bims-auttor 2021-10-17 papers

bims-auttor

Biomed News

on Autophagy and mTOR

Issue of 2021–10–17
43 papers selected by
Viktor Korolchuk, Newcastle University

A new type of membrane contact in the ER-Golgi system regulates autophagosome biogenesis.
The protein N-terminal acetyltransferase A complex contributes to yeast mitophagy via promoting expression and phosphorylation of Atg32.
ULK1 promotes mitophagy via phosphorylation and stabilization of BNIP3.
Phase separation of Nur77 mediates celastrol-induced mitophagy by promoting the liquidity of p62/SQSTM1 condensates.
Chaperone-mediated autophagy and disease: Implications for cancer and neurodegeneration.
Human autophagy measurement: an underappreciated barrier to translation.
EIF4A3: a gatekeeper of autophagy.
Drosophila Rab39 Attenuates Lysosomal Degradation.
Lactate and Pyruvate Activate Autophagy and Mitophagy that Protect Cells in Toxic Model of Parkinson's Disease.
Impaired Mitophagy in Neurons and Glial Cells during Aging and Age-Related Disorders.
Loss-of-function mutation in VCP mimics the characteristic pathology as in FTLD-TARDBP.
Nicotinamide Riboside and Metformin Ameliorate Mitophagy Defect in Induced Pluripotent Stem Cell-Derived Astrocytes With POLG Mutations.
DYNC1LI2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and improves cellular homeostasis in cystinosis.
The necessity of nucleophagic modality.
Functional heterogeneity of POMC neurons relies on mTORC1 signaling.
The selective degradation of sirtuins via macroautophagy in the MPP+ model of Parkinson's disease is promoted by conserved oxidation sites.
Cellular senescence-an aging hallmark in chronic obstructive pulmonary disease pathogenesis.
Activation of the purinergic receptor P2X7 improves hepatosteatosis by promoting lipophagy.
PGRMC1 acts as a size-selective cargo receptor to drive ER-phagic clearance of mutant prohormones.
Prefused lysosomes cluster on autophagosomes regulated by VAMP8.
CHD1L augments autophagy-mediated migration of hepatocellular carcinoma through targeting ZKSCAN3.
Tyrosine Phosphorylation of DEPTOR Functions as a Molecular Switch to Activate mTOR Signaling.
How Oncogenic Viruses Exploit p62-Mediated Selective Autophagy for Cancer Development.
Autophagy and cancer metabolism-The two-way interplay.
Mitophagy in Diabetic Cardiomyopathy: Roles and Mechanisms.
Epilepsy in the mTORopathies: opportunities for precision medicine.
USP30-AS1 contributes to mitochondrial quality control in glioblastoma cells.
A leucine-derived fatty acid unlocks the mTOR to development.
Defective lipid droplet - lysosome interaction causes fatty liver disease as evidenced by human mutations in TMEM199 and CCDC115.
Perspective on mTOR-dependent Protection in Status Epilepticus.
mTOR-dependent immunometabolism as Achilles' heel of anticancer therapy.
Targeting Mitochondrial Network Disorganization is Protective in C. elegans Models of Huntington's Disease.
Metformin may induce ferroptosis by inhibiting autophagy via lncRNA H19 in breast cancer.
GluN2A-ERK-mTOR pathway confers a vulnerability to LPS-induced depressive-like behaviour.
OAB-14 Effectively Ameliorates the Dysfunction of the Endosomal-Autophagic-Lysosomal Pathway in APP/PS1 Transgenic Mice.
Sestrin2-Mediated Autophagy Contributes to Drug Resistance via Endoplasmic Reticulum Stress in Human Osteosarcoma.
Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies.
IFN-γ mediates Paneth cell death via suppression of mTOR.
Regulation of electrogenic Na+ /HCO3 - cotransporter 1 (NBCe1) function and its dependence on m-TOR mediated phosphorylation of Ser245.
Therapeutic potential of PGC-1α in age-related macular degeneration (AMD) - the involvement of mitochondrial quality control, autophagy, and antioxidant response.
The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation.
Cholecystokinin 1 receptor activation restores normal mTORC1 signaling and is protective to Purkinje cells of SCA mice.
Autophagy in skin barrier and immune-related skin diseases.

Autophagy. 2021 Oct 13. 1-3

A new type of membrane contact in the ER-Golgi system regulates autophagosome biogenesis.

Shulin Li, Min Zhang, Liang Ge.

  Formation of the double-membrane autophagosome requires membrane reorganization of the endomembrane system to generate membrane precursors. The ER-Golgi trafficking system has been shown to provide membranes for phagophore growth. Nonetheless, how the components of the ER-Golgi system are redirected toward autophagosome biogenesis remains unclear. Here, we identify a new type of membrane contact formed between the ER-Golgi intermediate compartment (ERGIC) and the ER-exit sites (ERES) under macroautophagy/autophagy-induction conditions. The ERGIC-ERES contact is established by the TMED9-PREB/SEC12 interaction and regulates the biogenesis of the ERGIC-COPII vesicles, which we found previously act as a membrane template for LC3 lipidation and autophagosome formation.

Keywords:  Autophagosome; COPII; ERES; ERGIC; SEC12; TMED9; autophagy

DOI:  https://doi.org/10.1080/15548627.2021.1972406
J Biochem. 2021 Oct 11. 170(2): 175-182

The protein N-terminal acetyltransferase A complex contributes to yeast mitophagy via promoting expression and phosphorylation of Atg32.

Mitsutaka Kubota, Koji Okamoto.

  Mitophagy is an evolutionarily conserved catabolic process that selectively degrades damaged or superfluous mitochondria via autophagy. Although mitophagy is considered to be critical to maintain cellular homeostasis, detailed mechanisms of mitophagy remain largely unknown. In the budding yeast Saccharomyces cerevisiae, the protein N-terminal acetyltransferase A (NatA) complex is important for transcriptional induction of the pro-mitophagic factor Atg32 and efficient degradation of mitochondria under prolonged respiratory conditions. Overexpression of Atg32 only partially recovers mitophagy in cells lacking NatA, raising the possibility that NatA may contribute to mitophagy via additional mechanisms. Here, we demonstrate that Atg32 phosphorylation, which is required for facilitating mitophagy, is altered in respiring NatA-deficient cells. Hyperphosphorylation of Atg32 partially rescues mitophagy in cells lacking NatA. Notably, mitophagy is mostly restored in NatA-null cells overexpressing hyperphosphorylated Atg32. Loss of NatA does not impair the interaction of phosphorylated Atg32 with Atg11, a scaffold protein critical for selective autophagy, suggesting that NatA-dependent Atg32 phosphorylation promotes mitophagy independently of Atg32-Atg11 interactions. We propose that NatA-mediated protein N-terminal acetylation acts in Atg32 expression and phosphorylation to drive mitophagy.

Keywords:  Atg32; NatA; Ppg1; autophagy; mitochondria; yeast

DOI:  https://doi.org/10.1093/jb/mvab068
Sci Rep. 2021 Oct 15. 11(1): 20526

ULK1 promotes mitophagy via phosphorylation and stabilization of BNIP3.

Logan P Poole, Althea Bock-Hughes, Damian E Berardi, Kay F Macleod.

UNC51-like kinase-1 (ULK1) is the catalytic component of the autophagy pre-initiation complex that stimulates autophagy via phosphorylation of ATG14, BECLN1 and other autophagy proteins. ULK1 has also been shown to specifically promote mitophagy but the mechanistic basis of how has remained unclear. Here we show that ULK1 phosphorylates the BNIP3 mitochondrial cargo receptor on a critical serine residue (S17) adjacent to its amino terminal LIR motif. ULK1 similarly phosphorylates BNIP3L on S35. Phosphorylation of BNIP3 on S17 by ULK1 promotes interaction with LC3 and mitophagy. ULK1 interaction also promotes BNIP3 protein stability by limiting its turnover at the proteasome. The ability of ULK1 to regulate BNIP3 protein stability depends on an intact "BH3" domain and deletion of its "BH3" domain reduces BNIP3 turnover and increases BNIP3 protein levels independent of ULK1. In summary ULK1 promotes mitophagy by both stabilization of BNIP3 protein and via phosphorylation of S17 to stimulate interaction with LC3.

DOI: https://doi.org/10.1038/s41598-021-00170-4
Nat Commun. 2021 Oct 13. 12(1): 5989

Phase separation of Nur77 mediates celastrol-induced mitophagy by promoting the liquidity of p62/SQSTM1 condensates.

Shuang-Zhou Peng, Xiao-Hui Chen, Si-Jie Chen, Jie Zhang, Chuan-Ying Wang, Wei-Rong Liu, Duo Zhang, Ying Su, Xiao-Kun Zhang.

Liquid-liquid phase separation promotes the formation of membraneless condensates that mediate diverse cellular functions, including autophagy of misfolded proteins. However, how phase separation participates in autophagy of dysfunctional mitochondria (mitophagy) remains obscure. We previously discovered that nuclear receptor Nur77 (also called TR3, NGFI-B, or NR4A1) translocates from the nucleus to mitochondria to mediate celastrol-induced mitophagy through interaction with p62/SQSTM1. Here, we show that the ubiquitinated mitochondrial Nur77 forms membraneless condensates capable of sequestrating damaged mitochondria by interacting with the UBA domain of p62/SQSTM1. However, tethering clustered mitochondria to the autophagy machinery requires an additional interaction mediated by the N-terminal intrinsically disordered region (IDR) of Nur77 and the N-terminal PB1 domain of p62/SQSTM1, which confers Nur77-p62/SQSTM1 condensates with the magnitude and liquidity. Our results demonstrate how composite multivalent interaction between Nur77 and p62/SQSTM1 coordinates to sequester damaged mitochondria and to connect targeted cargo mitochondria for autophagy, providing mechanistic insight into mitophagy.

DOI: https://doi.org/10.1038/s41467-021-26295-8
Mol Aspects Med. 2021 Oct 07. pii: S0098-2997(21)00085-6. [Epub ahead of print] 101025

Chaperone-mediated autophagy and disease: Implications for cancer and neurodegeneration.

Raquel Gómez-Sintes, Esperanza Arias.

  Chaperone-mediated autophagy (CMA) is a proteolytic process whereby selected intracellular proteins are degraded inside lysosomes. Owing to its selectivity, CMA participates in the modulation of specific regulatory proteins, thereby playing an important role in multiple cellular processes. Studies conducted over the last two decades have enabled the molecular characterization of this autophagic pathway and the design of specific experimental models, and have underscored the importance of CMA in a range of physiological processes beyond mere protein quality control. Those findings also indicate that decreases in CMA function with increasing age may contribute to the pathogenesis of age-associated diseases, including neurodegeneration and cancer. In the context of neurological diseases, CMA impairment is thought to contribute to the accumulation of misfolded/aggregated proteins, a process central to the pathogenesis of neurodegenerative diseases. CMA therefore constitutes a potential therapeutic target, as its induction accelerates the clearance of pathogenic proteins, promoting cell survival. More recent evidence has highlighted the important and complex role of CMA in cancer biology. While CMA induction may limit tumor development, experimental evidence also indicates that inhibition of this pathway can attenuate the growth of established tumors and improve the response to cancer therapeutics. Here, we describe and discuss the evidence supporting a role of impaired CMA function in neurodegeneration and cancer, as well as future research directions to evaluate the potential of this pathway as a target for the prevention and treatment of these diseases.

Keywords:  Aggregation; Autophagy; Chaperones; Lysosomes; Protein degradation; Tumorigenesis

DOI:  https://doi.org/10.1016/j.mam.2021.101025
Trends Mol Med. 2021 Oct 07. pii: S1471-4914(21)00251-3. [Epub ahead of print]

Human autophagy measurement: an underappreciated barrier to translation.

Timothy J Sargeant, Julien Bensalem.

  Preclinical research shows that autophagy is a modifiable process that holds promise for preventing human age-related disease. However, this knowledge has not been clinically translated. Here, we discuss recent developments in the ability to measure human autophagy, and why it is a critical step for translation.

Keywords:  ageing; autophagic flux; autophagy; blood; human; translation

DOI:  https://doi.org/10.1016/j.molmed.2021.09.003
Autophagy. 2021 Oct 13. 1-2

EIF4A3: a gatekeeper of autophagy.

Despoina Sakellariou, Lisa B Frankel.

  EIF4A3 (eukaryotic translation initiation factor 4A3) is an RNA helicase and core component of the exon junction complex. While this RNA-binding protein (RBP) is well-characterized for its crucial roles in splicing, RNA trafficking and nonsense-mediated decay, its role in the regulation of metabolic signaling pathways remains elusive. In a recent study, we describe a new role for EIF4A3 as a negative regulator of macroautophagy/autophagy. Mechanistically, we report that EIF4A3, through its ability to safeguard splicing, can maintain low basal levels of autophagy through the cytosolic retention of the key autophagy transcription factor TFEB. Upon EIF4A3 depletion, the shuttling of TFEB to the nucleus results in an integrated transcriptional response, which induces both early and late steps of the autophagy pathway and enhances autophagic flux. We further report the upregulation of EIF4A3 across multiple cancer types and highlight the relevance of this newly identified EIF4A3-TFEB signaling axis in human tumors.

Keywords:  Autophagy regulation; EIF4A3; GSK3B; RNA-binding proteins; TFEB; cancer; exon skipping

DOI:  https://doi.org/10.1080/15548627.2021.1985881
Int J Mol Sci. 2021 Sep 30. pii: 10635. [Epub ahead of print]22(19):

Drosophila Rab39 Attenuates Lysosomal Degradation.

Zsolt Lakatos, Péter Benkő, Gábor Juhász, Péter Lőrincz.

  Lysosomal degradation, the common destination of autophagy and endocytosis, is one of the most important elements of eukaryotic metabolism. The small GTPases Rab39A and B are potential new effectors of this pathway, as their malfunction is implicated in severe human diseases like cancer and neurodegeneration. In this study, the lysosomal regulatory role of the single Drosophila Rab39 ortholog was characterized, providing valuable insight into the potential cell biological mechanisms mediated by these proteins. Using a de novo CRISPR-generated rab39 mutant, we found no failure in the early steps of endocytosis and autophagy. On the contrary, we found that Rab39 mutant nephrocytes internalize and degrade endocytic cargo at a higher rate compared to control cells. In addition, Rab39 mutant fat body cells contain small yet functional autolysosomes without lysosomal fusion defect. Our data identify Drosophila Rab39 as a negative regulator of lysosomal clearance during both endocytosis and autophagy.

Keywords:  Drosophila; Rab39; autophagy; endocytosis; lysosomes

DOI:  https://doi.org/10.3390/ijms221910635
Mol Neurobiol. 2021 Oct 13.

Lactate and Pyruvate Activate Autophagy and Mitophagy that Protect Cells in Toxic Model of Parkinson's Disease.

Evgeniya I Fedotova, Ludmila P Dolgacheva, Andrey Y Abramov, Alexey V Berezhnov.

  Intracellular quality control regulated by autophagy process is important for maintenance of cellular homeostasis. Deregulation of autophagy and more specifically mitophagy leads to accumulation of the misfolded proteins and damaged mitochondria that in turn leads to the cell loss. Alteration of autophagy and mitophagy has shown to be involved in the number of disorders including neurodegenerative diseases. Autophagy and mitophagy could be activated by short-time acidification of the cytosol; however, most of the compounds which can induce it are toxic. Here, we tested several organic compounds which are involved in cellular metabolism on their ability to change intracellular pH and induce mitophagy/autophagy. We have found that lactate and pyruvate are able to reduce intracellular pH in non-toxic concentrations. Short-term (2 h) and long-term (24 h) incubation of the cells with lactate and pyruvateinduced mitophagy and autophagy. Incubation of the SH-SY5Y cells or primary neurons and astrocytes with lactate or pyruvate also activated mitophagy and autophagy after MPP + treatment that led to recovery of mitochondrial function and protection of these cells against apoptotic and necrotic death. Thus, pyruvate- or lactate-induced acidification of cytosol activates cell protective mitophagy and autophagy.

Keywords:  Autophagy; Intracellular pH; MPP + ; Mitophagy; Parkinson’s model; Pyruvate

DOI:  https://doi.org/10.1007/s12035-021-02583-8
Int J Mol Sci. 2021 Sep 23. pii: 10251. [Epub ahead of print]22(19):

Impaired Mitophagy in Neurons and Glial Cells during Aging and Age-Related Disorders.

Vladimir Sukhorukov, Dmitry Voronkov, Tatiana Baranich, Natalia Mudzhiri, Alina Magnaeva, Sergey Illarioshkin.

  Aging is associated with a decline in cognitive function, which can partly be explained by the accumulation of damage to the brain cells over time. Neurons and glia undergo morphological and ultrastructure changes during aging. Over the past several years, it has become evident that at the cellular level, various hallmarks of an aging brain are closely related to mitophagy. The importance of mitochondria quality and quantity control through mitophagy is highlighted by the contribution that defects in mitochondria-autophagy crosstalk make to aging and age-related diseases. In this review, we analyze some of the more recent findings regarding the study of brain aging and neurodegeneration in the context of mitophagy. We discuss the data on the dynamics of selective autophagy in neurons and glial cells during aging and in the course of neurodegeneration, focusing on three mechanisms of mitophagy: non-receptor-mediated mitophagy, receptor-mediated mitophagy, and transcellular mitophagy. We review the role of mitophagy in neuronal/glial homeostasis and in the molecular pathogenesis of neurodegenerative disorders, such as Parkinson's disease, Alzheimer's disease, and other disorders. Common mechanisms of aging and neurodegeneration that are related to different mitophagy pathways provide a number of promising targets for potential therapeutic agents.

Keywords:  Alzheimer’s disease; Parkinson’s disease; aging; mitophagy; neurodegeneration

DOI:  https://doi.org/10.3390/ijms221910251
Autophagy. 2021 Oct 10. 1-2

Loss-of-function mutation in VCP mimics the characteristic pathology as in FTLD-TARDBP.

Abubakar Wani, Conrad C Weihl.

  VCP (valosin containing protein), a member of the AAA+ protein family, is critical for many cellular processes and functions. Dominant VCP mutations cause a rare neurodegenerative disease known as multisystem proteinopathy (MSP). The spectrum of mechanisms causing fronto-temporal dementia with TARDBP/TDP-43 inclusions (FTLD-TARDBP) by VCP disease mutations remains unclear. Our recent work identified VCP activity as a mediator of FTLD-TARDBP. Specifically, brain atrophy, behavioral changes, neuronal loss, gliosis, and TARDBP pathology were observed in vcp conditional knockout (cKO) mice. We also found that autophago-lysosomal dysfunction, TARDBP inclusions, and ubiquitin-proteasome impairment precede neuronal loss. We further studied conditional expression of the disease-associated mutation VCPR155C in vcp-null mice. We observed features similar to those of VCP inactivation, suggesting that VCP mutation is hypomorphic. Furthermore, proteomic, and transcriptomic signatures in vcp cKO mice resemble those of GRN/Progranulin carriers. Therefore, VCP is essential for neuronal survival by several mechanisms and could be a therapeutic target aimed at restoring protein homeostasis in patients with FTLD-TARDBP.

Keywords:  Autophagy; FTLD-TDP-43; neurodegeneration; progranulin; valosin-containing protein

DOI:  https://doi.org/10.1080/15548627.2021.1985880
Front Cell Dev Biol. 2021 ;9 737304

Nicotinamide Riboside and Metformin Ameliorate Mitophagy Defect in Induced Pluripotent Stem Cell-Derived Astrocytes With POLG Mutations.

Anbin Chen, Cecilie Katrin Kristiansen, Yu Hong, Atefeh Kianian, Evandro Fei Fang, Gareth John Sullivan, Jian Wang, Xingang Li, Laurence A Bindoff, Kristina Xiao Liang.

  Mitophagy specifically recognizes and removes damaged or superfluous mitochondria to maintain mitochondrial homeostasis and proper neuronal function. Defective mitophagy and the resulting accumulation of damaged mitochondria occur in several neurodegenerative diseases. Previously, we showed mitochondrial dysfunction in astrocytes with POLG mutations, and here, we examined how POLG mutations affect mitophagy in astrocytes and how this can be ameliorated pharmacologically. Using induced pluripotent stem cell (iPSC)-derived astrocytes carrying POLG mutations, we found downregulation of mitophagy/autophagy-related genes using RNA sequencing-based KEGG metabolic pathway analysis. We confirmed a deficit in mitochondrial autophagosome formation under exogenous stress conditions and downregulation of the mitophagy receptor p62, reduced lipidation of LC3B-II, and decreased expression of lysosome protein lysosomal-associated membrane protein 2A (LAMP2A). These changes were regulated by the PINK1/Parkin pathway and AKT/mTOR/AMPK/ULK1 signaling pathways. Importantly, we found that double treatment with nicotinamide riboside (NR) and metformin rescued mitophagy defects and mitochondrial dysfunction in POLG-mutant astrocytes. Our findings reveal that impaired mitophagy is involved in the observed mitochondrial dysfunction caused by POLG mutations in astrocytes, potentially contributing to the phenotype in POLG-related diseases. This study also demonstrates the therapeutic potential of NR and metformin in these incurable mitochondrial diseases.

Keywords:  IPSC (induced pluripotent stem cells); POLG; astrocytes; metformin; mitochondria; mitophagy; nicotinamide riboside (NR)

DOI:  https://doi.org/10.3389/fcell.2021.737304
Autophagy. 2021 Oct 13. 1-19

DYNC1LI2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and improves cellular homeostasis in cystinosis.

Farhana Rahman, Jennifer L Johnson, Jinzhong Zhang, Jing He, Kersi Pestonjamasp, Stephanie Cherqui, Sergio D Catz.

  The dynein motor protein complex is required for retrograde transport but the functions of the intermediate-light chains that form the cargo-binding complex are not elucidated and the importance of individual subunits in maintaining cellular homeostasis is unknown. Here, using mRNA arrays and protein analysis, we show that the dynein subunit, DYNC1LI2 (dynein, cytoplasmic 1 light intermediate chain 2) is downregulated in cystinosis, a lysosomal storage disorder caused by genetic defects in CTNS (cystinosin, lysosomal cystine transporter). Reconstitution of DYNC1LI2 expression in ctns-/- cells reestablished endolysosomal dynamics. Defective vesicular trafficking in cystinotic cells was rescued by DYNC1LI2 expression which correlated with decreased endoplasmic reticulum stress manifested as decreased expression levels of the chaperone HSPA5/GRP78, and the transcription factors ATF4 and DDIT3/CHOP. Mitochondrial fragmentation, membrane potential and endolysosomal-mitochondrial association in cystinotic cells were rescued by DYNC1LI2. Survival of cystinotic cells to oxidative stress was increased by DYNC1LI2 reconstitution but not by its paralog DYNC1LI1, which also failed to decrease ER stress and mitochondrial fragmentation. DYNC1LI2 expression rescued the localization of the chaperone-mediated autophagy (CMA) receptor LAMP2A, CMA activity, cellular homeostasis and LRP2/megalin expression in cystinotic proximal tubule cells, the primary cell type affected in cystinosis. DYNC1LI2 failed to rescue phenotypes in cystinotic cells when LAMP2A was downregulated or when co-expressed with dominant negative (DN) RAB7 or DN-RAB11, which impaired LAMP2A trafficking. DYNC1LI2 emerges as a regulator of cellular homeostasis and potential target to repair underlying trafficking and CMA in cystinosis, a mechanism that is not restored by lysosomal cystine depletion therapies.Abbreviations: ACTB: actin, beta; ATF4: activating transcription factor 4; CMA: chaperone-mediated autophagy; DYNC1LI1: dynein cytoplasmic 1 light intermediate chain 1; DYNC1LI2: dynein cytoplasmic 1 light intermediate chain 2; ER: endoplasmic reticulum; LAMP1: lysosomal associated membrane protein 1; LAMP2A: lysosomal associated membrane protein 2A; LIC: light-intermediate chains; LRP2/Megalin: LDL receptor related protein 2; PTCs: proximal tubule cells; RAB: RAB, member RAS oncogene family; RAB11FIP3: RAB11 family interacting protein 3; RILP: Rab interacting lysosomal protein.

Keywords:  Lysosomal storage disorder; megalin; proximal tubule cell; rab gtpases; trafficking

DOI:  https://doi.org/10.1080/15548627.2021.1971937
Autophagy. 2021 Oct 13. 1-6

The necessity of nucleophagic modality.

Dalibor Mijaljica, Daniel J Klionsky.

  Nucleophagy, the selective subtype of autophagy that predominantly targets only a selected and (nonessential) portion of the nucleus, and rarely the nucleus in its entirety, for degradation, reinforces the paradigm that nucleophagy recycling is a meticulous and highly delicate process guarded by fail-safe mechanisms. Our goal in this commentary is to encourage autophagy researchers and other scientists to explore nucleophagy blind spots and gain advanced insights into the diverse roles of this process and its selective modality as they pertain to intranuclear quality control and cellular homeostasis. Identifying and deciphering nucleophagic signaling, regulation, molecular mechanism(s) and its mediators, cargo composition and nuclear membrane dynamics under numerous physiological and/or pathological settings will provide important advances in our understanding of this critical type of organelle-selective autophagy.Abbreviations: INM, inner nuclear membrane; LN, late nucleophagy; mRNA, messenger RNA; NE, nuclear envelope; NL, nuclear lamina; NPC(s), nuclear pore complex(es); NVJ(s), nucleus-vacuole junction(s); ONM, outer nuclear membrane; PMN, piecemeal microautophagy of the nucleus; PND, programmed nuclear death; PNuD, programmed nuclear destruction; rDNA/rRNA, ribosomal DNA/RNA.

Keywords:  Cargo; degradation; homeostasis; macronucleophagy; membrane; micronucleophagy; mode; nucleophagy; nucleus; vacuole

DOI:  https://doi.org/10.1080/15548627.2021.1971380
Cell Rep. 2021 Oct 12. pii: S2211-1247(21)01260-2. [Epub ahead of print]37(2): 109800

Functional heterogeneity of POMC neurons relies on mTORC1 signaling.

Nicolas Saucisse, Wilfrid Mazier, Vincent Simon, Elke Binder, Caterina Catania, Luigi Bellocchio, Roman A Romanov, Stéphane Léon, Isabelle Matias, Philippe Zizzari, Carmelo Quarta, Astrid Cannich, Kana Meece, Delphine Gonzales, Samantha Clark, Julia M Becker, Giles S H Yeo, Xavier Fioramonti, Florian T Merkle, Sharon L Wardlaw, Tibor Harkany, Federico Massa, Giovanni Marsicano, Daniela Cota.

  Hypothalamic pro-opiomelanocortin (POMC) neurons are known to trigger satiety. However, these neuronal cells encompass heterogeneous subpopulations that release γ-aminobutyric acid (GABA), glutamate, or both neurotransmitters, whose functions are poorly defined. Using conditional mutagenesis and chemogenetics, we show that blockade of the energy sensor mechanistic target of rapamycin complex 1 (mTORC1) in POMC neurons causes hyperphagia by mimicking a cellular negative energy state. This is associated with decreased POMC-derived anorexigenic α-melanocyte-stimulating hormone and recruitment of POMC/GABAergic neurotransmission, which is restrained by cannabinoid type 1 receptor signaling. Electrophysiology and optogenetic studies further reveal that pharmacological blockade of mTORC1 simultaneously activates POMC/GABAergic neurons and inhibits POMC/glutamatergic ones, implying that the functional specificity of these subpopulations relies on mTORC1 activity. Finally, POMC neurons with different neurotransmitter profiles possess specific molecular signatures and spatial distribution. Altogether, these findings suggest that mTORC1 orchestrates the activity of distinct POMC neurons subpopulations to regulate feeding behavior.

Keywords:  CB(1) receptor; Endocannabinoid; Food intake; GABA; Glutamate; Melanocortin; POMC neuron; mTOR

DOI:  https://doi.org/10.1016/j.celrep.2021.109800
Cell Death Discov. 2021 Oct 12. 7(1): 286

The selective degradation of sirtuins via macroautophagy in the MPP+ model of Parkinson's disease is promoted by conserved oxidation sites.

Marius W Baeken, Mario Schwarz, Andreas Kern, Bernd Moosmann, Parvana Hajieva, Christian Behl.

The sirtuin (SIRT) protein family has been of major research interest over the last decades because of their involvement in aging, cancer, and cell death. SIRTs have been implicated in gene and metabolic regulation through their capacity to remove acyl groups from lysine residues in proteins in an NAD+-dependent manner, which may alter individual protein properties as well as the histone-DNA interaction. Since SIRTs regulate a wide range of different signaling cascades, a fine-tuned homeostasis of these proteins is imperative to guarantee the function and survival of the cell. So far, however, how exactly this homeostasis is established has remained unknown. Here, we provide evidence that neuronal SIRT degradation in Parkinson's disease (PD) models is executed by autophagy rather than the proteasome. In neuronal Lund human mesencephalic (LUHMES) cells, all seven SIRTs were substrates for autophagy and showed an accelerated autophagy-dependent degradation upon 1-methyl-4-phenylpyridinium (MPP+) mediated oxidative insults in vitro, whereas the proteasome did not contribute to the removal of oxidized SIRTs. Through blockade of endogenous H2O2 generation and supplementation with the selective radical scavenger phenothiazine (PHT), we could identify H2O2-derived species as the responsible SIRT-oxidizing agents. Analysis of all human SIRTs suggested a conserved regulatory motif based on cysteine oxidation, which may have triggered their degradation via autophagy. High amounts of H2O2, however, rapidly carbonylated selectively SIRT2, SIRT6, and SIRT7, which were found to accumulate carbonylation-prone amino acids. Our data may help in finding new strategies to maintain and modify SIRT bioavailability in neurodegenerative disorders.

DOI: https://doi.org/10.1038/s41420-021-00683-x
Respir Investig. 2021 Oct 11. pii: S2212-5345(21)00158-1. [Epub ahead of print]

Cellular senescence-an aging hallmark in chronic obstructive pulmonary disease pathogenesis.

Jun Araya, Kazuyoshi Kuwano.

  Chronic obstructive pulmonary disease (COPD),1 a representative aging-related pulmonary disorder, is mainly caused by cigarette smoke (CS) exposure. Age is one of the most important risk factors for COPD development, and increased cellular senescence in tissues and organs is a component of aging. CS exposure can induce cellular senescence, as characterized by irreversible growth arrest and aberrant cytokine secretion of the senescence-associated secretory phenotype; thus, accumulation of senescent cells is widely implicated in COPD pathogenesis. CS-induced oxidative modifications to cellular components may be causally linked to accelerated cellular senescence, especially during accumulation of damaged macromolecules. Autophagy is a conserved mechanism whereby cytoplasmic components are sent for lysosomal degradation to maintain proteostasis. Autophagy diminishes with age, and loss of proteostasis is one of the hallmarks of aging. We have reported the involvement of insufficient autophagy in regulating CS-induced cellular senescence with respect to COPD pathogenesis. However, the role of autophagy in COPD pathogenesis can vary based on levels of cell stress and type of selective autophagy because excessive activation of autophagy can be responsible for inducing regulated cell death. Senotherapies targeting cellular senescence may be effective COPD treatments. Autophagy activation could be a promising sonotherapeutic approach, but the optimal modality of autophagy activation should be examined in future studies.

Keywords:  Autophagy; Chronic obstructive pulmonary disease; Lysosome; Senescence; Senotherapy

DOI:  https://doi.org/10.1016/j.resinv.2021.09.003
FEBS Lett. 2021 Oct 15.

Activation of the purinergic receptor P2X7 improves hepatosteatosis by promoting lipophagy.

Zizhi Dong, Yujia Wei, Min Tao, Lili Zhang.

  Nonalcoholic fatty liver disease (NAFLD) is a global health problem that develops through unclear molecular mechanisms. The P2X7 purinergic receptor (P2RX7) is an ATP-gated ion channel that belongs to the P2XR family. Thus far, studies on P2RX7 in NAFLD have been largely contradictory. Integrating experiments and modeling, we elucidate the dynamic processes of lipid droplet fusion and degradation following regulation of P2RX7. We show that activation of P2RX7 can activate the AMPK/ULK1 pathway to promote autophagosome generation and lysosomal degradation of autophagosomes. Inhibiting P2RX7 has the opposite effect. Notably, we find that lipid droplets become larger by the fusion of dysfunctional lysosomes but cannot be degraded by them following P2RX7 inhibition. Our study provides evidence that P2RX7 activation improves NAFLD by promoting lipophagy.

Keywords:  Autophagy; Lipophagy; Nonalcoholic fatty liver disease; Purinergic receptor P2X7

DOI:  https://doi.org/10.1002/1873-3468.14207
Nat Commun. 2021 Oct 13. 12(1): 5991

PGRMC1 acts as a size-selective cargo receptor to drive ER-phagic clearance of mutant prohormones.

Yu-Jie Chen, Jeffrey Knupp, Anoop Arunagiri, Leena Haataja, Peter Arvan, Billy Tsai.

The reticulon-3 (RTN3)-driven targeting complex promotes clearance of misfolded prohormones from the endoplasmic reticulum (ER) for lysosomal destruction by ER-phagy. Because RTN3 resides in the cytosolic leaflet of the ER bilayer, the mechanism of selecting misfolded prohormones as ER-phagy cargo on the luminal side of the ER membrane remains unknown. Here we identify the ER transmembrane protein PGRMC1 as an RTN3-binding partner. Via its luminal domain, PGRMC1 captures misfolded prohormones, targeting them for RTN3-dependent ER-phagy. PGRMC1 selects cargos that are smaller than the large size of other reported ER-phagy substrates. Cargos for PGRMC1 include mutant proinsulins that block secretion of wildtype proinsulin through dominant-negative interactions within the ER, causing insulin-deficiency. Chemical perturbation of PGRMC1 partially restores WT insulin storage by preventing ER-phagic degradation of WT and mutant proinsulin. Thus, PGRMC1 acts as a size-selective cargo receptor during RTN3-dependent ER-phagy, and is a potential therapeutic target for diabetes.

DOI: https://doi.org/10.1038/s41467-021-26225-8
Cell Death Dis. 2021 Oct 13. 12(10): 939

Prefused lysosomes cluster on autophagosomes regulated by VAMP8.

Qixin Chen, Mingang Hao, Lei Wang, Linsen Li, Yang Chen, Xintian Shao, Zhiqi Tian, Richard A Pfuetzner, Qing Zhong, Axel T Brunger, Jun-Lin Guan, Jiajie Diao.

Lysosome-autophagosome fusion is critical to autophagosome maturation. Although several proteins that regulate this fusion process have been identified, the prefusion architecture and its regulation remain unclear. Herein, we show that upon stimulation, multiple lysosomes form clusters around individual autophagosomes, setting the stage for membrane fusion. The soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein on lysosomes-vesicle-associated membrane protein 8 (VAMP8)-plays an important role in forming this prefusion state of lysosomal clusters. To study the potential role of phosphorylation on spontaneous fusion, we investigated the effect of phosphorylation of C-terminal residues of VAMP8. Using a phosphorylation mimic, we observed a decrease of fusion in an ensemble lipid mixing assay and an increase of unfused lysosomes associated with autophagosomes. These results suggest that phosphorylation not only reduces spontaneous fusion for minimizing autophagic flux under normal conditions, but also preassembles multiple lysosomes to increase the fusion probability for resuming autophagy upon stimulation. VAMP8 phosphorylation may thus play an important role in chemotherapy drug resistance by influencing autophagosome maturation.

DOI: https://doi.org/10.1038/s41419-021-04243-0
Cell Death Dis. 2021 Oct 15. 12(10): 950

CHD1L augments autophagy-mediated migration of hepatocellular carcinoma through targeting ZKSCAN3.

Xiaofeng Zhang, Yinshan Bai, Li Huang, Shanshan Liu, Yanxuan Mo, Wei Cheng, Guangliang Wang, Zhiming Cao, Xiaogang Chen, Huiqing Cui, Ling Qi, Lei Ma, Ming Liu, Xin-Yuan Guan, Ning-Fang Ma.

Autophagy is an important biological process in normal cells. However, how it affects tumor progression still remains poorly understood. Herein, we demonstrated that the oncogenic protein Chromodomain-helicase-DNA-binding-protein 1-like gene (CHD1L) might promote HCC cells migration and metastasis through autophagy. CHD1L could bind to the promotor region of Zinc finger with KRAB and SCAN domain 3 (ZKSCAN3), a pivotal autophagy suppressor, and inhibit its transcription. We established inducible CHD1L conditional knockout cell line (CHD1L-iKO cell) and found that the deletion of CHD1L significantly increased ZKSCAN3 expression both at mRNA and protein level. Deletion of CHD1L impaired the autophagic flux and migration of HCC cells, while specifically inhibiting ZKSCAN3 blocked these effects. Further exploration demonstrated that the enhanced tumor cell migration and metastasis induced by CHD1L was mediated through ZKSCAN3-induced autophagic degradation of Paxillin. In summary, we have characterized a previously unknown function of CHD1L in regulating tumor migration via ZKSCAN3-mediated autophagy in HCC. Further inhibition of CHD1L and its downstream autophagy signaling might shed new light on cancer therapeutics.

DOI: https://doi.org/10.1038/s41419-021-04254-x
J Biol Chem. 2021 Oct 08. pii: S0021-9258(21)01096-6. [Epub ahead of print] 101291

Tyrosine Phosphorylation of DEPTOR Functions as a Molecular Switch to Activate mTOR Signaling.

Laurence M Gagné, Nadine Morin, Noémie Lavoie, Nicolas Bisson, Jean-Philippe Lambert, Frédérick A Mallette, Marc-Étienne Huot.

  Metabolic dysfunction is a major driver of tumorigenesis. The serine/threonine kinase mTOR constitutes a key central regulator of metabolic pathways promoting cancer cell proliferation and survival. mTOR activity is regulated by metabolic sensors as well as by numerous factors comprising the PTEN/PI3K/AKT canonical pathway, which are often mutated in cancer. However, some cancers displaying constitutively active mTOR do not carry alterations within this canonical pathway, suggesting alternative modes of mTOR regulation. Since DEPTOR, an endogenous inhibitor of mTOR, was previously found to modulate both mTOR complexe 1 and 2, we investigated the different post-transltionnal modification that could affect its inhibitory function. We found that tyrosine 289 phosphorylation of DEPTOR impairs its interaction with mTOR, leading to increased mTOR activation. Using proximity biotinylation assays, we identified SYK (Spleen tyrosine kinase) as a kinase involved in DEPTOR tyrosine 289 phosphorylation in an ephrin (EPH) receptor-dependent manner. Altogether, our work reveals that phosphorylation of tyrosine 289 of DEPTOR represents a novel molecular switch involved in the regulation of both mTORC1 and mTORC2.

Keywords:  DEPTOR; EPHB2; mTOR; tyrosine phosphorylation

DOI:  https://doi.org/10.1016/j.jbc.2021.101291
Ann Immunol Immunother. 2021 ;pii: 134. [Epub ahead of print]3(1):

How Oncogenic Viruses Exploit p62-Mediated Selective Autophagy for Cancer Development.

Shunbin Ning, Ling Wang.
IUBMB Life. 2021 Oct 15.

Autophagy and cancer metabolism-The two-way interplay.

Reui-Liang Yan, Ruey-Hwa Chen.

  Autophagy is an intracellular catabolic process that degrades cytoplasmic components for recycling in response to stressed conditions, such as nutrient deprivation. Dysregulation of autophagy is associated with various diseases, including cancer. Although autophagy plays dichotomous and context-dependent roles in cancer, evidence has emerged that cancer cells exploit autophagy for metabolic adaptation. Autophagy is upregulated in many cancer types through tumor cell-intrinsic proliferation demands and the hypoxic and nutrient-limited tumor microenvironment (TME). Autophagy-induced breakdown products then fuel into various metabolic pathways to supply tumor cells with energy and building blocks for biosynthesis and survival. This bidirectional regulation between autophagy and tumor constitutes a vicious cycle to potentiate tumor growth and therapy resistance. In addition, the pro-tumor functions of autophagy are expanded to host, including cells in TME and distant organs. Thus, inhibition of autophagy or autophagy-mediated metabolic reprogramming may be a promising strategy for anticancer therapy. Better understanding the metabolic rewiring mechanisms of autophagy for its pro-tumor effects will provide insights into patient treatment.

Keywords:  anticancer therapy; autophagy; cancer; metabolism; tumor microenvironment

DOI:  https://doi.org/10.1002/iub.2569
Front Cell Dev Biol. 2021 ;9 750382

Mitophagy in Diabetic Cardiomyopathy: Roles and Mechanisms.

Haoxiao Zheng, Hailan Zhu, Xinyue Liu, Xiaohui Huang, Anqing Huang, Yuli Huang.

  Cardiovascular disease is the leading complication of diabetes mellitus (DM), and diabetic cardiomyopathy (DCM) is a major cause of mortality in diabetic patients. Multiple pathophysiologic mechanisms, including myocardial insulin resistance, oxidative stress and inflammation, are involved in the development of DCM. Recent studies have shown that mitochondrial dysfunction makes a substantial contribution to the development of DCM. Mitophagy is a type of autophagy that takes place in dysfunctional mitochondria, and it plays a key role in mitochondrial quality control. Although the precise molecular mechanisms of mitophagy in DCM have yet to be fully clarified, recent findings imply that mitophagy improves cardiac function in the diabetic heart. However, excessive mitophagy may exacerbate myocardial damage in patients with DCM. In this review, we aim to provide a comprehensive overview of mitochondrial quality control and the dual roles of mitophagy in DCM. We also propose that a balance between mitochondrial biogenesis and mitophagy is essential for the maintenance of cellular metabolism in the diabetic heart.

Keywords:  diabetic cardiomyopathy; mitochondrial biogenesis; mitochondrial dynamics; mitochondrial quality control; mitophagy

DOI:  https://doi.org/10.3389/fcell.2021.750382
Brain Commun. 2021 ;3(4): fcab222

Epilepsy in the mTORopathies: opportunities for precision medicine.

Patrick B Moloney, Gianpiero L Cavalleri, Norman Delanty.

  The mechanistic target of rapamycin signalling pathway serves as a ubiquitous regulator of cell metabolism, growth, proliferation and survival. The main cellular activity of the mechanistic target of rapamycin cascade funnels through mechanistic target of rapamycin complex 1, which is inhibited by rapamycin, a macrolide compound produced by the bacterium Streptomyces hygroscopicus. Pathogenic variants in genes encoding upstream regulators of mechanistic target of rapamycin complex 1 cause epilepsies and neurodevelopmental disorders. Tuberous sclerosis complex is a multisystem disorder caused by mutations in mechanistic target of rapamycin regulators TSC1 or TSC2, with prominent neurological manifestations including epilepsy, focal cortical dysplasia and neuropsychiatric disorders. Focal cortical dysplasia type II results from somatic brain mutations in mechanistic target of rapamycin pathway activators MTOR, AKT3, PIK3CA and RHEB and is a major cause of drug-resistant epilepsy. DEPDC5, NPRL2 and NPRL3 code for subunits of the GTPase-activating protein (GAP) activity towards Rags 1 complex (GATOR1), the principal amino acid-sensing regulator of mechanistic target of rapamycin complex 1. Germline pathogenic variants in GATOR1 genes cause non-lesional focal epilepsies and epilepsies associated with malformations of cortical development. Collectively, the mTORopathies are characterized by excessive mechanistic target of rapamycin pathway activation and drug-resistant epilepsy. In the first large-scale precision medicine trial in a genetically mediated epilepsy, everolimus (a synthetic analogue of rapamycin) was effective at reducing seizure frequency in people with tuberous sclerosis complex. Rapamycin reduced seizures in rodent models of DEPDC5-related epilepsy and focal cortical dysplasia type II. This review outlines a personalized medicine approach to the management of epilepsies in the mTORopathies. We advocate for early diagnostic sequencing of mechanistic target of rapamycin pathway genes in drug-resistant epilepsy, as identification of a pathogenic variant may point to an occult dysplasia in apparently non-lesional epilepsy or may uncover important prognostic information including, an increased risk of sudden unexpected death in epilepsy in the GATORopathies or favourable epilepsy surgery outcomes in focal cortical dysplasia type II due to somatic brain mutations. Lastly, we discuss the potential therapeutic application of mechanistic target of rapamycin inhibitors for drug-resistant seizures in GATOR1-related epilepsies and focal cortical dysplasia type II.

Keywords:  GATOR1-related epilepsies; everolimus; focal cortical dysplasia type II; the mTORopathies; tuberous sclerosis complex

DOI:  https://doi.org/10.1093/braincomms/fcab222
Biochem Biophys Res Commun. 2021 Oct 09. pii: S0006-291X(21)01399-1. [Epub ahead of print]581 31-37

USP30-AS1 contributes to mitochondrial quality control in glioblastoma cells.

Ningchao Wang, Jiao Li, Qilei Xin, Naihan Xu.

  Glioblastoma is the most serious type of brain cancer with poor prognosis. Here, using the publicly available glioma database, we identified that USP30-AS1, an antisense lncRNA locating on the opposite strand of USP30 locus, is upregulated in human gliomas, particularly in high grade glioma. High level of USP30-AS1 is correlated with poor survival in both primary and recurrent glioma patients. USP30-AS1 regulates mitochondrial homeostasis and mitophagy in glioblastoma cells. Knockdown of USP30-AS1 decreases mitochondrial protein expression and mitochondrial mass, promotes mitochondrial uncoupler-induced mitophagy. However, USP30-AS1 does not regulate USP30 expression in a cis-regulatory manner. In summary, this study proposed that USP30-AS1 may serve as a valuable prognostic marker for gliomas. USP3-AS1 is a negative regulator of mitophagy and the regulatory effect is USP30-independent. USP30-AS1 mediated repression of mitophagy may contribute to the loss of mitochondrial homeostasis and tumor development in glioma.

Keywords:  Glioblastoma; LncRNA; Mitochondria; Mitophagy; USP30-AS1

DOI:  https://doi.org/10.1016/j.bbrc.2021.10.006
Dev Cell. 2021 Oct 11. pii: S1534-5807(21)00735-8. [Epub ahead of print]56(19): 2681-2682

A leucine-derived fatty acid unlocks the mTOR to development.

Chung-Yang Yeh, Dudley W Lamming.

Understanding how nutrient-sensitive signaling pathways regulate development and aging is an active area of research. In this issue of Developmental Cell,Zhu and colleagues (2021) identify a specific monomethylated branched-chain fatty acid that overrides nutrient deprivation signaling and activates mTORC1 in C. elegans and mammalian cells.

DOI: https://doi.org/10.1016/j.devcel.2021.09.017
Cell Mol Gastroenterol Hepatol. 2021 Oct 06. pii: S2352-345X(21)00203-4. [Epub ahead of print]

Defective lipid droplet - lysosome interaction causes fatty liver disease as evidenced by human mutations in TMEM199 and CCDC115.

L E Larsen, M A W van den Boogert, W A Rios-Ocampo, J C Jansen, D Conlon, P L E Chong, J H M Levels, R E Eilers, V V Sachdev, N Zelcer, T Raabe, M He, N J Hand, J P H Drenth, D J Rader, E S G Stroes, D J Lefeber, J W Jonker, A G Holleboom.

BACKGROUND: Recently, novel inborn errors of metabolism were identified due to mutations in V-ATPase assembly factors TMEM199 and CCDC115. Patients are characterized by generalized protein glycosylation defects, hypercholesterolemia and fatty liver disease. Here, we set out to characterize the lipid and fatty liver phenotype in human plasma, cell models and a mouse model.
METHODS AND RESULTS: Patients with TMEM199 and CCDC115 mutations displayed hyperlipidemia, characterized by increased levels of lipoproteins in the very-low density lipoprotein (VLDL) range. HepG2 hepatoma cells, in which the expression of TMEM199 and CCDC115 was silenced, and iPSC-derived hepatocyte-like cells from patients with TMEM199 mutations showed markedly increased secretion of apolipoprotein B (apoB) compared to controls. A mouse model for TMEM199 deficiency with a CRISPR/Cas9-mediated knock-in of the human A7E mutation had marked hepatic steatosis on chow diet. Plasma N-glycans were hypogalactosylated, consistent with the patient phenotype, but no clear plasma lipid abnormalities were observed in the mouse model. In the siTMEM199 and siCCDC115 HepG2 hepatocyte models, increased numbers and size of lipid droplets were observed, including abnormally large lipid droplets, which colocalized with lysosomes. Excessive de novo lipogenesis, failing oxidative capacity, or elevated lipid uptake were not observed. Further investigation of lysosomal function revealed impaired acidification combined with impaired autophagic capacity.
CONCLUSION: Our data suggest that the hypercholesterolemia in TMEM199 and CCDC115 deficiency is due to increased secretion of apoB-containing particles. This may in turn be secondary to the hepatic steatosis observed in these patients as well as in the mouse model. Mechanistically, we observed impaired lysosomal function characterized by reduced acidification, autophagy and increased lysosomal lipid accumulation. These findings could explain the hepatic steatosis seen in patients and highlight the importance of lipophagy in fatty liver disease. As this pathway remains understudied and its regulation largely untargeted, further exploration of this pathway may offer novel strategies for therapeutic interventions to reduce lipotoxicity in fatty liver disease.

DOI: https://doi.org/10.1016/j.jcmgh.2021.09.013
Curr Neuropharmacol. 2021 Oct 05.

Perspective on mTOR-dependent Protection in Status Epilepticus.

Francesca Biagioni, Roberta Celli, Filippo Sean Giorgi, Ferdinando Nicoletti, Francesco Fornai.

   BACKGROUND: The piriform cortex known as area tempestas owns a high propensity to trigger limbic epileptic seizures. Recent studies on human patients indicate that a resection containing the piriform cortex produces a marked improvement in patients suffering from intractable limbic seizures. This calls for looking back pharmacological and anatomical data on area tempestas. Within the piriform cortex status epilepticus can be induced by impairing desensitization of AMPA receptors. The mechanistic target of rapamycin complex1 (mTORC1) is a promising candidate. <P> Objective: The present perspective joins the novel role of the piriform cortex with recent evidence on the modulation of AMPA receptors under the influence of mTORC1. This is based on recent evidence and preliminary data, which lead to formulate an interaction between mTORC1 and AMPA receptors to mitigate the onset of long-lasting, self-sustaining, neurotoxic status epilepticus. <P> Methods: The perspective grounds its method on recent literature along with the actual experimental procedure to elicit status epilepticus from the piriform cortex and the method to administer the mTORC1 inhibitor rapamycin to mitigate seizure expression and brain damage. <P> Results: The available and present perspective converge to show that rapamycin may disrupt the seizure circuitry initiated in the piriform cortex to mitigate seizure duration, severity, and brain damage. <P> Conclusions: The perspective offered by this manuscript provides a novel scenario to understand refractory epilepsy and self-sustaining status epilepticus. This is expected to provide a beneficial outcome in patients suffering from temporal lobe epilepsy.

Keywords:  area tempestas; autophagy; mTOR; piriform cortex; rapamycin; status epilepticus

DOI:  https://doi.org/10.2174/1570159X19666211005152618
Eur J Immunol. 2021 Oct 14.

mTOR-dependent immunometabolism as Achilles' heel of anticancer therapy.

Clarissa Braun, Thomas Weichhart.

  Immune cells are important constituents of the tumor microenvironment and essential in eradicating tumor cells during conventional therapies or novel immunotherapies. The mechanistic target of rapamycin (mTOR) signaling pathway senses the intra- and extracellular nutrient status, growth factor supply and cell stress-related changes to coordinate cellular metabolism and activation dictating effector and memory functions in mainly all hematopoietic immune cells. In addition, the mTOR complex 1 (mTORC1) and mTORC2 are frequently deregulated and become activated in cancer cells to drive cell transformation, survival, neovascularization, and invasion. In this review we provide an overview of the influence of mTOR complexes on immune and cancer cell function and metabolism. We discuss how mTOR inhibitors aiming to target cancer cells will influence immunometabolic cell functions participating either in anti-tumor responses or favoring tumor cell progression in individual immune cells. We suggest immunometabolism as the weak spot of anticancer therapy and propose to evaluate patients according to their predominant immune cell subtype in the cancer tissue. Advances in metabolic drug development that hold promise for more effective treatments in different types of cancer will have to consider their effects on the immune system. This article is protected by copyright. All rights reserved.

Keywords:  Immunometabolism; cancer treatment; immunotherapy; mTORC1; tumor microenvironment

DOI:  https://doi.org/10.1002/eji.202149270
Aging Dis. 2021 Oct;12(7): 1753-1772

Targeting Mitochondrial Network Disorganization is Protective in C. elegans Models of Huntington's Disease.

Emily Machiela, Paige D Rudich, Annika Traa, Ulrich Anglas, Sonja K Soo, Megan M Senchuk, Jeremy M Van Raamsdonk.

  Huntington's disease (HD) is an adult-onset neurodegenerative disease caused by a trinucleotide CAG repeat expansion in the HTT gene. While the pathogenesis of HD is incompletely understood, mitochondrial dysfunction is thought to be a key contributor. In this work, we used C. elegans models to elucidate the role of mitochondrial dynamics in HD. We found that expression of a disease-length polyglutamine tract in body wall muscle, either with or without exon 1 of huntingtin, results in mitochondrial fragmentation and mitochondrial network disorganization. While mitochondria in young HD worms form elongated tubular networks as in wild-type worms, mitochondrial fragmentation occurs with age as expanded polyglutamine protein forms aggregates. To correct the deficit in mitochondrial morphology, we reduced levels of DRP-1, the GTPase responsible for mitochondrial fission. Surprisingly, we found that disrupting drp-1 can have detrimental effects, which are dependent on how much expression is decreased. To avoid potential negative side effects of disrupting drp-1, we examined whether decreasing mitochondrial fragmentation by targeting other genes could be beneficial. Through this approach, we identified multiple genetic targets that rescue movement deficits in worm models of HD. Three of these genetic targets, pgp-3, F25B5.6 and alh-12, increased movement in the HD worm model and restored mitochondrial morphology to wild-type morphology. This work demonstrates that disrupting the mitochondrial fission gene drp-1 can be detrimental in animal models of HD, but that decreasing mitochondrial fragmentation by targeting other genes can be protective. Overall, this study identifies novel therapeutic targets for HD aimed at improving mitochondrial health.

Keywords:  C. elegans; DRP1; Huntington’s disease; aggregation; animal model; genetics; mitochondria; mitochondrial dynamics; neurodegeneration; neuroprotection

DOI:  https://doi.org/10.14336/AD.2021.0404
FEBS Open Bio. 2021 Oct 13.

Metformin may induce ferroptosis by inhibiting autophagy via lncRNA H19 in breast cancer.

Jida Chen, Chuan Qin, Yulu Zhou, Yongxia Chen, Misha Mao, Jingjing Yang.

  Autophagy and ferroptosis have been major foci of biomedical research in recent years. Elucidation of their intrinsic molecular relationships is important for cancer prevention and treatment. Metformin can directly inhibit tumorigenesis, but the mechanism is not fully understood. Here, we demonstrate that metformin and lncRNA-H19 can regulate both autophagy and ferroptosis. Autophagy inducers and H19 can reverse the production of lipid ROS and the inhibition of autophagy induced by metformin. This study suggests that metformin may induce ferroptosis by inhibiting autophagy via H19, and this discovery may facilitate the development of novel therapies for the treatment of breast cancer.

Keywords:  Autophagy; Breast cancer; Ferroptosis; H19; Metformin

DOI:  https://doi.org/10.1002/2211-5463.13314
Behav Brain Res. 2021 Oct 09. pii: S0166-4328(21)00513-1. [Epub ahead of print]417 113625

GluN2A-ERK-mTOR pathway confers a vulnerability to LPS-induced depressive-like behaviour.

Ester Francija, Iva Lukic, Zorica Petrovic, Zeljka Brkic, Milos Mitic, Jelena Radulovic, Miroslav Adzic.

  Inflammation plays a key role in the pathogenesis of the major depressive disorder. Namely, neuroinflammation can induce the production of neuroactive metabolites that interfere with N-methyl-D-aspartate receptors (NMDAR)-mediated glutamatergic neurotransmission and contribute to depressive-like behaviour. On the other hand, mammalian target of rapamycin (mTOR) activity with synaptogenic effects is the main mediator of antidepressant effects of several potent NMDAR antagonists. In this study, we investigated the specific role of GluN2A subunits of NMDAR on the activity of mTOR signaling and behaviour in lipopolysaccharide (LPS)-induces model of depression. The results showed that mice lacking GluN2A subunit did not display depressive-like behavior after the immune challenge, opposite to LPS-treated wild-type mice. Specifically, in GluN2A knockout mice, we estimated the activity of the mTOR pathway in the hippocampus and prefrontal cortex (PFC) by measuring synaptic levels of upstream regulators (p-Akt, p-ERK, and p-GSK3β) and downstream effectors (p-mTOR, and p-p70S6K) of mTOR activity. In addition, we assessed the changes in the levels of two important synaptic markers, GluA1 and PSD-95. Contrary to downregulated mTOR signaling and decreased synaptic markers in LPS-treated wild-type animals, the resilience of GluN2A KO mice to depressive-like behaviour was paralleled with sustained mTOR signaling activity synaptic stability in hippocampus and PFC. Finally, we disclosed that resistance of GluN2A knockouts to LPS-induced depressive-like behavior was ERK-dependent. These findings demonstrate that GluN2A-ERK-mTOR signaling is a vulnerability factor of inflammation-related depressive behaviour, making this signaling pathway the promising target for developing novel antidepressants.

Keywords:  GluN2A knockout mice; Glutamatergic neurotransmission; LPS-induced depression; Synaptosomes; mTOR signaling

DOI:  https://doi.org/10.1016/j.bbr.2021.113625
ACS Chem Neurosci. 2021 Oct 15.

OAB-14 Effectively Ameliorates the Dysfunction of the Endosomal-Autophagic-Lysosomal Pathway in APP/PS1 Transgenic Mice.

Xiaoli Guo, Xuefei Bao, Xiaojuan Wang, Danyang Liu, Peng Liu, Tianyan Chi, Xuefei Ji, Zhonghui Zheng, Guoliang Chen, Libo Zou.

  In Alzheimer's disease (AD), damaged Aβ clearance contributes to elevated levels of Aβ that cause a series of cytotoxic cascade reactions. Thus, targeting Aβ clearance has now been considered a valid therapeutic approach for AD. Cellular uptake and degradation are important mechanisms for Aβ clearance, which are mainly performed by the endosomal-autophagic-lysosomal (EAL) pathway. Our previous study showed that OAB-14, a novel small molecule designed with bexarotene as the lead compound, treatment for 3 months significantly alleviated cognitive disorders and remarkably reduced the deposition of Aβ without affecting its production in APP/PS1 transgenic mice. Here, we further revealed that enhancement of the EAL activity is one of the mechanisms that increases Aβ clearance after OAB-14 administration for 3 months. OAB-14 facilitates receptor-mediated endocytosis and restores autophagy flux via the AMPK/mTOR pathway. Meanwhile, OAB-14 enhances the lysosomal activity, and reduced Aβ accumulation in lysosomes was observed in OAB-14-treated AD mice. These results suggest that OAB-14 may promote Aβ clearance in lysosomes by alleviating the EAL dysfunction in AD mice.

Keywords:  Alzheimer’s disease; OAB-14; amyloid-β; clearance; endosomal-autophagic-lysosomal; memory impairment

DOI:  https://doi.org/10.1021/acschemneuro.1c00209
Front Cell Dev Biol. 2021 ;9 722960

Sestrin2-Mediated Autophagy Contributes to Drug Resistance via Endoplasmic Reticulum Stress in Human Osteosarcoma.

Zhen Tang, Xinghui Wei, Tian Li, Wei Wang, Hao Wu, Hui Dong, Yichao Liu, Feilong Wei, Lei Shi, Xiaokang Li, Zheng Guo, Xin Xiao.

  One contributor to the high mortality of osteosarcoma is its reduced sensitivity to chemotherapy, but the mechanism involved is unclear. Improving the sensitivity of osteosarcoma to chemotherapy is urgently needed to improve patient survival. We found that chemotherapy triggered apoptosis of human osteosarcoma cells in vitro and in vivo; this was accompanied by increased Sestrin2 expression. Importantly, autophagy was also enhanced with increased Sestrin2 expression. Based on this observation, we explored the potential role of Sestrin2 in autophagy of osteosarcoma. We found that Sestrin2 inhibited osteosarcoma cell apoptosis by promoting autophagy via inhibition of endoplasmic reticulum stress, and this process is closely related to the PERK-eIF2α-CHOP pathway. In addition, our study showed that low Sestrin2 expression can effectively reduce autophagy of human osteosarcoma cells after chemotherapy, increase p-mTOR expression, decrease Bcl-2 expression, promote osteosarcoma cell apoptosis, and slow down tumour progression in NU/NU mice. Sestrin2 activates autophagy by inhibiting mTOR via the PERK-eIF2α-CHOP pathway and inhibits apoptosis via Bcl-2. Therefore, our results explain one underlying mechanism of increasing the sensitivity of osteosarcoma to chemotherapy and suggest that Sestrin2 is a promising gene target.

Keywords:  Sestrin2; apoptosis; autophagy; drug resistance; endoplasmic reticulum stress

DOI:  https://doi.org/10.3389/fcell.2021.722960
Stem Cell Reports. 2021 Sep 30. pii: S2213-6711(21)00486-0. [Epub ahead of print]

Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies.

Carolina Ortiz-Cordero, Claudia Bincoletto, Neha R Dhoke, Sridhar Selvaraj, Alessandro Magli, Haowen Zhou, Do-Hyung Kim, Anne G Bang, Rita C R Perlingeiro.

  Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell-derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell-derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.

Keywords:  dystroglycanopathies; iPS cells; in vitro modeling; skeletal muscle

DOI:  https://doi.org/10.1016/j.stemcr.2021.09.009
Elife. 2021 Oct 11. pii: e60478. [Epub ahead of print]10

IFN-γ mediates Paneth cell death via suppression of mTOR.

Alessandra Araujo, Alexandra Safronova, Elise Burger, Américo López-Yglesias, Shilpi Giri, Ellie T Camanzo, Andrew T Martin, Sergei Grivennikov, Felix Yarovinsky.

  Paneth cells constitutively produce antimicrobial peptides and growth factors that allow for intestinal homeostasis, host protection and intestinal stem cell replication. Paneth cells rely heavily on the glycolytic metabolic program, which is in part controlled by the kinase complex Mechanistic target of rapamycin (mTORC1). Yet, little is known about mTOR importance in Paneth cell integrity under steady state and inflammatory conditions. Our results demonstrate that IFN-γ, a crucial mediator of the intestinal inflammation, acts directly on murine Paneth cells to alter their mitochondrial integrity and membrane potential, resulting in an mTORC1-dependent cell death mechanism distinct from canonical cell death pathways including apoptosis, necroptosis, and pyroptosis. These results were established with the purified cytokine and a physiologically relevant common Th1-inducing human parasite Toxoplasma gondii. Given the crucial role for IFN-γ, which is a cytokine frequently associated with the development of inflammatory bowel disease (IBD) and compromised Paneth cell functions, the identified mechanisms underlying mTORC1-dependent Paneth cell death downstream of IFN-γ may provide promising novel approaches for treating intestinal inflammation.

Keywords:  immunology; inflammation; mouse

DOI:  https://doi.org/10.7554/eLife.60478
J Cell Physiol. 2021 Oct 12.

Regulation of electrogenic Na+ /HCO3 - cotransporter 1 (NBCe1) function and its dependence on m-TOR mediated phosphorylation of Ser245.

Marina Giannaki, Christina Ludwig, Stephan Heermann, Eleni Roussa.

  Astrocytes are pivotal responders to alterations of extracellular pH, primarily by regulation of their principal acid-base transporter, the membrane-bound electrogenic Na+ /bicarbonate cotransporter 1 (NBCe1). Here, we describe amammalian target of rapamycin (mTOR)-dependent and NBCe1-mediated astroglial response to extracellular acidosis. Using primary mouse cortical astrocytes, we investigated the effect of long-term extracellular metabolic acidosis on regulation of NBCe1 and elucidated the underlying molecular mechanisms by immunoblotting, biotinylation of surface proteins, intracellular H+ recording using the H+ -sensitive dye 2',7'-bis-(carboxyethyl)-5-(and-6)-carboxyfluorescein, and phosphoproteomic analysis. The results showed significant increase of NBCe1-mediated recovery of intracellular pH from acidification in WT astrocytes, but not in cortical astrocytes from NBCe1-deficient mice. Acidosis-induced upregulation of NBCe1 activity was prevented following inhibition of mTOR signaling by rapamycin. Yet, during acidosis or following exposure of astrocytes to rapamycin, surface protein abundance of NBCe1 remained -unchanged. Mutational analysis in HeLa cells suggested that NBCe1 activity was dependent on phosphorylation state of Ser245 , a residue conserved in all NBCe1 variants. Moreover, phosphorylation state of Ser245 is regulated by mTOR and is inversely correlated with NBCe1 transport activity. Our results identify pSer245 as a novel regulator of NBCe1 functional expression. We propose that context-dependent and mTOR-mediated multisite phosphorylation of serine residues of NBCe1 is likely to be a potent mechanism contributing to the response of astrocytes to acid/base challenges during pathophysiological conditions.

Keywords:  acid-base; acidosis; astrocytes; pH; signaling

DOI:  https://doi.org/10.1002/jcp.30601
Expert Opin Ther Targets. 2021 Oct 12.

Therapeutic potential of PGC-1α in age-related macular degeneration (AMD) - the involvement of mitochondrial quality control, autophagy, and antioxidant response.

Juha M T Hyttinen, Janusz Blasiak, Pasi Tavi, Kai Kaarniranta.

   INTRODUCTION: Age-related macular degeneration (AMD) is the leading, cause of sight loss in the elderly in the Western world. Most patients remain still without any treatment options. The targeting of Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), a transcription co-factor, is a putative therapy against AMD.
AREAS COVERED: The characteristics of AMD and their possible connection with PGC-1α as well as the transcriptional and post-transcriptional control of PGC-1α are discussed. The PGC-1α-driven control of mitochondrial functions, and its involvement in autophagy and antioxidant responses are also examined. Therapeutic possibilities via drugs and epigenetic approaches to enhance PGC-1α expression are discussed. Authors conducted a search of literature mainly from the recent decade from the PubMed database.
EXPERT OPINION: Therapy options in AMD could include PGC-1α activation or stabilization. This could be achieved by a direct elevation of PGC-1α activity, a stabilization or modification of its upstream activators and inhibitors by chemical compounds, like 5-Aminoimidazole-4-carboxamide riboside, metformin, and resveratrol. Furthermore, manipulations with epigenetic modifiers of PGC-1α expression, including miRNAs, e.g. miR-204, are considered. A therapy aimed at PGC-1α up-regulation may be possible in other disorders besides AMD, if they are associated with disturbances in the mitochondria-antioxidant response-autophagy axis.

Keywords:  Age-related macular degeneration; PGC-1α; antioxidant response; autophagy; mitochondria; retinal pigment epithelium

DOI:  https://doi.org/10.1080/14728222.2021.1991913
Int J Mol Sci. 2021 Sep 22. pii: 10208. [Epub ahead of print]22(19):

The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation.

Amalia H Megarioti, Cecilia Primo, George C Kapetanakis, Alexandros Athanasopoulos, Vicky Sophianopoulou, Bruno André, Christos Gournas.

  Selective endocytosis followed by degradation is a major mechanism for downregulating plasma membrane transporters in response to specific environmental cues. In Saccharomyces cerevisiae, this endocytosis is promoted by ubiquitylation catalyzed by the Rsp5 ubiquitin-ligase, targeted to transporters via adaptors of the alpha-arrestin family. However, the molecular mechanisms of this targeting and their control according to conditions remain incompletely understood. In this work, we dissect the molecular mechanisms eliciting the endocytosis of Can1, the arginine permease, in response to cycloheximide-induced TORC1 hyperactivation. We show that cycloheximide promotes Rsp5-dependent Can1 ubiquitylation and endocytosis in a manner dependent on the Bul1/2 alpha-arrestins. Also crucial for this downregulation is a short acidic patch sequence in the N-terminus of Can1 likely acting as a binding site for Bul1/2. The previously reported inhibition by cycloheximide of transporter recycling, from the trans-Golgi network to the plasma membrane, seems to additionally contribute to efficient Can1 downregulation. Our results also indicate that, contrary to the previously described substrate-transport elicited Can1 endocytosis mediated by the Art1 alpha-arrestin, Bul1/2-mediated Can1 ubiquitylation occurs independently of the conformation of the transporter. This study provides further insights into how distinct alpha-arrestins control the ubiquitin-dependent downregulation of a specific amino acid transporter under different conditions.

Keywords:  Nedd4; Npr1; Target of Rapamycin Complex 1; acidic patch; arginine; endocytosis; transporter; ubiquitin; α-arrestin

DOI:  https://doi.org/10.3390/ijms221910208
Cell Rep. 2021 Oct 12. pii: S2211-1247(21)01295-X. [Epub ahead of print]37(2): 109831

Cholecystokinin 1 receptor activation restores normal mTORC1 signaling and is protective to Purkinje cells of SCA mice.

Emily A L Wozniak, Zhao Chen, Sharan Paul, Praseuth Yang, Karla P Figueroa, Jill Friedrich, Tyler Tschumperlin, Michael Berken, Melissa Ingram, Christine Henzler, Stefan M Pulst, Harry T Orr.

  Spinocerebellar ataxias (SCAs) are a group of genetic diseases characterized by progressive ataxia and neurodegeneration, often in cerebellar Purkinje neurons. A SCA1 mouse model, Pcp2-ATXN1[30Q]D776, has severe ataxia in absence of progressive Purkinje neuron degeneration and death. Previous RNA-seq analyses identify cerebellar upregulation of the peptide hormone cholecystokinin (Cck) in Pcp2-ATXN1[30Q]D776 mice. Importantly, absence of Cck1 receptor (Cck1R) in Pcp2-ATXN1[30Q]D776 mice confers a progressive disease with Purkinje neuron death. Administration of a Cck1R agonist, A71623, to Pcp2-ATXN1[30Q]D776;Cck-/- and Pcp2-AXTN1[82Q] mice dampens Purkinje neuron pathology and associated deficits in motor performance. In addition, A71623 administration improves motor performance of Pcp2-ATXN2[127Q] SCA2 mice. Moreover, the Cck1R agonist A71623 corrects mTORC1 signaling and improves expression of calbindin in cerebella of AXTN1[82Q] and ATXN2[127Q] mice. These results indicate that manipulation of the Cck-Cck1R pathway is a potential therapeutic target for treatment of diseases involving Purkinje neuron degeneration.

Keywords:  Purkinje cells; cholecystokinin; mTORC1 signaling; neuroprotection; spinocerebellar ataxia

DOI:  https://doi.org/10.1016/j.celrep.2021.109831
J Dermatol. 2021 Oct 16.

Autophagy in skin barrier and immune-related skin diseases.

Chi Liu, Lei Gu, Jie Ding, Qianchao Meng, Nan Li, Guifeng Dai, Qinying Li, Xueyong Wu.

  Autophagy is a process which is highly conserved in eukaryotes to degrade or recycle cytoplasmic components through lysosomes to maintain cellular homeostasis. Recent studies have shown that autophagy also plays critical roles in cell apoptosis, inflammation, pathogen clearance, and so on under stressed conditions and thereby has been linked to a variety of human disorders. The skin is the largest organ of the body and serves as the first line of defense against environmental insult. Skin as a nutrient-poor environment requires recycling of limited resources via the autophagy machinery to maintain homeostasis. Therefore, dysregulation of autophagy has been linked to skin diseases. In this review, we describe the molecular machinery and regulation of autophagy, discuss its role in keratinocytes and skin barrier, skin immune cells, and immune-related skin diseases including autoimmune skin disorders, allergic skin diseases, infectious skin disorders, and antitumor immunity against skin tumor. Finally, we highlight the potential of autophagy as a therapeutic target for immune-related skin diseases, and delivery of autophagy-related molecules (such as inducers, inhibitors, or nucleic acid molecules) by virtue of physical materials (such as nanoparticles) or biological materials (such as peptides) to skin topically may obtain clinical benefits in immune-related skin diseases. Moreover, developing autophagy-related gene product-based biomarkers may be promising to diagnose immune-related skin diseases.

Keywords:  autophagy; immune cells; immune-related skin diseases; keratinocyte; skin barrier

DOI:  https://doi.org/10.1111/1346-8138.16185