eleanna Stamatakou, Anders Gunnarsson, Timothy Rasmusson, Eric W Miele, Nigel Beaton, Roland Bruderer, Yuehan Feng, Lukas Reiter, M Paola Castaldi, Rebecca Jarvis, Keith Tan, Roland W Bürli, David C Rubinsztein.
      Enhancing the removal of aggregate-prone toxic proteins is a rational therapeutic strategy for a number of neurodegenerative diseases, especially Huntington's disease and various spinocerebellar ataxias. Ideally, such approaches should preferentially clear the mutant/misfolded species, while having minimal impact on the stability of wild-type/normally-folded proteins. Furthermore, activation of both ubiquitin-proteasome and autophagy-lysosome routes may be advantageous, as this would allow effective clearance of both monomeric and oligomeric species, the latter which are inaccessible to the proteasome. Here we find that compounds that activate the D1 ATPase activity of VCP/p97 fulfill these requirements. Such effects are seen with small molecule VCP activators like SMER28, which activate autophagosome biogenesis by enhancing interactions of PI3K complex components to increase PI(3)P production, and also accelerate VCP-dependent proteasomal clearance of such substrates. Thus, this mode of VCP activation may be a very attractive target for many neurodegenerative diseases.
    DOI:  https://doi.org/10.1038/s41467-022-31905-0
  24. Cell Death Dis. 2022 Jul 18. 13(7): 622
    Transcription factor EB-mediated mesenchymal stem cell therapy induces autophagy and alleviates spinocerebellar ataxia type 3 defects in neuronal cells model.
    Xiaobo Han, Jean de Dieu Habimana, Amy L Li, Rongqi Huang, Omar Mukama, Weiyue Deng, Ling Wang, Yuying Zhang, Wei Wang, Sihao Deng, Kexin Peng, Bing Ni, Shusheng Zhang, Jufang Huang, Xiao-Xin Yan, Zhiyuan Li.
      Defects in ataxin-3 proteins and CAG repeat expansions in its coding gene ATXN3 cause Spinocerebellar Ataxia Type 3 (SCA3) or Machado-Joseph disease (MJD) polyglutamine neurodegenerative disease. The mutant proteins aggregate as inclusion bodies in cells and compete with wild-type ataxin-3, which leads to neuronal dysfunction or death and impairs Beclin1-mediated autophagy. It has been reported that Mesenchymal stem cells (MSCs) can reliably treat several neurodegenerative diseases. Herein, we used a Transcription Factor EB (TFEB) nuclear translocation-mediated MSCs co-culture approach to reconstitute autophagy and lysosomal biogenesis, and reduce SCA3-like behaviors in induced pluripotent stem cells (iPSCs)-derived neuron cells models. Our iPSCs model showed enhanced expression of autophagy proteins, attenuated the expression and toxic effects of mutant ataxin-3 on neurons, and alleviated the effects of ataxin-3 on autophagy. Therefore, MSCs are associated with autophagy-inducing therapy and compared to animal models, our MSCs co-culture could be used as a novel and potential therapeutic approach to study SCA3 disease and other neurodegenerative diseases.
    DOI:  https://doi.org/10.1038/s41419-022-05085-0
  25. Methods Mol Biol. 2022 ;2535 211-220
    Analysis of ER-Phagy in Cancer Drug Resistance.
    Sandhya Chipurupalli, Vincenzo Desiderio, Nirmal Robinson.
      The ability of the cancer cells to survive hostile environment depends on their cellular stress response mechanisms. These mechanisms also help them to develop resistance to chemotherapies. Autophagy and more specifically organelle specific autophagy is one such adaptive mechanism that promotes drug resistance in cancer cells. Endoplasmic reticulum-specific autophagy or ER-phagy has been more recently described to overcome ER-stress through the degradation of damaged ER. ER-resident proteins such as FAM134B act as ER-phagy receptors to specifically target damaged ER for degradation through autophagy. Moreover, we had recently deciphered that ER-phagy facilitates cancer cell survival during hypoxic stress and we predict that this process could play a critical role in the development of drug resistance in cancer cells. Therefore, here, we provide a lay description of how ER-phagy could be investigated biochemically by Western blot analysis and silencing ER-phagy receptor genes using small interfering RNAs (siRNA).
    Keywords:  Autophagy; Cancer; Chemotherapy; Drug resistance; ER-phagy; FAM134B; Hypoxia; LC3
    DOI:  https://doi.org/10.1007/978-1-0716-2513-2_16
  26. Autophagy. 2022 Jul 20. 1-16
    Lack of COL6/collagen VI causes megakaryocyte dysfunction by impairing autophagy and inducing apoptosis.
    Vittorio Abbonante, Alessandro Malara, Martina Chrisam, Samuele Metti, Paolo Soprano, Claudio Semplicini, Luca Bello, Valeria Bozzi, Monica Battiston, Alessandro Pecci, Elena Pegoraro, Luigi De Marco, Paola Braghetta, Paolo Bonaldo, Alessandra Balduini.
      Endoplasmic reticulum stress is an emerging significant player in the molecular pathology of connective tissue disorders. In response to endoplasmic reticulum stress, cells can upregulate macroautophagy/autophagy, a fundamental cellular homeostatic process used by cells to degrade and recycle proteins or remove damaged organelles. In these scenarios, autophagy activation can support cell survival. Here we demonstrated by in vitro and in vivo approaches that megakaryocytes derived from col6a1-⁄- (collagen, type VI, alpha 1) null mice display increased intracellular retention of COL6 polypeptides, endoplasmic reticulum stress and apoptosis. The unfolded protein response is activated in col6a1-⁄- megakaryocytes, as evidenced by the upregulation of molecular chaperones, by the increased splicing of Xbp1 mRNA and by the higher level of the pro-apoptotic regulator DDIT3/CHOP. Despite the endoplasmic reticulum stress, basal autophagy is impaired in col6a1-⁄- megakaryocytes, which show lower BECN1 levels and reduced autophagosome maturation. Starvation and rapamycin treatment rescue the autophagic flux in col6a1-⁄- megakaryocytes, leading to a decrease in intracellular COL6 polypeptide retention, endoplasmic reticulum stress and apoptosis. Furthermore, megakaryocytes cultured from peripheral blood hematopoietic progenitors of patients affected by Bethlem myopathy and Ullrich congenital muscular dystrophy, two COL6-related disorders, displayed increased apoptosis, endoplasmic reticulum stress and impaired autophagy. These data demonstrate that genetic disorders of collagens, endoplasmic reticulum stress and autophagy regulation in megakaryocytes may be interrelated.Abbreviations: 7-AAD: 7-amino-actinomycin D; ATF: activating transcriptional factor; BAX: BCL2 associated X protein; BCL2: B cell leukemia/lymphoma 2; BCL2L1/Bcl-xL: BCL2-like 1; BM: bone marrow; COL6: collagen, type VI; col6a1-⁄-: mice that are null for Col6a1; DDIT3/CHOP/GADD153: DNA-damage inducible transcript 3; EGFP: enhanced green fluorescent protein; ER: endoplasmic reticulum; reticulophagy: endoplasmic reticulum-selective autophagy; HSPA5/Bip: heat shock protein 5; HSP90B1/GRP94: heat shock protein 90, beta (Grp94), member 1; LAMP2: lysosomal associated membrane protein 2; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; Mk: megakaryocytes; MTOR: mechanistic target of rapamycin kinase; NIMV: noninvasive mechanical ventilation; PI3K: phosphoinositide 3-kinase; PPP1R15A/GADD34: protein phosphatase 1, regulatory subunit 15A; RT-qPCR: reverse transcription-quantitative real-time PCR; ROS: reactive oxygen species; SERPINH1/HSP47: serine (or cysteine) peptidase inhibitor, clade H, member 1; sh-RNA: short hairpin RNA; SOCE: store operated calcium entry; UCMD: Ullrich congenital muscular dystrophy; UPR: unfolded protein response; WIPI2: WD repeat domain, phosphoinositide-interacting 2; WT: wild type; XBP1: X-box binding protein 1.
    Keywords:  Apoptosis; autophagy; collagen VI; endoplasmic reticulum stress; megakaryocytes; rapamycin; unfolded protein response
    DOI:  https://doi.org/10.1080/15548627.2022.2100105
  27. Nutrition. 2022 Apr 11. pii: S0899-9007(22)00098-3. [Epub ahead of print]103-104 111686
    Nutritional strategies for autophagy activation and health consequences of autophagy impairment.
    Aleksandra M Kocot, Barbara Wróblewska.
       OBJECTIVES: Currently, the plague of chronic diseases, such as overweight, obesity, diabetes, and cardiovascular diseases, is associated with chronic inflammation as an effect of homeostasis disbalance. One of the processes involved in homeostasis maintenance is autophagy, which is also referred to as self-eating or cellular recycling. Due to the correlation between the epidemic scale of chronic diseases and autophagy impairment, strategies for autophagy activation are urgently needed.
    METHODS: In this review, we comprehensively summarized the current data on autophagy types, dysfunction, associated diseases, and ways of activation available in scientific databases and published up until 2022.
    RESULTS: Our review indicates that impaired autophagy is associated with inflammatory bowel diseases, cancer, overweight, obesity, type I diabetes, diseases of the cardiovascular system, neurodegenerative diseases, depression, and anxiety. We highlight nutritional behavior as one of the factors behind autophagy induction and homeostasis restoration.
    CONCLUSIONS: Autophagy is involved in different dysfunctions and diseases; thus, activation strategies are urgently needed. A high potential in the prevention and therapies of chronic diseases by means of autophagy induction can be expected from nutritional behaviors. To date, most studies were carried out in vitro or in a murine model. Thus, further, well-designed, clinical trials are needed to provide the missing understanding of the nutritional potential to regulate specific signaling pathways that keep autophagy running smoothly.
    Keywords:  Autophagy activation, Autophagy-related diseases; Human well-being; Nutritional strategies; Nutritional support
    DOI:  https://doi.org/10.1016/j.nut.2022.111686
  28. Front Cell Infect Microbiol. 2022 ;12 892610
    Interaction Between Autophagy and Porphyromonas gingivalis-Induced Inflammation.
    Sen Kang, Anna Dai, Huiming Wang, Pei-Hui Ding.
      Autophagy is an immune homeostasis process induced by multiple intracellular and extracellular signals. Inflammation is a protective response to harmful stimuli such as pathogen microbial infection and body tissue damage. Porphyromonas gingivalis infection elicits both autophagy and inflammation, and dysregulation of autophagy and inflammation promotes pathology. This review focuses on the interaction between autophagy and inflammation caused by Porphyromonas gingivalis infection, aiming to elaborate on the possible mechanism involved in the interaction.
    Keywords:  Porphyromonas gingivalis; autophagy; inflammation; macroautophagy; xenophagy
    DOI:  https://doi.org/10.3389/fcimb.2022.892610
  29. Cell Rep. 2022 Jul 19. pii: S2211-1247(22)00930-5. [Epub ahead of print]40(3): 111124
    Pathological mitophagy disrupts mitochondrial homeostasis in Leber's hereditary optic neuropathy.
    Alberto Danese, Simone Patergnani, Alessandra Maresca, Camille Peron, Andrea Raimondi, Leonardo Caporali, Saverio Marchi, Chiara La Morgia, Valentina Del Dotto, Claudia Zanna, Angelo Iannielli, Alice Segnali, Ivano Di Meo, Andrea Cavaliere, Magdalena Lebiedzinska-Arciszewska, Mariusz R Wieckowski, Andrea Martinuzzi, Milton N Moraes-Filho, Solange R Salomao, Adriana Berezovsky, Rubens Belfort, Christopher Buser, Fred N Ross-Cisneros, Alfredo A Sadun, Carlo Tacchetti, Vania Broccoli, Carlotta Giorgi, Valeria Tiranti, Valerio Carelli, Paolo Pinton.
      Leber's hereditary optic neuropathy (LHON), a disease associated with a mitochondrial DNA mutation, is characterized by blindness due to degeneration of retinal ganglion cells (RGCs) and their axons, which form the optic nerve. We show that a sustained pathological autophagy and compartment-specific mitophagy activity affects LHON patient-derived cells and cybrids, as well as induced pluripotent-stem-cell-derived neurons. This is variably counterbalanced by compensatory mitobiogenesis. The aberrant quality control disrupts mitochondrial homeostasis as reflected by defective bioenergetics and excessive reactive oxygen species production, a stress phenotype that ultimately challenges cell viability by increasing the rate of apoptosis. We counteract this pathological mechanism by using autophagy regulators (clozapine and chloroquine) and redox modulators (idebenone), as well as genetically activating mitochondrial biogenesis (PGC1-α overexpression). This study substantially advances our understanding of LHON pathophysiology, providing an integrated paradigm for pathogenesis of mitochondrial diseases and druggable targets for therapy.
    Keywords:  CP: Neuroscience; LHON; autophagy; cybrids; iPSCs; mitochondria; mitophagy; mtDNA; optic nerve; retinal ganglion cells; therapy
    DOI:  https://doi.org/10.1016/j.celrep.2022.111124
  30. Mov Disord. 2022 Jul 20.
    Rest and Digest-The Basal Role of Autophagy in Neurons and Its Relevance to Parkinson's Disease.
    Neda M Ilieva, Briana R De Miranda.
      
    DOI:  https://doi.org/10.1002/mds.29165
  31. Acta Pharm Sin B. 2022 Jul;12(7): 3085-3102
    Repurposing econazole as a pharmacological autophagy inhibitor to treat pancreatic ductal adenocarcinoma.
    Ningna Weng, Siyuan Qin, Jiayang Liu, Xing Huang, Jingwen Jiang, Li Zhou, Zhe Zhang, Na Xie, Kui Wang, Ping Jin, Maochao Luo, Liyuan Peng, Edouard C Nice, Ajay Goel, Suxia Han, Canhua Huang, Qing Zhu.
      Pancreatic ductal adenocarcinoma (PDAC) is characterized by the highest mortality among carcinomas. The pathogenesis of PDAC requires elevated autophagy, inhibition of which using hydroxychloroquine has shown promise. However, current realization is impeded by its suboptimal use and unpredictable toxicity. Attempts to identify novel autophagy-modulating agents from already approved drugs offer a rapid and accessible approach. Here, using a patient-derived organoid model, we performed a comparative analysis of therapeutic responses among various antimalarial/fungal/parasitic/viral agents, through which econazole (ECON), an antifungal compound, emerged as the top candidate. Further testing in cell-line and xenograft models of PDAC validated this activity, which occurred as a direct consequence of dysfunctional autophagy. More specifically, ECON boosted autophagy initiation but blocked lysosome biogenesis. RNA sequencing analysis revealed that this autophagic induction was largely attributed to the altered expression of activation transcription factor 3 (ATF3). Increased nuclear import of ATF3 and its transcriptional repression of inhibitor of differentiation-1 (ID-1) led to inactivation of the AKT/mammalian target of rapamycin (mTOR) pathway, thus giving rise to autophagosome accumulation in PDAC cells. The magnitude of the increase in autophagosomes was sufficient to elicit ER stress-mediated apoptosis. Furthermore, ECON, as an autophagy inhibitor, exhibited synergistic effects with trametinib on PDAC. This study provides direct preclinical and experimental evidence for the therapeutic efficacy of ECON in PDAC treatment and reveals a mechanism whereby ECON inhibits PDAC growth.
    Keywords:  AKT; ATF3; Autophagy; Econazole; Organoid; PDAC; Therapy; Trametinib
    DOI:  https://doi.org/10.1016/j.apsb.2022.01.018
  32. Hepatology. 2022 Jul 21.
    Hepatocyte β-Catenin Loss is Compensated by Insulin-mTORC1 Activation to Promote Liver Regeneration.
    Shikai Hu, Catherine Cao, Minakshi Poddar, Evan Delgado, Sucha Singh, Anya Singh-Varma, Donna Beer Stolz, Aaron Bell, Satdarshan P Monga.
       BACKGROUND AND AIMS: Liver regeneration (LR) following partial hepatectomy (PH) occurs via activation of various signaling pathways. Disruption of single pathway can be compensated by activation of another pathway to continue LR. The Wnt-β-catenin pathway is activated early during LR and conditional hepatocyte loss of β-catenin delays LR. Here, we study mechanism of LR in the absence of hepatocyte-β-catenin.
    APPROACH & RESULTS: Eight-week-old hepatocyte-specific Ctnnb1 knockout mice (β-cateninΔHC ) were subjected to PH. These animals exhibited decreased hepatocyte proliferation at 40-120h and decreased cumulative 14-day (14d) BrdU labeling of <40%, but all mice survived suggesting compensation. Insulin-mediated mTORC1 activation was uniquely identified in the β-cateninΔHC mice at 72-96h after PH. Deletion of hepatocyte Raptor, a critical mTORC1 partner, in the β-cateninΔHC mice led to progressive hepatic injury and mortality by 30d. PH on early-stage non-morbid RaptorΔHC -β-cateninΔHC mice led to lethality by 12h. RaptorΔHC mice showed progressive hepatic injury, spontaneous LR with β-catenin activation, but died by 40d. PH on early stage non-morbid RaptorΔHC mice was lethal by 48h. Temporal inhibition of insulin receptor and mTORC1 in β-cateninΔHC or controls after PH was achieved by administration of linsitinib at 48h or rapamycin at 60h post-PH, and completely prevented LR leading to lethality by 12-14d.
    CONCLUSIONS: Insulin-mTORC1 activation compensates for β-catenin loss to enable LR after PH. mTORC1 signaling in hepatocytes itself is critical to both homeostasis and LR and is only partially compensated by β-catenin activation. Dual inhibition of β-catenin and mTOR may have notable untoward hepatotoxic side effects.
    DOI:  https://doi.org/10.1002/hep.32680
  33. Proc Natl Acad Sci U S A. 2022 Jul 26. 119(30): e2201927119
    Autophagic membranes participate in hepatitis B virus nucleocapsid assembly, precore and core protein trafficking, and viral release.
    Ja Yeon Kim Chu, Yu-Chen Chuang, Kuen-Nan Tsai, Jessica Pantuso, Yuji Ishida, Takeshi Saito, Jing-Hsiung James Ou.
      Hepatitis B virus (HBV) DNA replication takes place inside the viral core particle and is dependent on autophagy. Here we show that HBV core particles are associated with autophagosomes and phagophores in cells that productively replicate HBV. These autophagic membrane-associated core particles contain almost entirely the hypophosphorylated core protein and are DNA replication competent. As the hyperphosphorylated core protein can be localized to phagophores and the dephosphorylation of the core protein is associated with the packaging of viral pregenomic RNA (pgRNA), these results are in support of the model that phagophores can serve as the sites for the packaging of pgRNA. In contrast, in cells that replicate HBV, the precore protein derivatives, which are related to the core protein, are associated with autophagosomes but not with phagophores via a pathway that is independent of its signal peptide. Interestingly, when the core protein is expressed by itself, it is associated with phagophores but not with autophagosomes. These observations indicate that autophagic membranes are differentially involved in the trafficking of precore and core proteins. HBV induces the fusion of autophagosomes and multivesicular bodies and the silencing of Rab11, a regulator of this fusion, is associated with the reduction of release of mature HBV particles. Our studies thus indicate that autophagic membranes participate in the assembly of HBV nucleocapsids, the trafficking of HBV precore and core proteins, and likely also the egress of HBV particles.
    Keywords:  HBV protein trafficking; autophagosomes; autophagy; hepatitis B virus; phagophores
    DOI:  https://doi.org/10.1073/pnas.2201927119
  34. J Biol Chem. 2022 Jul 13. pii: S0021-9258(22)00702-5. [Epub ahead of print] 102260
    A small molecule Toll-like receptor antagonist rescues α-synuclein fibril pathology.
    Jessica Chedid, Adahir Labrador-Garrido, Siying Zhong, Jianqun Gao, Ye Zhao, Gayathri Perera, Woojin S Kim, Glenda M Halliday, Nicolas Dzamko.
      The propagation and accumulation of pathological α-synuclein protein is thought to underlie the clinical symptoms of the neurodegenerative movement disorder Parkinson's disease (PD). Consequently, there is significant interest in identifying the mechanisms that contribute to α-synuclein pathology, as these may inform therapeutic targets for the treatment of PD. One protein that appears to contribute to α-synuclein pathology is the innate immune pathogen recognition receptor, Toll-like receptor 2 (TLR). TLR2 is expressed on neurons and its activation results in the accumulation of α-synuclein protein; however, the precise mechanism by which TLR2 contributes to α-synuclein pathology is unclear. Herein we demonstrate using human cell models that neuronal TLR2 activation acutely impairs the autophagy lysosomal pathway and markedly potentiates α-synuclein pathology seeded with α-synuclein pre-formed fibrils (PFF). Moreover, α-synuclein pathology could be ameliorated with a novel small molecule TLR2 inhibitor, including in induced pluripotent stem cell-derived neurons from a patient with PD. These results provide further insight into how TLR2 activation may promote α-synuclein pathology in PD and support that TLR2 may be a potential therapeutic target for the treatment of PD.
    Keywords:  Parkinson’s disease; fibril; inhibitor; lysosome; synuclein; toll-like receptor
    DOI:  https://doi.org/10.1016/j.jbc.2022.102260
  35. Commun Biol. 2022 Jul 22. 5(1): 726
    mTOR contributes to endothelium-dependent vasorelaxation by promoting eNOS expression and preventing eNOS uncoupling.
    Yiying Wang, Qiannan Li, Zhiyang Zhang, Kai Peng, Dai-Min Zhang, Qianlu Yang, Anthony G Passerini, Scott I Simon, ChongXiu Sun.
      Clinically used inhibitors of mammalian target of rapamycin (mTOR) negatively impacts endothelial-dependent vasodilatation (EDD) through unidentified mechanisms. Here we show that either the endothelium-specific deletion of Mtor to inhibit both mTOR complexes, or depletion of Raptor or Rictor to disrupt mTORC1 or mTORC2, causes impaired EDD, accompanied by reduced NO in the serum of mice. Consistently, inhibition of mTOR decreases NO production by human and mouse EC. Specifically, inhibition of mTORC1 suppresses eNOS gene expression, due to impairment in p70S6K-mediated posttranscriptional regulation of the transcription factor KLF2 expression. In contrast to mTORC1 inhibition, a positive-feedback between MAPK (p38 and JNK) activation and Nox2 upregulation contributes to the excessive generation of reactive oxygen species (ROS), which causes eNOS uncoupling and decreased NO bioavailability in mTORC2-inhibited EC. Adeno-associated virus-mediated EC-specific overexpression of KLF2 or suppression of Nox2 restores EDD function in endothelial mTORC1- or mTORC2-inhibited mice.
    DOI:  https://doi.org/10.1038/s42003-022-03653-w
  36. Cancer Res. 2022 Jul 22. pii: CAN-22-0121. [Epub ahead of print]
    Activating mTOR mutations are detrimental in nutrient-poor conditions.
    Agata A Bielska, Caitlin F Harrigan, Yeon Ju Kyung, Quaid Morris, Wilhelm Palm, Craig B Thompson.
      The mechanistic target of rapamycin (mTOR) is a key regulator of cell growth that integrates growth factor signaling and nutrient availability and is a downstream effector of oncogenic receptor tyrosine kinases (RTKs) and PI3K/Akt signaling. Thus, activating mTOR mutations would be expected to enhance growth in many tumor types. However, tumor sequencing data has shown that mTOR mutations are enriched only in renal clear cell carcinoma, a clinically hypervascular tumor unlikely to be constrained by nutrient availability. To further define this cancer type-specific restriction, we studied activating mutations in mTOR. All mTOR mutants tested enhanced growth in a cell type agnostic manner under nutrient-replete conditions but were detrimental to cell survival in nutrient-poor conditions. Consistently, analysis of tumor data demonstrated that oncogenic mutations in the nutrient-sensing arm of the mTOR pathway display a similar phenotype and were exceedingly rare in human cancers of all types. Together, these data suggest that maintaining the ability to turn off mTOR signaling in response to changing nutrient availability is retained in most naturally occurring tumors.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-22-0121
  37. Exploration (Beijing). 2022 Jun;pii: 20210215. [Epub ahead of print]2(3):
    Super-resolution analyzing spatial organization of lysosomes with an organic fluorescent probe.
    Lei Wang, Rui Chen, Guanqun Han, Xuan Liu, Taosheng Huang, Jiajie Diao, Yujie Sun.
      Lysosomes are multifunctional organelles involved in macromolecule degradation, nutrient sensing and autophagy. Live imaging has revealed lysosome subpopulations with dynamics and characteristic cellular localization. An as-yet unanswered question is whether lysosomes are spatially organized to coordinate and integrate their functions. Combined with super-resolution microscopy, we designed a small organic fluorescent probe, TPAE, that targeted lysosomes with a large Stokes shift. When we analyzed the spatial organization of lysosomes against mitochondria in different cell lines with this probe, we discovered different distance distribution patterns between lysosomes and mitochondria during increased autophagy flux. By using SLC25A46 mutation fibroblasts derived from patients containing highly fused mitochondria with low oxidative phosphorylation, we concluded that unhealthy mitochondria redistributed the subcellular localization of lysosomes, which implies a strong connection between mitochondria and lysosomes.
    Keywords:  Lysosomes; fibroblasts; mitochondria; spatial organization; super resolution microscopy
    DOI:  https://doi.org/10.1002/exp.20210215
  38. Proc Natl Acad Sci U S A. 2022 Jul 26. 119(30): e2201089119
    Reversible mitophagy drives metabolic suppression in diapausing beetles.
    Jacqueline E Lebenzon, Peter W Denezis, Lamees Mohammad, Katherine E Mathers, Kurtis F Turnbull, James F Staples, Brent J Sinclair.
      Many insects enter a state of dormancy (diapause) during winter in which they lower their metabolism to save energy. Metabolic suppression is a hallmark of diapause, yet we know little about the mechanisms underpinning metabolic suppression in winter or how it is reversed in the spring. Here, we show that metabolic suppression in dormant Colorado potato beetles results from the breakdown of flight muscle mitochondria via mitophagy. Diapausing Colorado potato beetles suppress their metabolism by 90%, and this lowered metabolic rate coincides with a similar reduction in flight muscle mitochondrial function and density. During early diapause, beetles increase the expression of mitophagy-related transcripts (Parkin and ATG5) in their flight muscle coincident with an increase in mitophagy-related structures in the flight muscle. Knocking down Parkin expression with RNA interference in diapausing beetles prevented some mitochondrial breakdown and partially restored the whole animal metabolic rate, suggesting that metabolic suppression in diapausing beetles is driven by mitophagy. In other animals and in models of disease, such large-scale mitochondrial degradation is irreversible. However, we show that as diapause ends, beetles reverse mitophagy and increase the expression of PGC1α and NRF1 to replenish flight muscle mitochondrial pools. This mitochondrial biogenesis is activated in anticipation of diapause termination and in the absence of external stimuli. Our study provides a mechanistic link between mitochondrial degradation in insect tissues over the winter and whole-animal metabolic suppression.
    Keywords:  dormancy; insect flight muscle; mitochondria; mitophagy
    DOI:  https://doi.org/10.1073/pnas.2201089119
  39. Sci Signal. 2022 Jul 19. 15(743): eabl9169
    A CRISPR screen targeting PI3K effectors identifies RASA3 as a negative regulator of LFA-1-mediated adhesion in T cells.
    Kristoffer H Johansen, Dominic P Golec, Bonnie Huang, Chung Park, Julie H Thomsen, Silvia Preite, Jennifer L Cannons, Fabien Garçon, Edward C Schrom, Christina J F Courrèges, Tibor Z Veres, James Harrison, Meritxell Nus, James D Phelan, Wolfgang Bergmeier, John H Kehrl, Klaus Okkenhaug, Pamela L Schwartzberg.
      The integrin lymphocyte function-associated antigen 1 (LFA-1) helps to coordinate the migration, adhesion, and activation of T cells through interactions with intercellular adhesion molecule 1 (ICAM-1) and ICAM-2. LFA-1 is activated during the engagement of chemokine receptors and the T cell receptor (TCR) through inside-out signaling, a process that is partially mediated by phosphoinositide 3-kinase (PI3K) and its product phosphatidylinositol 3,4,5-trisphosphate (PIP3). To evaluate potential roles of PI3K in LFA-1 activation, we designed a library of CRISPR/single guide RNAs targeting known and potential PIP3-binding proteins and screened for effects on the ability of primary mouse T cells to bind to ICAM-1. We identified multiple proteins that regulated the binding of LFA-1 to ICAM-1, including the Rap1 and Ras GTPase-activating protein RASA3. We found that RASA3 suppressed LFA-1 activation in T cells, that its expression was rapidly reduced upon T cell activation, and that its activity was inhibited by PI3K. Loss of RASA3 in T cells led to increased Rap1 activation, defective lymph node entry and egress, and impaired responses to T-dependent immunization in mice. Our results reveal a critical role for RASA3 in T cell migration, homeostasis, and function.
    DOI:  https://doi.org/10.1126/scisignal.abl9169
  40. Sci Adv. 2022 Jul 08. 8(27): eabl8809
    Alzheimer's disease: Ablating single master site abolishes tau hyperphosphorylation.
    Kristie Stefanoska, Mehul Gajwani, Amanda R P Tan, Holly I Ahel, Prita R Asih, Alexander Volkerling, Anne Poljak, Arne Ittner.
      Hyperphosphorylation of the neuronal tau protein is a hallmark of neurodegenerative tauopathies such as Alzheimer's disease. A central unanswered question is why tau becomes progressively hyperphosphorylated. Here, we show that tau phosphorylation is governed by interdependence- a mechanistic link between initial site-specific and subsequent multi-site phosphorylation. Systematic assessment of site interdependence identified distinct residues (threonine-50, threonine-69, and threonine-181) as master sites that determine propagation of phosphorylation at multiple epitopes. CRISPR point mutation and expression of human tau in Alzheimer's mice showed that site interdependence governs physiologic and amyloid-associated multi-site phosphorylation and cognitive deficits, respectively. Combined targeting of master sites and p38α, the most central tau kinase linked to interdependence, synergistically ablated hyperphosphorylation. In summary, our work delineates how complex tau phosphorylation arises to inform therapeutic and biomarker design for tauopathies.
    DOI:  https://doi.org/10.1126/sciadv.abl8809
  41. Life Sci. 2022 Jul 13. pii: S0024-3205(22)00506-9. [Epub ahead of print] 120806
    The Rab GTPase in the heart: Pivotal roles in development and disease.
    Jiayi Liu, Xuanjun Zheng, Xiaoqian Wu.
      Rab proteins are a family of small GTPases that function as molecular switches of intracellular vesicle formation and membrane trafficking. As a key factor, Rab GTPase participates in autophagy and protein transport and acts as the central hub of membrane trafficking in eukaryotes. The role of Rab GTPase in neurodegenerative disorders, such as Alzheimer's and Parkinson's, has been extensively investigated; however, its implication in cardiovascular embryogenesis and diseases remains largely unknown. In this review, we summarize previous findings and reveal their importance in the onset and progression of cardiac diseases, as well as their emergence as potential therapeutic targets for cardiovascular disease.
    Keywords:  Autophagy; Cardiac disease; Membrane trafficking; Rab GTPase; Transport; Vesicle
    DOI:  https://doi.org/10.1016/j.lfs.2022.120806
  42. Mitochondrion. 2022 Jul 13. pii: S1567-7249(22)00059-9. [Epub ahead of print]
    The interplay of epilepsy with impaired mitophagy and autophagy linked dementia (MAD): A review of therapeutic approaches.
    Siva Prasad Panda, Yogita Dhurandhar, Mehak Agrawal.
      The duration and, age of dementia have been linked to a higher risk of seizures. The exact mechanism that drives epileptogenesis in impaired mitophagy and autophagy linked dementia (MAD) is fully defined after reviewing the Scopus, Publon, and Pubmed databases. The epileptogenesis in patients with Alzheimer's disease dementia (ADD) and Parkinson's disease dementia (PDD) is due to involvement of amyloid plaques (Aβ), phosphorylated tau (pTau), Parkin, NF-kB and NLRP3 inflammasome.Microglia, the prime protective and inflammatory cells in the brain exert crosstalk between mitophagy and inflammation. Several researchers believed that the inflammatory brain cells microglia could be a therapeutic target for the treatment of a MAD associated epilepsy. There are conventional antiepileptic drugs such as gabapentin, lamotrigine, phenytoin sodium, carbamazepine, oxcarbazepine, felbamate, lamotrigine, valproate sodium, and topiramate are prescribed by a psychiatrist to suppress seizure frequency. Also, the conventional drugs generate serious adverse effects and synergises dementia characteristics. The adverse effect of carbamazepine is neurotoxic and also, damages haemopoietic system and respiratory tract. The phenytoin treatment causes cerebellar defect and anemia. Dementia and epilepsy have a complicated relationship, thus targeting mitophagy for cure of epileptic dementia makes sense. Complementary and alternative medicine (CAM) is one of the rising strategies by many patients of the world, not only to suppress seizure frequency but also to mitigate dementia characteristics of patients. Therefore our present review focus on the interplay between epilepsy and MAD and their treatment with CAM approaches.
    Keywords:  CAM; Epileptogenesis; MAD; NF-kB; NLRP3 inflammasome; Parkin
    DOI:  https://doi.org/10.1016/j.mito.2022.07.002
  43. Cell Death Dis. 2022 Jul 21. 13(7): 634
    FUNDC1-mediated mitophagy and HIF1α activation drives pulmonary hypertension during hypoxia.
    Ruxia Liu, Chunling Xu, Weilin Zhang, Yangpo Cao, Jingjing Ye, Bo Li, Shi Jia, Lin Weng, Yingying Liu, Lei Liu, Ming Zheng.
      Hypoxic pulmonary hypertension (PH) is a progressive disease characterized by hyper-proliferation of pulmonary vascular cells including pulmonary artery smooth muscle cells (PASMCs) and can lead to right heart failure and early death. Selective degradation of mitochondria by mitophagy during hypoxia regulates mitochondrial functions in many cells, however, it is not clear if mitophagy is involved in the pathogenesis of hypoxic PH. By employing the hypoxic mitophagy receptor Fundc1 knockout (KO) and transgenic (TG) mouse models, combined hypoxic PH models, the current study found that mitophagy is actively involved in hypoxic PH through regulating PASMC proliferation. In the pulmonary artery medium from hypoxic PH mice, mitophagy was upregulated, accompanied with the increased active form of FUNDC1 protein and the enhanced binding affinity of FUNDC1 with LC3B. In PASMCs, overexpression of FUNDC1 increased mitophagy and cell proliferation while knockdown of FUNDC1 inhibited hypoxia-induced mitophagy and PASMC proliferation. Stimulation of mitophagy by FUNDC1 in PASMCs elevated ROS production and inhibited ubiquitination of hypoxia inducible factor 1α (HIF1α), and inhibition of mitophagy by FUNDC1 knockdown or knockout abolished hypoxia-induced ROS-HIF1α upregulation. Moreover, Fundc1 TG mice developed severe hemodynamics changes and pulmonary vascular remodeling, and Fundc1 KO mice were much resistant to hypoxic PH. In addition, intraperitoneal injection of a specific FUNDC1 peptide inhibitor to block mitophagy ameliorated hypoxic PH. Our results reveal that during hypoxic PH, FUNDC1-mediated mitophagy is upregulated which activates ROS-HIF1α pathway and promotes PASMC proliferation, ultimately leads to pulmonary vascular remodeling and PH.
    DOI:  https://doi.org/10.1038/s41419-022-05091-2
  44. Open Life Sci. 2022 ;17(1): 710-725
    MT-12 inhibits the proliferation of bladder cells in vitro and in vivo by enhancing autophagy through mitochondrial dysfunction.
    Yan Chen, Chengxing Xia, Chunwei Ye, Feineng Liu, Yitian Ou, Ruping Yan, Haifeng Wang, Delin Yang.
      Bladder cancer (BC) is one of the most common malignancies involving the urinary system. Our previous study demonstrated that cobra venom membrane toxin 12 (MT-12) could effectively inhibit BC cell growth and metastasis and induce apoptosis. However, the specific molecular mechanism remains unknown. In this study, we explored whether MT-12 inhibits BC cell proliferation by inducing autophagy cell death through mitochondrial dysfunction. As a result, MT-12 inhibited proliferation and colony formation in RT4 and T24 cells. In the BC xenograft mouse model, autophagy inhibitor 3-MA alleviated the inhibitory effect of MT-12 on tumor growth. In addition, immunostaining revealed downregulated autophagy in MT-12-treated RT4 and T24 cells. We also found that MT-12 led to dysfunctional mitochondria with decreased mitochondrial membrane potential, mtDNA abundance, and increased ROS production, ultimately inducing autophagic apoptosis via the ROS/JNK/P53 pathway. MT-12 inhibits BC proliferation in vitro and in vivo by enhancing autophagy. MT-12 induces mitochondrial dysfunction and decreases autophagy, leading to increased ROS production, which in turn activates the JNK/p53 pathway, leading to BC apoptosis.
    Keywords:  Cobra venom membrane toxin 12; ROS; autophagy; bladder cancer; mitochondrial dysfunction
    DOI:  https://doi.org/10.1515/biol-2022-0082
  45. Mol Brain. 2022 Jul 18. 15(1): 61
    High glucose and palmitic acid induces neuronal senescence by NRSF/REST elevation and the subsequent mTOR-related autophagy suppression.
    Wen-Jiao Xue, Cheng-Feng He, Ren-Yuan Zhou, Xiao-Die Xu, Lv-Xuan Xiang, Jian-Tao Wang, Xin-Ru Wang, Hou-Guang Zhou, Jing-Chun Guo.
      Cell senescence is a basic aging mechanism. Previous studies have found that the cellular senescence in adipose tissue and other tissues, such as the pancreas, muscle and liver, is associated with the pathogenesis and progression of type 2 diabetes; however, strong evidence of whether diabetes directly causes neuronal senescence in the brain is still lacking. In this study, we constructed a high glucose and palmitic acid (HGP) environment on PC12 neuronal cells and primary mouse cortical neurons to simulate diabetes. Our results showed that after HGP exposure, neurons exhibited obvious senescence-like phenotypes, including increased NRSF/REST level, mTOR activation and cell autophagy suppression. Downregulation of NRSF/REST could remarkably alleviate p16, p21 and γH2A.X upregulations induced by HGP treatment, and enhance mTOR-autophagy of neurons. Our results suggested that the diabetic condition could directly induce neuronal senescence, which is mediated by the upregulation of NRSF/REST and subsequent reduction of mTOR-autophagy.
    DOI:  https://doi.org/10.1186/s13041-022-00947-2
  46. Front Endocrinol (Lausanne). 2022 ;13 898634
    Autophagy in Bone Remodeling: A Regulator of Oxidative Stress.
    Chenyu Zhu, Shiwei Shen, Shihua Zhang, Mei Huang, Lan Zhang, Xi Chen.
      Bone homeostasis involves bone formation and bone resorption, which are processes that maintain skeletal health. Oxidative stress is an independent risk factor, causing the dysfunction of bone homeostasis including osteoblast-induced osteogenesis and osteoclast-induced osteoclastogenesis, thereby leading to bone-related diseases, especially osteoporosis. Autophagy is the main cellular stress response system for the limination of damaged organelles and proteins, and it plays a critical role in the differentiation, apoptosis, and survival of bone cells, including bone marrow stem cells (BMSCs), osteoblasts, osteoclasts, and osteocytes. High evels of reactive oxygen species (ROS) induced by oxidative stress induce autophagy to protect against cell damage or even apoptosis. Additionally, pathways such as ROS/FOXO3, ROS/AMPK, ROS/Akt/mTOR, and ROS/JNK/c-Jun are involved in the regulation of oxidative stress-induced autophagy in bone cells, including osteoblasts, osteocytes and osteoclasts. This review discusses how autophagy regulates bone formation and bone resorption following oxidative stress and summarizes the potential protective mechanisms exerted by autophagy, thereby providing new insights regarding bone remodeling and potential therapeutic targets for osteoporosis.
    Keywords:  autophagy; osteoblast; osteoclast; osteoporosis; oxidative stress
    DOI:  https://doi.org/10.3389/fendo.2022.898634
  47. J Cardiovasc Pharmacol. 2022 Jul 18.
    Atorvastatin ameliorates doxorubicin-induced cardiomyopathy by regulating the autophagy-lysosome pathway and its upstream regulatory factor transcription factor EB.
    Shiliang Jiang, Wan-Ching Chou, Linqing Tao, Zhaoyang Qiu, Ge Gao.
       ABSTRACT: Doxorubicin (DOX) is a widely used anticancer drug in clinical practice, and its myocardial toxicity is the main concern in oncotherapy. Statins are commonly used as hypolipidemic drugs. Recent studies have also focused on the effects of statins on autophagy. Autophagy is a process in which cells consume their own cytoplasm or organelles after stimulation, and finally degrade the phagosome in the lysosome. Transcription factor EB (TFEB) is the main factor regulating lysosomal gene transcription and function. We found that atorvastatin increased TFEB protein levels and the ratio of LAMP2/LC3B in the myocardial tissue of mice with doxorubicin-induced cardiomyopathy (DIC). Therefore, we speculated that atorvastatin may improve cardiac function in mice with DIC by increasing the expression of TFEB to enhance lysosomal function and autophagy. This study explored the role of TFEB in DIC and the possible mechanism of atorvastatin in improving DIC and used statins to prevent and treat DIC; various dilated cardiomyopathy and heart failure diseases provide more experimental evidence. All relevant data are within the paper and its Supporting Information files.
    DOI:  https://doi.org/10.1097/FJC.0000000000001334
  48. Front Oncol. 2022 ;12 915240
    RAC3 Inhibition Induces Autophagy to Impair Metastasis in Bladder Cancer Cells via the PI3K/AKT/mTOR Pathway.
    Liwei Wang, Jiazhong Shi, Sha Liu, Yaqin Huang, Hua Ding, Baixiong Zhao, Yuting Liu, Wuxing Wang, Jin Yang, Zhiwen Chen.
       Background: Bladder cancer (BCa) is one of the most frequent malignant tumors globally, with a significant morbidity and mortality rate. Gene expression dysregulation has been proven to play a critical role in tumorigenesis. Ras-related C3 botulinum toxin substrate3 (RAC3), which is overexpressed in several malignancies and promotes tumor progression, has been identified as an oncogene. However, RAC3 has important but not fully understood biological functions in cancer. Our research aims to reveal the new functions and potential mechanisms of RAC3 involved in BCa progression.
    Methods: We explored the expression level of RAC3 and its relationship with prognosis by publicly accessible BCa datasets, while the correlation of RAC3 expression with clinicopathological variables of patients was analyzed. In vitro and in vivo proliferation, migration, autophagy, and other phenotypic changes were examined by constructing knockdown(KD)/overexpression(OE) RAC3 cells and their association with PI3K/AKT/mTOR pathway was explored by adding autophagy-related compounds.
    Results: Compared with non-tumor samples, RAC3 was highly expressed in BCa and negatively correlated with prognosis. KD/OE RAC3 inhibited/promoted the proliferation and migration of BCa cells. Knockdown RAC3 caused cell cycle arrest and decreased adhesion without affecting apoptosis. Inhibition of RAC3 activates PI3K/AKT/mTOR mediated autophagy and inhibits proliferation and migration of BCa cells in vivo and in vitro. Autophagy inhibitor 3MA can partially rescue the metastasis and proliferation inhibition effect caused by RAC3 inhibition. Inhibit/activate mTOR enhanced/impaired autophagy, resulting in shRAC3-mediated migration defect exacerbated/rescued.
    Conclusion: RAC3 is highly expressed in BCa. It is associated with advanced clinicopathological variables and poor prognosis. Knockdown RAC3 exerts an antitumor effect by enhancing PI3K/AKT/mTOR mediated autophagy. Targeting RAC3 and autophagy simultaneously is a potential therapeutic strategy for inhibiting BCa progression and prolonging survival.
    Keywords:  PI3K/AKT/mTOR; RAC3; autophagy; bladder cancer; metastasis
    DOI:  https://doi.org/10.3389/fonc.2022.915240
  49. Biogerontology. 2022 Jul 16.
    Mitochondrial function and nutrient sensing pathways in ageing: enhancing longevity through dietary interventions.
    Elangbam Tomtheelnganbee, Puja Sah, R Sharma.
      Ageing is accompanied by alterations in several biochemical processes, highly influenced by its environment. It is controlled by the interactions at various levels of biological hierarchy. To maintain homeostasis, a number of nutrient sensors respond to the nutritional status of the cell and control its energy metabolism. Mitochondrial physiology is influenced by the energy status of the cell. The alterations in mitochondrial physiology and the network of nutrient sensors result in mitochondrial damage leading to age related metabolic degeneration and diseases. Calorie restriction (CR) has proved to be as the most successful intervention to achieve the goal of longevity and healthspan. CR elicits a hormetic response and regulates metabolism by modulating these networks. In this review, the authors summarize the interdependent relationship between mitochondrial physiology and nutrient sensors during the ageing process and their role in regulating metabolism.
    Keywords:  AMPK; Ageing; Dietary restriction; Mitochondria; Sirtuins; mTOR
    DOI:  https://doi.org/10.1007/s10522-022-09978-7
  50. Sci Adv. 2022 Jul 15. 8(28): eabn3326
    Neuronal activity induces glucosylceramide that is secreted via exosomes for lysosomal degradation in glia.
    Liping Wang, Guang Lin, Zhongyuan Zuo, Yarong Li, Seul Kee Byeon, Akhilesh Pandey, Hugo J Bellen.
      Recessive variants in GBA1 cause Gaucher disease, a prevalent form of lysosome storage disease. GBA1 encodes a lysosomal enzyme that hydrolyzes glucosylceramide (GlcCer) into glucose and ceramide. Its loss causes lysosomal dysfunction and increased levels of GlcCer. We generated a null allele of the Drosophila ortholog Gba1b by inserting the Gal4 using CRISPR-Cas9. Here, we show that Gba1b is expressed in glia but not in neurons. Glial-specific knockdown recapitulates the defects found in Gba1b mutants, and these can be rescued by glial expression of human GBA1. We show that GlcCer is synthesized upon neuronal activity, and it is transported from neurons to glia through exosomes. Furthermore, we found that glial TGF-β/BMP induces the transfer of GlcCer from neurons to glia and that the White protein, an ABCG transporter, promotes GlcCer trafficking to glial lysosomes for degradation.
    DOI:  https://doi.org/10.1126/sciadv.abn3326
  51. Nat Cell Biol. 2022 Jul 18.
    ADAR1 downregulation by autophagy drives senescence independently of RNA editing by enhancing p16INK4a levels.
    Xue Hao, Yusuke Shiromoto, Masayuki Sakurai, Martina Towers, Qiang Zhang, Shuai Wu, Aaron Havas, Lu Wang, Shelley Berger, Peter D Adams, Bin Tian, Kazuko Nishikura, Andrew V Kossenkov, Pingyu Liu, Rugang Zhang.
      Cellular senescence plays a causal role in ageing and, in mice, depletion of p16INK4a-expressing senescent cells delays ageing-associated disorders1,2. Adenosine deaminases acting on RNA (ADARs) are RNA-editing enzymes that are also implicated as important regulators of human ageing, and ADAR inactivation causes age-associated pathologies such as neurodegeneration in model organisms3,4. However, the role, if any, of ADARs in cellular senescence is unknown. Here we show that ADAR1 is post-transcriptionally downregulated by autophagic degradation to promote senescence through p16INK4a upregulation. The ADAR1 downregulation is sufficient to drive senescence in both in vitro and in vivo models. Senescence induced by ADAR1 downregulation is p16INK4a-dependent and independent of its RNA-editing function. Mechanistically, ADAR1 promotes SIRT1 expression by affecting its RNA stability through HuR, an RNA-binding protein that increases the half-life and steady-state levels of its target mRNAs. SIRT1 in turn antagonizes translation of mRNA encoding p16INK4a. Hence, downregulation of ADAR1 and SIRT1 mediates p16INK4a upregulation by enhancing its mRNA translation. Finally, Adar1 is downregulated during ageing of mouse tissues such as brain, ovary and intestine, and Adar1 expression correlates with Sirt1 expression in these tissues in mice. Together, our study reveals an RNA-editing-independent role for ADAR1 in the regulation of senescence by post-transcriptionally controlling p16INK4a expression.
    DOI:  https://doi.org/10.1038/s41556-022-00959-z
  52. Mutat Res. 2022 Jul 09. pii: S0027-5107(22)00017-3. [Epub ahead of print]825 111790
    The kidney-expressed transcription factor ZKSCAN3 is dispensable for autophagy transcriptional regulation and AKI progression in mouse.
    Zejian Liu, Xiaoyu Li, Xingyu Li, Zixian Li, Huixia Chen, Siqiao Gong, Minjie Zhang, Yaozhi Zhang, Zhihang Li, Lin Yang, Huafeng Liu.
      Acute kidney injury (AKI) is a common clinical disease that can cause serious harm to the kidneys, but it has no effective treatment till now. The modulation of autophagy pathway regulation is considered a potentially effective therapeutic approach in AKI prevention and treatment. ZKSCAN3 has been shown to be an important transcription factor that negatively regulates autophagy activity in cancer tissues. In order to determine whether autophagy could be activated by knocking out ZKSCAN3 to exert the renal protective effect of autophagy, we constructed AKI models with Zkscan3 knockout (KO) mice and detected renal pathological changes and renal function changes as well as autophagy-related indicators. We found that Zkscan3 KO had no significant effect on kidney development. Besides, no significant changes in autophagy activity were observed under normal physiological or AKI conditions. In non-tumor tissues, ZKSCAN3 did not mediate transcriptional regulation of autophagy-related genes. These findings suggest that because ZKSCAN3 may not function in the transcriptional regulation of autophagy-related genes in non-tumor tissues, it may not be used as a therapeutic target for AKI.
    Keywords:  Acute kidney injury; Autophagy; Cisplatin; Sepsis; ZKSCAN3
    DOI:  https://doi.org/10.1016/j.mrfmmm.2022.111790
This page is being maintained by Thomas Krichel. It was last updated on 2024‒11‒22 at 4:11.