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Histone lactylation stimulated upregulation of PSMD14 alleviates neuron PANoptosis through deubiquitinating PKM2 to activate PINK1-mediated mitophagy after traumatic brain injury.
Pub Date : 2025-03-05 DOI: 10.1080/15548627.2025.2471633
Lei Xu, Yangfan Ye, Wei Gu, Xiao Xu, Nuo Chen, Liuchao Zhang, Wanzhi Cai, Jingming Hu, Tian Wang, Honglu Chao, Yiming Tu, Jing Ji

Alleviating the multiple types of programmed neuronal death caused by mechanical injury has been an impetus for designing neuro-therapeutical approaches after traumatic brain injury (TBI). The aim of this study was to elucidate the potential role of PSMD14 (proteasome 26S subunit, non-ATPase 14) in neuron death and the specific mechanism through which it improves prognosis of TBI patients. Here, we identified differential expression of the PSMD14 protein between the controlled cortical impact (CCI) and sham mouse groups by LC-MS proteomic analysis and found that PSMD14 was significantly upregulated in neurons after brain injury by qPCR and western blot. PSMD14 suppressed stretch-induced neuron PANoptosis and improved motor ability and learning performance after CCI in vivo. Mechanistically, PSMD14 improved PINK1 phosphorylation levels at Thr257 and activated PINK1-mediated mitophagy by deubiquitinating PKM/PKM2 (pyruvate kinase M1/2) to maintain PKM protein stability. PSMD14-induced mitophagy promoted mitochondrial homeostasis to reduced ROS production, and ultimately inhibited the neuron PANoptosis. The upregulation of neuronal PSMD14 after TBI was due to the increase of histone lactation modification level and lactate treatment alleviated neuron PANoptosis via increasing PSMD14 expression. Our findings suggest that PSMD14 could be a potential therapeutic approach for improving the prognosis of TBI patients.Abbreviations: CCI: controlled cortical impact; CQ: chloroquine; DUBs: deubiquitinating enzymes; H3K18la: H3 lysine 18 lactylation; IB: immunoblot; IHC: immunohistochemistry; IP: immunoprecipitation; MLKL: mixed lineage kinase domain like pseudokinase; PI3K: phosphoinositide 3-kinase; PINK1: PTEN induced kinase 1; PKM/PKM2: pyruvate kinase M1/2; PSMD14: proteasome 26S subunit, non-ATPase 14; ROS: reactive oxygen species; RIPK1: receptor interacting serine/threonine kinase 1; RIPK3: receptor interacting serine/threonine kinase 3; TBI: traumatic brain injury.

{"title":"Histone lactylation stimulated upregulation of PSMD14 alleviates neuron PANoptosis through deubiquitinating PKM2 to activate PINK1-mediated mitophagy after traumatic brain injury.","authors":"Lei Xu, Yangfan Ye, Wei Gu, Xiao Xu, Nuo Chen, Liuchao Zhang, Wanzhi Cai, Jingming Hu, Tian Wang, Honglu Chao, Yiming Tu, Jing Ji","doi":"10.1080/15548627.2025.2471633","DOIUrl":"10.1080/15548627.2025.2471633","url":null,"abstract":"<p><p>Alleviating the multiple types of programmed neuronal death caused by mechanical injury has been an impetus for designing neuro-therapeutical approaches after traumatic brain injury (TBI). The aim of this study was to elucidate the potential role of PSMD14 (proteasome 26S subunit, non-ATPase 14) in neuron death and the specific mechanism through which it improves prognosis of TBI patients. Here, we identified differential expression of the PSMD14 protein between the controlled cortical impact (CCI) and sham mouse groups by LC-MS proteomic analysis and found that PSMD14 was significantly upregulated in neurons after brain injury by qPCR and western blot. PSMD14 suppressed stretch-induced neuron PANoptosis and improved motor ability and learning performance after CCI in vivo. Mechanistically, PSMD14 improved PINK1 phosphorylation levels at Thr257 and activated PINK1-mediated mitophagy by deubiquitinating PKM/PKM2 (pyruvate kinase M1/2) to maintain PKM protein stability. PSMD14-induced mitophagy promoted mitochondrial homeostasis to reduced ROS production, and ultimately inhibited the neuron PANoptosis. The upregulation of neuronal PSMD14 after TBI was due to the increase of histone lactation modification level and lactate treatment alleviated neuron PANoptosis via increasing PSMD14 expression. Our findings suggest that PSMD14 could be a potential therapeutic approach for improving the prognosis of TBI patients.<b>Abbreviations:</b> CCI: controlled cortical impact; CQ: chloroquine; DUBs: deubiquitinating enzymes; H3K18la: H3 lysine 18 lactylation; IB: immunoblot; IHC: immunohistochemistry; IP: immunoprecipitation; MLKL: mixed lineage kinase domain like pseudokinase; PI3K: phosphoinositide 3-kinase; PINK1: PTEN induced kinase 1; PKM/PKM2: pyruvate kinase M1/2; PSMD14: proteasome 26S subunit, non-ATPase 14; ROS: reactive oxygen species; RIPK1: receptor interacting serine/threonine kinase 1; RIPK3: receptor interacting serine/threonine kinase 3; TBI: traumatic brain injury.</p>","PeriodicalId":93893,"journal":{"name":"Autophagy","volume":" ","pages":"1-19"},"PeriodicalIF":0.0,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Autophagy regulates cellular senescence by mediating the degradation of CDKN1A/p21 and CDKN2A/p16 through SQSTM1/p62-mediated selective autophagy in myxomatous mitral valve degeneration.
Pub Date : 2025-03-04 DOI: 10.1080/15548627.2025.2469315
Qiyu Tang, Keyi Tang, Greg R Markby, Maciej Parys, Kanchan Phadwal, Vicky E MacRae, Brendan M Corcoran
<p><p>Myxomatous mitral valve degeneration (MMVD) is one of the most important age-dependent degenerative heart valve disorders in both humans and dogs. It is characterized by the aberrant remodeling of extracellular matrix (ECM), regulated by senescent myofibroblasts (aVICs) transitioning from quiescent valve interstitial cells (qVICs), primarily under TGFB1/TGF-β1 control. In the present study, we found senescent aVICs exhibited impaired macroautophagy/autophagy as evidenced by compromised autophagy flux and immature autophagosomes. MTOR-dependent autophagy induced by rapamycin and torin-1 attenuated cell senescence and decreased the expression of cyclin-dependent kinase inhibitors (CDKIs) CDKN2A/p16<sup>INK4A</sup> and CDKN1A/p21<sup>CIP1</sup>. Furthermore, induction of autophagy in aVICs by <i>ATG</i> (autophagy related) gene overexpression restored autophagy flux, with a concomitant reduction in CDKN1A and CDKN2A expression and senescence-associated secretory phenotype (SASP). Conversely, autophagy deficiency induced CDKN1A and CDKN2A accumulation and SASP, whereas ATG re-expression alleviated senescent phenotypic transformation. Notably, CDKN1A and CDKN2A localized to autophagosomes and lysosomes following MTOR antagonism or MG132 treatment. SQSTM1/p62 was identified as the autophagy receptor to selectively sequester CDKN1A and CDKN2A cargoes for autophagic degradation. Our findings are the first demonstration that CDKN1A and CDKN2A are degraded through SQSTM1-mediated selective autophagy, independent of the ubiquitin-proteasome pathway. These data will inform development of therapeutic strategies for the treatment of canine and human MMVD, and for the treatment of Alzheimer disease, Parkinson disease and other age-related degenerative disorders.<b>Abbreviations</b>: ACTA2/α-SMA: actin alpha 2, smooth muscle; AKT: AKT serine/threonine kinase; aVICs: activated valve interstitial cells; ATG: autophagy related; baf-A1: bafilomycin A<sub>1</sub>; BrdU, bromodeoxyuridine; BSA: bovine serum albumin; CDKIs, cyclin-dependent kinase inhibitors; CDKN1A/p21: cyclin dependent kinase inhibitor 1A; CDKN2A/p16: cyclin dependent kinase inhibitor 2A; co-IP: co-immunoprecipitation; DMSO: dimethylsulfoxide; ECM, extracellular matrix; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; eGFP: green fluorescent protein; ELISA: enzyme-linked immunosorbent assay; HEK-293T, human embryonic kidney 293T; HRP: horseradish peroxidase; KO: knockout; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; LIR: MAP1LC3/LC3-interacting region; MFS: Marfan syndrome; MKI67/Ki-67: marker of proliferation Ki-67; MMVD: myxomatous mitral valve degeneration; MTOR: mechanistic target of rapamycin kinase; MTORC: MTOR complex; OE: overexpression; PBST, phosphate-buffered saline with 0.1% Tween-20; PCNA: proliferating cell nuclear antigen; PIK3CA/PI3K: phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; PLA: proximity ligation assays; PS
肌瘤性二尖瓣变性(MMVD)是人类和狗中最重要的年龄依赖性心脏瓣膜退行性疾病之一。它的特征是细胞外基质(ECM)的异常重塑,由衰老的肌成纤维细胞(aVICs)从静止的瓣膜间质细胞(qVICs)转变而来,主要受 TGFB1/TGF-β1 的控制。在本研究中,我们发现衰老的瓣膜间充质细胞(aVICs)的大自噬/自噬功能受损,表现为自噬通量受损和自噬体不成熟。雷帕霉素和 torin-1 诱导的 MTOR 依赖性自噬减轻了细胞衰老,并降低了细胞周期蛋白依赖性激酶抑制剂(CDKIs)CDKN2A/p16INK4A 和 CDKN1A/p21CIP1 的表达。此外,通过过表达 ATG(自噬相关)基因诱导 aVIC 中的自噬,可恢复自噬通量,同时减少 CDKN1A 和 CDKN2A 的表达以及衰老相关分泌表型(SASP)。相反,自噬缺乏会诱导 CDKN1A 和 CDKN2A 的积累和 SASP,而 ATG 的再次表达则会缓解衰老表型的转变。值得注意的是,在MTOR拮抗或MG132处理后,CDKN1A和CDKN2A定位于自噬体和溶酶体。SQSTM1/p62 被鉴定为自噬受体,可选择性地扣留 CDKN1A 和 CDKN2A 货物进行自噬降解。我们的研究结果首次证明,CDKN1A 和 CDKN2A 是通过 SQSTM1 介导的选择性自噬降解的,与泛素-蛋白酶体途径无关。这些数据将为开发治疗犬和人类MMVD以及治疗阿尔茨海默病、帕金森病和其他与年龄相关的退行性疾病的治疗策略提供依据。
{"title":"Autophagy regulates cellular senescence by mediating the degradation of CDKN1A/p21 and CDKN2A/p16 through SQSTM1/p62-mediated selective autophagy in myxomatous mitral valve degeneration.","authors":"Qiyu Tang, Keyi Tang, Greg R Markby, Maciej Parys, Kanchan Phadwal, Vicky E MacRae, Brendan M Corcoran","doi":"10.1080/15548627.2025.2469315","DOIUrl":"10.1080/15548627.2025.2469315","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Myxomatous mitral valve degeneration (MMVD) is one of the most important age-dependent degenerative heart valve disorders in both humans and dogs. It is characterized by the aberrant remodeling of extracellular matrix (ECM), regulated by senescent myofibroblasts (aVICs) transitioning from quiescent valve interstitial cells (qVICs), primarily under TGFB1/TGF-β1 control. In the present study, we found senescent aVICs exhibited impaired macroautophagy/autophagy as evidenced by compromised autophagy flux and immature autophagosomes. MTOR-dependent autophagy induced by rapamycin and torin-1 attenuated cell senescence and decreased the expression of cyclin-dependent kinase inhibitors (CDKIs) CDKN2A/p16&lt;sup&gt;INK4A&lt;/sup&gt; and CDKN1A/p21&lt;sup&gt;CIP1&lt;/sup&gt;. Furthermore, induction of autophagy in aVICs by &lt;i&gt;ATG&lt;/i&gt; (autophagy related) gene overexpression restored autophagy flux, with a concomitant reduction in CDKN1A and CDKN2A expression and senescence-associated secretory phenotype (SASP). Conversely, autophagy deficiency induced CDKN1A and CDKN2A accumulation and SASP, whereas ATG re-expression alleviated senescent phenotypic transformation. Notably, CDKN1A and CDKN2A localized to autophagosomes and lysosomes following MTOR antagonism or MG132 treatment. SQSTM1/p62 was identified as the autophagy receptor to selectively sequester CDKN1A and CDKN2A cargoes for autophagic degradation. Our findings are the first demonstration that CDKN1A and CDKN2A are degraded through SQSTM1-mediated selective autophagy, independent of the ubiquitin-proteasome pathway. These data will inform development of therapeutic strategies for the treatment of canine and human MMVD, and for the treatment of Alzheimer disease, Parkinson disease and other age-related degenerative disorders.&lt;b&gt;Abbreviations&lt;/b&gt;: ACTA2/α-SMA: actin alpha 2, smooth muscle; AKT: AKT serine/threonine kinase; aVICs: activated valve interstitial cells; ATG: autophagy related; baf-A1: bafilomycin A&lt;sub&gt;1&lt;/sub&gt;; BrdU, bromodeoxyuridine; BSA: bovine serum albumin; CDKIs, cyclin-dependent kinase inhibitors; CDKN1A/p21: cyclin dependent kinase inhibitor 1A; CDKN2A/p16: cyclin dependent kinase inhibitor 2A; co-IP: co-immunoprecipitation; DMSO: dimethylsulfoxide; ECM, extracellular matrix; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; eGFP: green fluorescent protein; ELISA: enzyme-linked immunosorbent assay; HEK-293T, human embryonic kidney 293T; HRP: horseradish peroxidase; KO: knockout; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; LIR: MAP1LC3/LC3-interacting region; MFS: Marfan syndrome; MKI67/Ki-67: marker of proliferation Ki-67; MMVD: myxomatous mitral valve degeneration; MTOR: mechanistic target of rapamycin kinase; MTORC: MTOR complex; OE: overexpression; PBST, phosphate-buffered saline with 0.1% Tween-20; PCNA: proliferating cell nuclear antigen; PIK3CA/PI3K: phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; PLA: proximity ligation assays; PS","PeriodicalId":93893,"journal":{"name":"Autophagy","volume":" ","pages":"1-23"},"PeriodicalIF":0.0,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Establishment of a yeast essential protein conditional-degradation library and screening for autophagy-regulating genes.
Pub Date : 2025-03-04 DOI: 10.1080/15548627.2025.2469189
Yi Zhang, Yingcong Chen, Choufei Wu, Zhengyi Cai, Weijing Yao, Huan Yang, Juan Song, Xiankuan Xie, Liqin Zhang, Cong Yi

Macroautophagy/autophagy is an evolutionarily conserved intracellular degradation pathway that relies on vacuoles or lysosomes. Over 40 ATG genes have been identified in yeast cells as participants in various types of autophagy, although these genes are non-essential. While some essential genes involved in autophagy have been identified using temperature-sensitive yeast strains, systematic research on essential genes in autophagy remains lacking. To address this, we established an essential protein conditional degradation library using the auxin-inducible degron (AID) system. By introducing the GFP-Atg8 plasmid, we identified 29 essential yeast genes involved in autophagy, 19 of which had not been previously recognized. In summary, the yeast essential protein conditional degradation library we constructed will serve as a valuable resource for systematically investigating the roles of essential genes in autophagy and other biological functions.Abbreviation: AID: auxin-inducible degron; ALP: alkaline phosphatase; ATG: autophagy related; CSG: constitutive slow growth; DAmP: Decreased Abundance by mRNA Perturbation; GFP: green fluorescent protein; MMS: methyl methanesulfonate; ORF: open reading frame; PAS: phagophore assembly site; PCR: polymerase chain reaction; SD-G: glucose starvation medium; SD-N: nitrogen starvation medium; TOR: target of rapamycin kinase; YGRC: yeast genetic resource center; YPD: yeast extract peptone dextrose.

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引用次数: 0
Mycobacterium bovis Mb3523c protein regulates host ferroptosis via chaperone-mediated autophagy.
Pub Date : 2025-03-03 DOI: 10.1080/15548627.2025.2468139
Haoran Wang, Dingpu Liu, Xin Ge, Yuanzhi Wang, Xiangmei Zhou

The occurrence of necrosis during Mycobacterium bovis (M. bovis) infection is regarded as harmful to the host because it promotes the spread of M. bovis. Ferroptosis is a controlled type of cell death that occurs when there is an excessive buildup of both free iron and harmful lipid peroxides. Here, we demonstrate that the mammalian cell entry (Mce) 4 family protein Mb3523c triggers ferroptosis to promote M. bovis pathogenicity and dissemination. Mechanistically, Mb3523c, through its Y237 and G241 site, interacts with host HSP90 protein to stabilize the LAMP2A on the lysosome to promote the chaperone-mediated autophagy (CMA) pathway. Then, GPX4 is delivered to lysosomes for destruction via the CMA pathway, eventually inducing ferroptosis to promote M. bovis transmission. In summary, our findings offer novel insights into the molecular mechanisms of pathogen-induced ferroptosis, demonstrating that targeting the GPX4-dependent ferroptosis through blocking the M. bovis Mb3523c-host HSP90 interface represents a potential therapeutic strategy for tuberculosis (TB).Abbreviations: CFU: colony-forming units; CMA: chaperone-mediated autophagy; Co-IP: co-immunoprecipitation; Fer-1: ferrostatin-1; GPX4: glutathione peroxidase 4; HSP90: heat shock protein 90; LDH: lactate dehydrogenase; Mce: mammalian cell entry; MOI: multiplicity of infection; Nec-1: necrostatin-1; PI: propidium iodide; RCD: regulated cell death.

{"title":"<i>Mycobacterium bovis</i> Mb3523c protein regulates host ferroptosis via chaperone-mediated autophagy.","authors":"Haoran Wang, Dingpu Liu, Xin Ge, Yuanzhi Wang, Xiangmei Zhou","doi":"10.1080/15548627.2025.2468139","DOIUrl":"10.1080/15548627.2025.2468139","url":null,"abstract":"<p><p>The occurrence of necrosis during <i>Mycobacterium bovis</i> (<i>M. bovis</i>) infection is regarded as harmful to the host because it promotes the spread of <i>M. bovis</i>. Ferroptosis is a controlled type of cell death that occurs when there is an excessive buildup of both free iron and harmful lipid peroxides. Here, we demonstrate that the mammalian cell entry (Mce) 4 family protein Mb3523c triggers ferroptosis to promote <i>M. bovis</i> pathogenicity and dissemination. Mechanistically, Mb3523c, through its Y237 and G241 site, interacts with host HSP90 protein to stabilize the LAMP2A on the lysosome to promote the chaperone-mediated autophagy (CMA) pathway. Then, GPX4 is delivered to lysosomes for destruction via the CMA pathway, eventually inducing ferroptosis to promote <i>M. bovis</i> transmission. In summary, our findings offer novel insights into the molecular mechanisms of pathogen-induced ferroptosis, demonstrating that targeting the GPX4-dependent ferroptosis through blocking the <i>M. bovis</i> Mb3523c-host HSP90 interface represents a potential therapeutic strategy for tuberculosis (TB).<b>Abbreviations</b>: CFU: colony-forming units; CMA: chaperone-mediated autophagy; Co-IP: co-immunoprecipitation; Fer-1: ferrostatin-1; GPX4: glutathione peroxidase 4; HSP90: heat shock protein 90; LDH: lactate dehydrogenase; Mce: mammalian cell entry; MOI: multiplicity of infection; Nec-1: necrostatin-1; PI: propidium iodide; RCD: regulated cell death.</p>","PeriodicalId":93893,"journal":{"name":"Autophagy","volume":" ","pages":"1-18"},"PeriodicalIF":0.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143451313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
TAX1BP1-dependent autophagic degradation of STING1 impairs anti-tumor immunity.
Pub Date : 2025-03-03 DOI: 10.1080/15548627.2025.2471736
Ruoxi Zhang, Chunhua Yu, Herbert J Zeh, Guido Kroemer, Daniel J Klionsky, Daolin Tang, Rui Kang

The activation of STING1 can lead to the production and secretion of cytokines, initiating antitumor immunity. Here, we screened an ion channel ligand library and identified tetrandrine, a bis-benzylisoquinoline alkaloid, as an immunological adjuvant that enhances antitumor immunity by preventing the autophagic degradation of the STING1 protein. This tetrandrine effect is independent of its known function as a calcium or potassium channel blocker. Instead, tetrandrine inhibits lysosomal function, impairing cathepsin maturation, and autophagic degradation. Proteomic analysis of lysosomes identified TAX1BP1 as a novel autophagic receptor for the proteolysis of STING1. TAX1BP1 recognizes STING1 through the physical interaction of its coiled-coil domain with the cyclic dinucleotide binding domain of STING1. Systematic mutation of lysine (K) residues revealed that K63-ubiquitination of STING1 at the K224 site ignites TAX1BP1-dependent STING1 degradation. Combined treatment with tetrandrine and STING1 agonists promotes antitumor immunity by converting "cold" pancreatic cancers into "hot" tumors. This process is associated with enhanced cytokine release and increased infiltration of cytotoxic T-cells into the tumor microenvironment. The antitumor immunity mediated by tetrandrine and STING1 agonists is limited by neutralizing antibodies to the type I interferon receptor or CD8+ T cells. Thus, these findings establish a potential immunotherapeutic strategy against pancreatic cancer by preventing the autophagic degradation of STING1.

{"title":"TAX1BP1-dependent autophagic degradation of STING1 impairs anti-tumor immunity.","authors":"Ruoxi Zhang, Chunhua Yu, Herbert J Zeh, Guido Kroemer, Daniel J Klionsky, Daolin Tang, Rui Kang","doi":"10.1080/15548627.2025.2471736","DOIUrl":"10.1080/15548627.2025.2471736","url":null,"abstract":"<p><p>The activation of STING1 can lead to the production and secretion of cytokines, initiating antitumor immunity. Here, we screened an ion channel ligand library and identified tetrandrine, a bis-benzylisoquinoline alkaloid, as an immunological adjuvant that enhances antitumor immunity by preventing the autophagic degradation of the STING1 protein. This tetrandrine effect is independent of its known function as a calcium or potassium channel blocker. Instead, tetrandrine inhibits lysosomal function, impairing cathepsin maturation, and autophagic degradation. Proteomic analysis of lysosomes identified TAX1BP1 as a novel autophagic receptor for the proteolysis of STING1. TAX1BP1 recognizes STING1 through the physical interaction of its coiled-coil domain with the cyclic dinucleotide binding domain of STING1. Systematic mutation of lysine (K) residues revealed that K63-ubiquitination of STING1 at the K224 site ignites TAX1BP1-dependent STING1 degradation. Combined treatment with tetrandrine and STING1 agonists promotes antitumor immunity by converting \"cold\" pancreatic cancers into \"hot\" tumors. This process is associated with enhanced cytokine release and increased infiltration of cytotoxic T-cells into the tumor microenvironment. The antitumor immunity mediated by tetrandrine and STING1 agonists is limited by neutralizing antibodies to the type I interferon receptor or CD8<sup>+</sup> T cells. Thus, these findings establish a potential immunotherapeutic strategy against pancreatic cancer by preventing the autophagic degradation of STING1.</p>","PeriodicalId":93893,"journal":{"name":"Autophagy","volume":" ","pages":"1-22"},"PeriodicalIF":0.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A molecular glue for PRKN/parkin. 用于Prkn/parkin的分子胶。
Pub Date : 2025-03-01 Epub Date: 2024-12-24 DOI: 10.1080/15548627.2024.2443232
Véronique Sauvé, Kalle Gehring

Parkinson disease (PD) is a neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra, primarily due to mitochondria dysfunction. PRKN (parkin RBR E3 ubiquitin protein ligase) and PINK1 (PTEN induced kinase 1) are linked to early-onset cases of PD and essential for the clearance of damaged mitochondria via selective mitochondrial autophagy (mitophagy). In a recent publication, we detail how a small molecule can activate PRKN mutants that are unable to be phosphorylated, restoring mitophagy in cellular assays. These findings offer hope for the design of therapeutic drugs for some forms of PD.

帕金森病(PD)是一种神经退行性疾病,其特征是黑质多巴胺能神经元的丧失,主要是由于线粒体功能障碍。PRKN (parkin RBR E3泛素蛋白连接酶)和PINK1 (PTEN诱导的激酶1)与早发性PD病例有关,并且通过选择性线粒体自噬(mitophagy)清除受损线粒体至关重要。在最近发表的一篇文章中,我们详细介绍了一个小分子如何激活无法磷酸化的PRKN突变体,在细胞分析中恢复有丝分裂。这些发现为设计治疗某些形式帕金森病的药物提供了希望。
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引用次数: 0
Cargo hitchhiking autophagy - a hybrid autophagy pathway utilized in yeast. 搭便车自噬——酵母中利用的一种杂交自噬途径。
Pub Date : 2025-03-01 Epub Date: 2025-01-05 DOI: 10.1080/15548627.2024.2447207
Katrina F Cooper

Macroautophagy is a catabolic process that maintains cellular homeostasis by recycling intracellular material through the use of double-membrane vesicles called autophagosomes. In turn, autophagosomes fuse with vacuoles (in yeast and plants) or lysosomes (in metazoans), where resident hydrolases degrade the cargo. Given the conservation of autophagy, Saccharomyces cerevisiae is a valuable model organism for deciphering molecular details that define macroautophagy pathways. In yeast, macroautophagic pathways fall into two subclasses: selective and nonselective (bulk) autophagy. Bulk autophagy is predominantly upregulated following TORC1 inhibition, triggered by nutrient stress, and degrades superfluous random cytosolic proteins and organelles. In contrast, selective autophagy pathways maintain cellular homeostasis when TORC1 is active by degrading damaged organelles and dysfunctional proteins. Here, selective autophagy receptors mediate cargo delivery to the vacuole. Now, two groups have discovered a new hybrid autophagy mechanism, coined cargo hitchhiking autophagy (CHA), that uses autophagic receptor proteins to deliver selected cargo to phagophores built in response to nutrient stress for the random destruction of cytosolic contents. In CHA, various autophagic receptors link their cargos to lipidated Atg8, located on growing phagophores. In addition, the sorting nexin heterodimer Snx4-Atg20 assists in the degradation of cargo during CHA, possibly by aiding the delivery of cytoplasmic cargos to phagophores and/or by delaying the closure of expanding phagophores. This review will outline this new mechanism, also known as Snx4-assisted autophagy, that degrades an assortment of cargos in yeast, including transcription factors, glycogen, and a subset of ribosomal proteins.

巨噬是一种分解代谢过程,通过使用称为自噬体的双膜囊泡循环细胞内物质来维持细胞稳态。反过来,自噬体与液泡(在酵母和植物中)或溶酶体(在后生动物中)融合,其中驻留的水解酶降解货物。鉴于自噬的保守性,酿酒酵母是破译定义巨噬途径的分子细节的有价值的模式生物。在酵母中,巨噬途径分为两个亚类:选择性和非选择性(大量)自噬。大量自噬主要在TORC1抑制后上调,由营养胁迫触发,并降解多余的随机细胞质蛋白和细胞器。相反,当TORC1通过降解受损细胞器和功能失调蛋白而激活时,选择性自噬途径维持细胞稳态。在这里,选择性自噬受体介导货物输送到液泡。现在,两个研究小组发现了一种新的杂交自噬机制,称为“搭便车自噬”(CHA),它利用自噬受体蛋白将选定的货物运送到为应对营养胁迫而建立的吞噬团中,以随机破坏细胞质内容物。在CHA中,各种自噬受体将其货物连接到位于生长的吞噬细胞上的脂化at8。此外,分选连接蛋白异二聚体snx4 - at20在CHA过程中有助于货物的降解,可能是通过帮助细胞质货物向吞噬细胞的传递和/或通过延迟扩大的吞噬细胞的关闭。这篇综述将概述这种新机制,也被称为snx4辅助自噬,它降解酵母中的各种货物,包括转录因子、糖原和核糖体蛋白子集。
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引用次数: 0
Autophagy-dependent hepatocyte secretion of DBI/ACBP induced by glucocorticoids determines the pathogenesis of Cushing syndrome. 糖皮质激素诱导的自噬依赖性肝细胞分泌DBI/ACBP决定了库欣综合征的发病机制。
Pub Date : 2025-03-01 Epub Date: 2024-12-30 DOI: 10.1080/15548627.2024.2437649
Hui Pan, Ai-Ling Tian, Fréderic Castinetti, Isabelle Martins, Oliver Kepp, Guido Kroemer

DBI/ACBP is a phylogenetically ancient hormone that stimulates appetite and lipo-anabolism. In response to starvation, DBI/ACBP is secreted through a noncanonical, macroautophagy/autophagy-dependent pathway. The physiological hunger reflex involves starvation-induced secretion of DBI/ACBP from multiple cell types. DBI/ACBP concentrations subsequently increase in extracellular fluids to stimulate food intake. Recently, we observed that glucocorticoids, which are endogenous stress hormones as well as anti-inflammatory drugs, upregulate DBI/ACBP expression at the transcriptional level and stimulate autophagy in hepatocytes, thereby causing a surge in circulating DBI/ACBP levels. Prolonged increase in glucocorticoid concentrations causes an extreme form of metabolic syndrome, dubbed "Cushing syndrome", which is characterized by clinical features including hyperphagia, hyperdipsia, dyslipidemia, hyperinsulinemia, insulin resistance, lipodystrophy, visceral adiposity, steatosis, sarcopenia and osteoporosis. Mice and patients with Cushing syndrome exhibit supraphysiological DBI/ACBP plasma levels. Of note, neutralization of extracellular DBI/ACBP protein with antibodies or mutation of the DBI/ACBP receptor (i.e. the GABRG2 subunit of GABR [gamma-aminobutyric acid type A receptor]) renders mice resistant to the induction of Cushing syndrome. Similarly, knockout of Dbi/Acbp in hepatocytes suppresses the corticotherapy-induced surge in plasma DBI/ACBP concentrations and prevents the manifestation of most of the characteristics of Cushing syndrome. We conclude that autophagy-mediated secretion of DBI/ACBP by hepatocytes constitutes a critical step of the pathomechanism of Cushing syndrome. It is tempting to speculate that stress-induced chronic elevations of endogenous glucocorticoids also compromise human health due to the protracted augmentation of circulating DBI/ACBP concentrations.Abbreviations: DBI/ACBP: diazepam binding inhibitor, acyl-CoA binding protein; GABA: gamma-aminobutyric acid; GABAR: gamma-aminobutyric acid type A receptor; GABRG2: gamma-aminobutyric acid type A receptor subunit gamma2.

DBI/ACBP是一种古老的激素,刺激食欲和脂肪合成代谢。在对饥饿的反应中,DBI/ACBP通过非规范的、巨噬/自噬依赖的途径分泌。生理饥饿反射涉及多种细胞类型在饥饿诱导下分泌DBI/ACBP。DBI/ACBP浓度随后增加,刺激细胞外液的食物摄入。最近,我们观察到糖皮质激素作为内源性应激激素和抗炎药物,在转录水平上调DBI/ACBP表达,刺激肝细胞自噬,从而导致循环DBI/ACBP水平激增。糖皮质激素浓度的长期升高导致一种极端形式的代谢综合征,被称为“库欣综合征”,其临床特征包括贪食、多饮、血脂异常、高胰岛素血症、胰岛素抵抗、脂肪营养不良、内脏肥胖、脂肪变性、肌肉减少和骨质疏松症。小鼠和库欣综合征患者表现出生理上的DBI/ACBP血浆水平。值得注意的是,用抗体中和细胞外DBI/ACBP蛋白或突变DBI/ACBP受体(即GABR [γ -氨基丁酸A型受体]的GABRG2亚基)可使小鼠对库欣综合征的诱导产生抗性。同样,敲除肝细胞中的Dbi/Acbp可抑制皮质治疗引起的血浆Dbi/Acbp浓度激增,并阻止库欣综合征的大部分特征的表现。我们认为,自噬介导的肝细胞分泌DBI/ACBP是库欣综合征发病机制的关键步骤。我们很容易推测,应激诱导的内源性糖皮质激素的慢性升高也会由于循环DBI/ACBP浓度的持续增加而损害人体健康。
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引用次数: 0
Integrating bioengineering, super-resolution microscopy and mechanobiology in autophagy research: addendum to the guidelines (4th edition). 在自噬研究中整合生物工程、超分辨率显微镜和机械生物学:指南增编(第 4 版)。
Pub Date : 2025-03-01 Epub Date: 2024-07-20 DOI: 10.1080/15548627.2024.2379065
Andrea Ravasio, Daniel J Klionsky, Cristina Bertocchi

Recent key technological developments, such as super-resolution microscopy and microfabrication, enabled investigation of biological processes, including macroautophagy/autophagy, with unprecedented spatiotemporal resolution and control over experimental conditions. Such disruptive innovations deepened our capability to provide mechanistic understandings of the autophagic process and its causes. This addendum aims to expand the guidelines on autophagy in three key directions: optical methods enabling visualization of autophagic machinery beyond the diffraction-limited resolution; bioengineering enabling accurate designs and control over experimental conditions; and theoretical advances in mechanobiology connecting autophagy and mechanical processes of the cell. Abbreviation: 3D: three-dimensional; SIM: structured illumination microscopy; STORM: stochastic optical reconstruction microscopy.

最近的关键技术发展,如超分辨率显微镜和微加工技术,使我们能够以前所未有的时空分辨率和对实验条件的控制来研究包括大自噬/自噬在内的生物过程。这些颠覆性创新加深了我们对自噬过程及其原因的机理认识。本增编旨在从三个关键方向扩展有关自噬的指南:光学方法使自噬机制的可视化超越衍射极限分辨率;生物工程使精确设计和控制实验条件成为可能;机械生物学的理论进展将自噬与细胞的机械过程联系起来。缩写:缩写:3D:三维;SIM:结构照明显微镜;STORM:随机光学重建显微镜。
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引用次数: 0
ATP6V1D drives hepatocellular carcinoma stemness and progression via both lysosome acidification-dependent and -independent mechanisms. ATP6V1D 通过溶酶体酸化依赖性和非依赖性机制驱动肝细胞癌的干性和进展。
Pub Date : 2025-03-01 Epub Date: 2024-10-10 DOI: 10.1080/15548627.2024.2406186
Zhijie Xu, Ruiyang Liu, Haoying Ke, Fuyuan Xu, Pengfei Yang, Weiyu Zhang, Yi Zhan, Zhiju Zhao, Fei Xiao

Metabolic reprogramming is pivotal in cancer stem cell (CSC) self-renewal. However, the intricate regulatory mechanisms governing the crosstalk between metabolic reprogramming and liver CSCs remain elusive. Here, using a metabolic CRISPR-Cas9 knockout screen, we identify ATP6V1D, a subunit of the vacuolar-type H+-translocating ATPase (V-ATPase), as a key metabolic regulator of hepatocellular carcinoma (HCC) stemness. Elevated ATP6V1D expression correlates with poor clinical outcomes in HCC patients. ATP6V1D knockdown inhibits HCC stemness and malignant progression both in vitro and in vivo. Mechanistically, ATP6V1D enhances HCC stemness and progression by maintaining macroautophagic/autophagic flux. Specifically, ATP6V1D not only promotes lysosomal acidification, but also enhances the interaction between CHMP4B and IST1 to foster ESCRT-III complex assembly, thereby facilitating autophagosome-lysosome fusion to maintain autophagic flux. Moreover, silencing CHMP4B or IST1 attenuates HCC stemness and progression. Notably, low-dose bafilomycin A1 targeting the V-ATPase complex shows promise as a potential therapeutic strategy for HCC. In conclusion, our study highlights the critical role of ATP6V1D in driving HCC stemness and progression via the autophagy-lysosomal pathway, providing novel therapeutic targets and approaches for HCC treatment.Abbreviations: 3-MA: 3-methyladenine; ANT: adjacent normal liver tissues; ATP6V1D: ATPase H+ transporting V1 subunit D; BafA1: bafilomycin A1; CHMP: charged multivesicular body protein; co-IP: co-immunoprecipitation; CSC: cancer stem cell; ESCRT: endosomal sorting complex required for transport; HCC: hepatocellular carcinoma; IF: immunofluorescence; IHC: immunohistochemical; LCSCs: liver cancer stem cells; qRT-PCR: quantitative real time PCR; V-ATPase: vacuolar-type H+- translocating ATPase; WB: western blot.

代谢重编程是癌症干细胞自我更新的关键。然而,新陈代谢重编程与肝脏干细胞之间错综复杂的调控机制仍然难以捉摸。在这里,我们利用代谢CRISPR-Cas9基因敲除筛选,发现空泡型H+转运ATP酶(V-ATP酶)的一个亚基ATP6V1D是肝细胞癌(HCC)干性的一个关键代谢调节因子。ATP6V1D 表达升高与 HCC 患者的不良临床预后相关。在体外和体内敲除 ATP6V1D 可抑制 HCC 干性和恶性进展。从机理上讲,ATP6V1D通过维持大自噬/自噬通量增强了HCC的干性和进展。具体来说,ATP6V1D不仅能促进溶酶体酸化,还能增强CHMP4B和IST1之间的相互作用,促进ESCRT-III复合物的组装,从而促进自噬体-溶酶体融合,维持自噬通量。此外,沉默CHMP4B或IST1可减轻HCC的干性和进展。值得注意的是,以V-ATP酶复合物为靶点的低剂量巴佛洛霉素A1有望成为HCC的一种潜在治疗策略。总之,我们的研究强调了ATP6V1D在通过自噬-溶酶体途径驱动HCC干性和进展中的关键作用,为HCC治疗提供了新的治疗靶点和方法。
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引用次数: 0
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Autophagy
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