FREE RADICAL BIOLOGY AND MEDICINE
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2025
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IF 8.2
Zhiyu Qian,Xinyue Zhang,Jiahua Huang,Yumin Hou,Chunlan Hu,Yirui Cao,Nannan Wu,Tongyu Zhu,Guoyi Wu
Renal ischemia-reperfusion injury (IRI), a common complication following kidney transplantation and partial nephrectomy, is the leading cause of renal dysfunction with limited treatment. Excessive cellular iron accumulation drives lipid peroxidation and activates pathways associated with ferroptosis, which has been implicated in renal IRI. However, the regulatory mechanisms of cellular iron metabolism and its relationship with ferroptosis during ischemia-reperfusion (IR) remain unclear. In this study, in vitro OGSD-R (oxygen, glucose, and serum deprivation-restoration) models and in vivo IR models were employed to investigate alterations in iron metabolism, ferroptosis, and the underlying molecular mechanisms using immunofluorescence, immunoblotting and biochemical testing. We identified glucose deprivation-restoration (GD-R) as a key trigger of cellular iron overload under renal IR condition. Mechanistically, GD-R-induced iron overload is driven by the dysfunction of vacuolar ATPase (V-ATPase)-mammalian target of rapamycin (mTOR) pathway. Inactivation of mTOR results in lysosomal iron releases via two-pore channel 2 (TPC2) and ferritin degradation through ferritinophagy. This process elevates intracellular iron levels, thereby promoting ferroptosis in renal IRI. Targeting cellular iron metabolism effectively alleviates renal IRI. These findings highlight the critical role of glucose metabolism and V-ATPase-mTOR pathway in the regulation of iron homeostasis and ferroptosis during renal IRI, and establish a mechanistic link among glucose metabolism, iron overload and ferroptosis, providing potential therapeutic targets for renal IRI.
研究领域:
医学
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细胞生物学
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 8
·4493
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IF 4.9
Jiuge Tang,Dongmei Fang,Jialing Zhong,Min Li
Canonical autophagy is an evolutionarily conserved process that forms double-membrane structures and mediates the degradation of long-lived proteins (LLPs). Noncanonical autophagy (NCA) is an important alternative pathway involving the formation of microtubule-associated protein 1 light chain 3 (LC3)-positive structures that are independent of partial core autophagy proteins. NCA has been defined by the conjugation of ATG8s to single membranes (CASM). During canonical autophagy and NCA/CASM, LC3 undergoes a lipidation modification, and ATG16L1 is a crucial protein in this process. Previous studies have reported that the WDR domain of ATG16L1 is not necessary for canonical autophagy. However, our study found that WDR domain deficiency significantly impaired LLP degradation in basal conditions and slowed down LC3-II accumulation in canonical autophagy. We further demonstrated that the observed effect was due to a reduced interaction between ATG16L1 and FIP200/WIPI2, without affecting lysosome function or fusion. Furthermore, we also found that the WDR domain of ATG16L1 is crucial for chemical-induced NCA/CASM. The results showed that removing the WDR domain or introducing the K490A mutation in ATG16L1 significantly inhibited the NCA/CASM, which interrupted the V-ATPase-ATG16L1 axis. In conclusion, this study highlights the significance of the WDR domain of ATG16L1 for both canonical autophagy and NCA functions, improving our understanding of its role in autophagy.
研究领域:
细胞生物学
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分子生物学
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自噬
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PubMed
高影响因子
Advanced Science
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2025
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IF 14.1
Yu-Juan Zhan,Chun-Miao Deng,Lin Tang,Shu-Jun Li,Tao-Yang Xu,Xian Wei,Xin-Yi Zhang,Can-Can Zheng,Li Deng,Cui Shao,Zhong-Min Ouyang,Alfred King-Yin Lam,Rong Zhang,Jun Liu,Xing-Yuan Shi,Zhen-Yu Pan,Wei Dai,Ming-Liang He,Simon Law,Xu Li,Xiao-Bing Chen,Cheng Zhou,Bin Li,Wen-Wen Xu
N-acetyltransferase 10 (NAT10)-catalyzed N4-acetylcytidine (ac4C) modification has been reported to drive tumor metastasis. Lysosomal dysregulation plays an important role in human diseases, but its function in esophageal cancer metastasis is unclear. It remains unknown whether NAT10 regulates lysosomal function, and the underlying mechanism and treatment strategy warrants investigation. Here, a novel role of NAT10 in inducing lysosomal acidification is revealed, and the clinical and biological significance of ATP6V0E1 in tumor metastasis is uncovered. Mechanistically, NAT10 promotes the translation efficiency of ATPase H + transporting V0 subunit e1 (ATP6V0E1) mRNA in an ac4C-dependent manner to facilitate ATP6V0E1 expression and vacuolar H + -ATPase (v-ATPase) activity, enhancing the lysosomal degradation of E-cadherin, ultimately accelerating tumor metastasis. Furthermore, G-749 is screened and identified as a novel NAT10 inhibitor capable of effectively impeding lysosomal acidification and tumor metastasis by disrupting the NAT10-Ubiquitin-specific Peptidase 39 (USP39) interaction. Overall, the results unveil a novel role of ac4C modifications in regulating lysosomal acidification and propose a potential strategy by targeting NAT10 to inhibit esophageal cancer metastasis.
研究领域:
癌症生物学
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分子生物学
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生物化学
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细胞生物学
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肿瘤生物学
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RNA修饰
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药理学
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高影响因子
Autophagy
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2025
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IF 14.3
Jiahui Long,Huazhong Xie,Hao Shan,Ruoqing Chen,Jialing Zhong,Jiuge Tang,Dongmei Fang,Yao Wang,Peiqing Liu,Xiao-Ming Yin,Min Li
Formation of MAP1LC3/LC3 (microtubule associated protein 1 light chain 3)-positive structures that does not require all of the core ATG (autophagy related) proteins is emerging in the process of noncanonical autophagy (NCA). While LC3 lipidation on endolysosomal membranes has been well characterized, the involvement of other membrane sources and the regulatory mechanisms governing LC3 lipidation in alternative forms of NCA remain poorly understood. Here, we demonstrate the occurrence of LC3 lipidation on the trans-Golgi network (TGN) platform. Different from canonical autophagosomes, these LC3-positive structures do not fuse with lysosomes, and fail to degrade long-lived proteins. In addition, the functional vacuolar-type H+-translocating ATPase (V-ATPase)-ATG16L1 axis is found to be essential for TGN-associated NCA. Notably, in this process, the cytosolic but not lysosomal V1 complex of the V-ATPase assembles at the TGN and plays a pivotal role in further induction of NCA. Eventually, IL1B/IL-1β (interleukin 1 beta) secretion is found to be efficiently enhanced by such TGN-associated NCA, independently of GSDM (gasdermin)-mediated pore formation. Thus, besides the known endolysosome-related NCA, we identify a distinct form of TGN-associated NCA mediated by the V-ATPase-ATG16L1 axis. Such NCA might work as a protein-transport route for the extracellular secretion of IL1B, revealing a mechanism linking Golgi-derived NCA to inflammatory cytokines release.
研究领域:
细胞生物学
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免疫学
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自噬研究
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炎症
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膜运输
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PubMed
高影响因子
Advanced Science
·
2026
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IF 14.1
Lang Chen,Wenyi Lin,Lang Jiang,Xuehui Gao,Chenyan Yu,Jili Han,Zhenping Liu,Xuefeng Li,Mei He,Chang Li,Qilan Li,Chaoying Song,Jiqian Xu,You Shang
Patients with sepsis exhibit circadian disruption and persistent immunosuppression. However, the molecular mechanisms linking them remain unclear. Integration of multi‐cohort transcriptomic and single‐cell datasets shows that circadian gene dysregulation in patients with sepsis and septic mice correlates with disease severity and immunosuppressive states, with monocytes/macrophages emerging as a principal affected population. Sustained endotoxin stimulation elevates the core clock repressor NR1D1 in macrophages, which occupies the Igf2bp2 promoter and suppresses its transcription. Loss of IGF2BP2 destabilizes the V‐ATPase subunit transcripts Atp6v1b2 and Atp6v0c through an m 6 A‐dependent mechanism, disrupting phagolysosomal acidification rhythms and pathogen clearance. siRNA‐mediated NR1D1 knockdown restores IGF2BP2 expression, circadian oscillations, and phagolysosomal function during the development of endotoxin tolerance. To achieve therapeutic delivery, we engineer hybrid membrane nanovesicles (siNR1D1@HM‐LNP) that reverse circadian and immune dysregulation in septic mice, enhance bacterial clearance, and markedly improve survival. These findings establish an NR1D1‐mediated circadian‐immune coupling mechanism and provide a therapeutic strategy for targeting sepsis‐induced immunosuppression. Sepsis disrupts immune‐cell rhythms and weakens bacterial clearance. Biomimetic nanovesicles combining erythrocyte and inflammation‐activated macrophage membranes deliver siNR1D1 to dysfunctional macrophages, restoring the NR1D1–IGF2BP2–V‐ATPase pathway, circadian regulation, phagolysosomal acidification, and antimicrobial defense. In mouse models of polymicrobial sepsis and secondary pneumonia, treatment improves immune balance and bacterial control, reduces organ injury, and increases survival.
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高影响因子
Autophagy
·
2026
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IF 18.6
Zhengran Li,Huiqing Zhao,Danjie Zhang,Jing Wei,Xiyue Tao Liu,Ruiyan Kong,Xuejing Ren,Hang Zhao,Xiaomin Fang,Jingyan Fu,Zhouhua Li
The self-renewal and differentiation of stem cells are tightly controlled to maintain tissue homeostasis. Failure in stem cell maintenance results in stem cell depletion and precocious aging. However, how stem cells are maintained still remains not fully understood. Here, through a large-scale RNAi screen for maintenance and proliferation of adult Drosophila intestinal stem cells (ISCs), we identify several subunits of V-ATPase including Vha68-2/ATP6V1A, required for ISC proliferation, differentiation and tissue regeneration. Inactivation of Vha68-2 results in accumulation of plasma membrane (PM)-derived structures in autophagosomes and lysosomes through Atg16 and Rab5. Furthermore, Vha68-2-defective ISCs undergo direct differentiation due to ectopic Notch activation. Mechanistically, Vha68-2 facilitates Notch receptor internalization and subsequent degradation in autolysosomes to quench ectopic Notch activation, thereby maintaining ISC fate. Truncated Notch product devoid of its extracellular domain is accumulated in autolysosomes upon Vha68-2 deficiency. The Notch products accumulated in autolysosomes still require further cleavage to drive ISC differentiation. The functions of Vha68-2 in macroautophagy/autophagy and ISC maintenance are evolutionarily conserved. Our results provide new insights into the underlying mechanism of how autophagy is involved in stem cell maintenance under physiological conditions.Abbreviations: aph-1: anterior pharynx defective 1; ATP6V1A: ATPase H+ transporting V1 subunit A; CASM: conjugation of ATG8s to single membranes; EB: enteroblast; EC: absorptive enterocyte; ER: endoplasmic reticulum; ISC: intestinal stem cell; kuz: kuzbanian; LTDR: LysoTracker Deep Red; LTR: LysoTracker Red; MARCM: mosaic analysis with a repressible cell marker; NECD: Notch extracellular domain; NICD: Notch intracellular domain; PM: plasma membrane; PAS: phagophore assembly site; SNARE: soluble N-ethylmaleimide-sensitive factor-attachment protein receptor; TEM: transmission electron microscopy; TM: tunicamycin.
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高影响因子
Autophagy
·
2026
·
IF 14.3
Hongtao Tie,Mengqian Hou,Jun Zhang,Martijn F. Hoes,Dietbert Neumann,Joost J. F. P. Luiken,Shujin Wang
The vacuolar-type H+-translocating ATPase (V-ATPase) plays a pivotal role in cellular homeostasis by acidifying endosomes and lysosomes, regulating key processes such as autophagy and membrane trafficking. While the importance of V-ATPase in these functions is well-established, the methodologies for studying its assembly and function remain varied and under-characterized. In this study, we systematically validated and compared methodologies for assessing V-ATPase assembly and endo/lysosomal acidification under physiological and high-fat conditions, both in vitro and in vivo. Various techniques, including fractionation, immunoprecipitation, immunofluorescence microscopy, and proximity ligation assays, were evaluated using cardiomyocyte cell lines, rat models of lipid overload, and two heart-specific V-ATPase-knockout mouse models (V-ATPase subunits ATP6V1G1 and ATP6V0D2). High palmitate (HP) and bafilomycin A1 (BafA) were used to manipulate v-ATPase function, while a colorimetric assay assessed proton-pumping activity. Results consistently showed that HP and BafA induced V-ATPase disassembly and inhibited proton-pumping activity, leading to impaired endo/lysosomal acidification and autophagy inhibition upon fusion of autophagosomes with lysosomes. Similar findings were observed in vivo, where a high-fat diet (HFD) reproduced the effects of HP on cardiac tissue. The methodologies were further validated in two heart-specific V-ATPase-knockout mouse models, demonstrating consistent outcomes across different experimental approaches. This study establishes a robust framework for evaluating V-ATPase assembly and function. The validated methodologies reveal that lipid overload inhibits autophagy and contributes to insulin resistance by inducing V-ATPase disassembly and subsequent lysosomal dysfunction. These findings offer insights into the molecular mechanisms underlying metabolic diseases and provide valuable tools for further research.
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高影响因子
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