Toxicology Research
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2025
·Vol. 14
·期 2
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IF 2.1
Fu Dan,Li Ziyou,Feng Huafeng,Fan Fangling,Zhang Wang,He Liang
Spinal cord injury (SCI)-induced ischemic delayed paralysis is one of the most serious side effects of aneurysms surgeries. Recent studies prove that the activation of autophagy, including macroautophagy and micro-autophagy pathways, occur during SCI-induced brain neuron damage. However, the role of chaperone mediated autophagy (CMA) during SCI remains to be unveiled. In the present work, rat model of delayed paralysis after aneurysms operation and adenovrius induced LAMP2A knockdown in microglia cells were applied in the present work to investigate the involvement of LAMP2A-mediated CMA in the aneurysm operation related SCI and delayed paralysis. The results showed that LAMP2A was upregulated in the SCI procedure, and contributed to neuron death and pro-inflammation perturbation via inducing iNOS+ polarization in microgila. We additionally observed that knockdown of LAMP2A resulted in the shift of microglia from iNOS+ to ARG1+ phenotype, as well as alleviated neuron damage during SCI. Furthermore, the analysis of BBB score, the result of immunohistological staining, and protein detection confirmed the activation of LAMP2A-mediated CMA activation and its interaction with NF-κB signaling, which leads to neuron death and motor function loss. These results prove that LAMP2A-mediated CMA contributes to the upregulation of pro-inflammatory cytokines and results in cell death in neurons during ischemic delayed paralysis via activating NF-κB signaling. Inhibition of LAMP2A promotes neurons survival during ischemic delayed paralysis.
研究领域:
神经科学
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分子生物学
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医学
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2025
·Vol. 26
·期 6
·2804
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IF 4.9
Danni Chen,Opeyemi B. Fasina,Jiahui Lin,Jiayuan Zeng,Majid Manzoor,Hiroshi Ohno,Lan Xiang,Jianhua Qi
In previous studies, we isolated a series of novel gentisides with nerve growth factor (NGF)-mimic activities fromGentiana rigescensFranch and conducted continuous structure–activity relationship (SAR) studies. Recently, a lead compound named TBG096 was discovered with significant NGF-mimic activity, low toxicity, and ability to pass through the blood–brain barrier (BBB). At the cell level, TBG096 exerts NGF-mimic activity by regulation of heat-shock cognate protein 70 (Hsc70) and downstream proteins. Subsequently, high-fat diet (HFD)-induced Alzheimer disease (AD) mouse models were used to evaluate the anti-AD efficacy of the compound. TBG096 significantly improved the memory dysfunction of AD mice at doses of 0.1, 5, and 20 mg/kg, respectively. In order to elucidate the mechanism of action of the compound against AD, the RNA-sequence analysis of transcriptomics, quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence staining, and Western blot analysis were performed using animal samples. TBG096 significantly increased the expression of the Wnt gene family (Wnt10b,Wnt5a, andWnt1) and the number of mature neurons and newborn neurons in the hippocampus and cerebral cortex of AD mice, respectively. At the same time, it reduced the activity of microglia, astrocyte cells, and expression of inducible nitric oxide synthase (INOS) in the brain. Moreover, this compound significantly increased phosphorylated-adenosine 5′-monophosphate-activated protein kinase (AMPK), Hsc70, and lysosomal-associated membrane protein 2a (LAMP2A) and decreased the expression of hexokinase 2 (HK2), pyruvate kinase M2 (PKM2), amyloid precursor protein (APP), microtubule-associated protein tau (Tau), phosphoryl-Tau, and β-amyloid (Aβ) at the protein level. These results suggest that TBG096 produced the NGF-mimic activity and the anti-AD effect via promoting neurogenesis and modification of the Hsc70/HK2/PKM2/LAMP2A signaling pathway, proposing a potential novel approach to counteracting cognitive decline by developing small molecules that promote neurogenesis and the Hsc70 signaling pathway.
研究领域:
药理学
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神经科学
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分子生物学
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PubMed
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BRITISH JOURNAL OF PHARMACOLOGY
·
2023
·
IF 7.3
Rui-Fang Dong,Cheng-Jiao Qin,Yong Yin,Liang-Liang Han,Cheng-Mei Xiao,Kai-Di Wang,Rong-Yuan Wei,Yuan-Zheng Xia,Ling-Yi Kong
Background and Purpose Chaperone-mediated autophagy (CMA) is a selective type of autophagy targeting protein degradation and maintains high activity in many malignancies. Inhibition of the combination of HSC70 and LAMP2A can potently block CMA. At present, knockdown of LAMP2A remains the most specific method for inhibiting CMA and chemical inhibitors against CMA have not yet been discovered. Experimental Approach Levels of CMA in non-small cell lung cancer (NSCLC) tissue samples were confirmed by tyramide signal amplification dual immunofluorescence assay. High-content screening was performed based on CMA activity, to identify potential inhibitors of CMA. Inhibitor targets were determined by drug affinity responsive target stability-mass spectrum and confirmed by protein mass spectrometry. CMA was inhibited and activated to elucidate the molecular mechanism of the CMA inhibitor. Key Results Suppression of interactions between HSC70 and LAMP2A blocked CMA in NSCLC, restraining tumour growth. Polyphyllin D (PPD) was identified as a targeted CMA small-molecule inhibitor through disrupting HSC70–LAMP2A interactions. The binding sites for PPD were E129 and T278 at the nucleotide-binding domain of HSC70 and C-terminal of LAMP2A, respectively. PPD accelerated unfolded protein generation to induce reactive oxygen species (ROS) accumulation by inhibiting HSC70–LAMP2A–eIF2α signalling axis. Also, PPD prevented regulatory compensation of macroautophagy induced by CMA inhibition via blocking the STX17–SNAP29–VAMP8 signalling axis. Conclusions and Implications PPD is a targeted CMA inhibitor that blocked both HSC70–LAMP2A interactions and LAMP2A homo-multimerization. CMA suppression without increasing the regulatory compensation from macroautophagy is a good strategy for NSCLC therapy. Graphical
研究领域:
肿瘤学
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分子生物学
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药理学
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CELL DEATH AND DIFFERENTIATION
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2024
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IF 13.7
Shi Dingfang,Bai Yunhao,Long Ruiling,Xia Jing,Xu Wenxuan,Qin Dongshen,Yang Xuejun,Ding Ming,Hou Xiao-Yu
Lysosomes regulate cellular metabolism to maintain cell survival, but the mechanisms whereby they determine neuronal cell fate after acute metabolic stress are unknown. Neuron-enriched lysosomal membrane protein LAMP2A is involved in selective chaperone-mediated autophagy and exosome loading. This study demonstrates that abnormalities in the neuronal LAMP2A-lysosomal pathway cause neurological deficits following ischemic stroke and that this is an early inducer of the PANoptosis-like molecular pathway and neuroinflammation, simultaneously inducing upregulation of FADD, RIPK3, and MLKL after ischemia. Quantitative proteomic and pharmacological analysis showed that after acute metabolic stress, the neuronal LAMP2A pathway induced acute synaptic degeneration and PANoptosis-like responses involving downregulation of protein kinase A (PKA) signaling. LAMP2A directed post-stroke lysosomal degradation of adenylyl cyclases (ADCY), including ADCY1 and ADCY3 in cortical neurons. Post-stroke treatment with cAMP mimetic or ADCY activator salvaged cortical neurons from PANoptosis-like responses and neuroinflammation, suggesting that the neuronal ADCY–cAMP–PKA axis is an upstream arrester of the pathophysiological process following an ischemic stroke. This study demonstrates that the neuronal LAMP2A-lysosmal pathway drives intricate acute neurodegenerative and neuroinflammatory responses after brain metabolic stress by downregulating the ADCY–PKA signaling cascade, and highlights the therapeutic potential of PKA signal inducers for improving stroke outcomes.
研究领域:
神经科学
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细胞生物学
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分子生物学
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药理学
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神经病学
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PubMed
高影响因子
Nature Microbiology
·
2026
·Vol. 11
·期 10
·2883-2900
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IF 18.7
Zhang Yulin,Dong Zhongwei,Chen Xiaoting,Peng Ke,Huang Qingye,Rao Wutian,Song Shu,Zhang Liming,Gao Shujun,Zhu Caixia,Wang Yuyan,Robertson Erle,Yuan Zhenghong,Wei Fang,Cai Qiliang
Chaperone-mediated autophagy (CMA) is a critical regulator of cancer progression and antiviral responses, often modulated by post-translational modifications. SUMOylation, a reversible post-translational modification, is implicated in viral infection and tumorigenesis, yet its role in CMA regulation remains unclear. Here we demonstrate that oncogenic viruses hijack CMA by inducing nuclear localization and SUMOylation of lysosomal-associated membrane protein 2A (LAMP2A), a central lysosomal receptor. Specifically, tripartite motif containing 32 (TRIM32) mediates SUMO2/3 conjugation to LAMP2A at lysine residues K104, K132 and K289. Using Kaposi’s sarcoma-associated herpesvirus as a model, we found that TRIM32-induced LAMP2A SUMOylation activates CMA, thereby promoting viral persistence, tumour growth and angiogenesis in vivo. Importantly, TRIM32 expression positively correlates with LAMP2A in tumour tissues from patients with Kaposi’s sarcoma-associated herpesvirus-, Epstein–Barr virus- and human papillomavirus-associated cancers, and peptide inhibitors targeting SUMOylated LAMP2A demonstrate potent antitumour and antiviral efficacy in vitro and in mice. Together, our findings position LAMP2A as a promising druggable target for treating virus-associated cancers.
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高影响因子
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