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Neuronal LAMP2A-mediated reduction of adenylyl cyclases induces acute neurodegenerative responses and neuroinflammation after ischemic stroke

Cell Death & Differentiation volume 32, pages 337–352 (2025)Cite this article

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Abstract

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.

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Fig. 1: Neuronal Lamp2 gene deletion alleviates cerebral infarction and long-term neural deficits in mice after ischemic stroke.
Fig. 2: CMA pathway is overactivated and LAMP2A deficiency lessens molecular levels of PANoptosis-like cell death in primary cortical neurons after OGD/R.
Fig. 3: LAMP2A deficiency prevents against post-stroke PANoptosis-like molecular pathway of neuronal death and glial activation in mice.
Fig. 4: Proteome profiles of LAMP2A deficiency in cortical neurons after acute metabolic stress.
Fig. 5: PKA inhibitor H89 rekindles post-stroke PANoptosis-like molecular pathway of neuronal death under conditions of LAMP2A deficiency in mice.
Fig. 6: Neuronal LAMP2A-lysosome pathway is responsible for the degradation of ADCY1 and ADCY3 in mice after ischemic stroke.
Fig. 7: Inducers of the PKA signaling pathway suppress neuronal PANoptosis-like responses and glial activation in mice following stroke.
Fig. 8: Inducers of the PKA signaling pathway alleviate cerebral infarction and long-term neurocognitive deficits in mice following stroke.

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Data availability

The original data in the study are included in this article or the supplementary material. Additional data are available upon reasonable request.

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Acknowledgements

The authors gratefully acknowledge Ms. Jie Zhao (Animal Experimental Center of Public Experimental Platform, China Pharmaceutical University) for her assistance with behavioral experiments.

Funding

This study was supported by grants from the National Natural Science Foundation of China (No. 82173801 and 81673418).

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Authors and Affiliations

  1. State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, China

    Dingfang Shi, Yunhao Bai, Ruiling Long, Jing Xia, Wenxuan Xu, Dongshen Qin, Xuejun Yang, Ming Ding & Xiao-Yu Hou

  2. School of Life Science and Technology, China Pharmaceutical University, Nanjing, Jiangsu, China

    Dingfang Shi, Yunhao Bai, Ruiling Long, Jing Xia, Wenxuan Xu, Dongshen Qin, Xuejun Yang, Ming Ding & Xiao-Yu Hou

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  1. Dingfang Shi
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Contributions

D.S., Y.B. performed biochemical and molecular biology experiments, lysosomal isolation and substrate analysis. R.L. and M.D. performed quantitative proteomic analysis. D.S., J.X. and W.X. performed experiments on primary neurons. D.S., Y.B., D.Q. and X.Y. performed neurofunctional assessment and behavioral experiments. D.S., Y.B. and R.L. conducted data analysis and visualization. M.D. and X.Y.H. planned the experimental design. X.Y.H. conceived and supervised the project. D.S., W.X. and X.Y.H. wrote the paper with feedback from all authors.

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Correspondence to Ming Ding or Xiao-Yu Hou.

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All experiments were approved by the Institutional Animal Care and Use Committee of China Pharmaceutical University and were conducted in accordance with the ARRIVE guidelines.

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Shi, D., Bai, Y., Long, R. et al. Neuronal LAMP2A-mediated reduction of adenylyl cyclases induces acute neurodegenerative responses and neuroinflammation after ischemic stroke. Cell Death Differ 32, 337–352 (2025). https://doi.org/10.1038/s41418-024-01389-0

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