Salinas, María DoloresMartínez, Carlos M.Roca, Francisco J.García-Bernal, DavidMartínez-Morga, MartaRodríguez-Madoz, Juan R.Prósper, FelipeZapata González, Agustín GregorioMoraleda, Jose MaríaMartínez, SalvadorValdor, Rut2025-10-222025-10-222025-04-24Salinas, M. D., Martínez, C. M., Roca, F. J., García-Bernal, D., Martínez-Morga, M., Rodríguez-Madoz, J. R., Prósper, F., Zapatak, A. G., Moraleda, J. M., Martínez, S., & Valdor, R. (2025). Chaperone-mediated autophagy sustains pericyte stemness necessary for brain tissue homeostasis. Journal of Advanced Research. Advance online publication. https://doi.org/10.1016/j.jare.2025.04.0152090-123210.1016/j.jare.2025.04.015https://hdl.handle.net/20.500.14352/125263This work was mainly developed by Seneca 20840/PI/18 funded by Seneca Foundation 'Agencia de Ciencia y Tecnología de la Región de Murcia', by Ramon y Cajal (RYC) 2019-027520-I funded by Ministerio de Ciencia, Innovación y Universidades (MCIU) and Agencia Estatal de Investigación (AEI) MCIU/AEI/10.13039/50110-0011033, as 'European Social Fund (ESF) Investing in your future', and by PID2020-114010RB-100 and PID2023-1491110B-100 funded by MICIU/AEI/10.13039/50110001 1033 and Fondo Europeo de Desarrollo Regional (FEDER), UE (to RV). It was also partially supported by Caixalmpulse CI23-20487 funded by 'La Caixa' Foundation (to RV) and by Spanish Network of Advanced Therapies (TERAV), RICORS subprogram (to AGZ, FP, SM and JMM), funded by Instituto de Salud Carlos III and co-funded by European Regional Development Fund (ERDF)-Next Generation EU 'Plan de Recuperación, Transformación y Resiliencia'. FJR was supported by fellowship RYC2019-571 027799-1 funded by MCIU/AEI. We thank Jose Luis Ferran at Human Anatomy Department, UMU and Pablo Pelegrín's lab at Experimental Surgery Unit, IMIB for his always kind help providing tools for this study. We thank Professor Ana Maria Cuervo at the Developmental Molecular Biology Department and Professor Fernando Macian at the Pathology Department of Albert Einstein College of Medicine (New York, United States) for all their kind help providing the deficient CMA mice and the CMA reporter and in the initial design of this study and data discussion. We thank the Pathology, Bioinformatic and Animal facilities of IMIBIntroduction Pericytes (PCs) are mural cells exhibiting some mesenchymal stem cell (MSC) properties and contribute to tissue regeneration after injury. We have previously shown that glioblastoma cancer cells induce in PCs, a pathogenic upregulation of chaperone-mediated autophagy (CMA) which modulates immune functions and MSC-like properties to support tumor growth. Objectives The aim of the study was to interrogate the role of CMA-regulated MSC properties in PCs in the context of tissue repair during inflammation triggered by a demyelinating injury. Methods Studies of RNA-seq were done PCs with (WT) and without (LAMP-2A KO) CMA. Cell characterization related to stemness, lineage and morphology was done in WT and KO PCs. Secretome analysis and cell differentiation assay using the supernatants from CMA-efficient and deficient PCs cultures was done in mesenchymal cells. Inflammatory response of brain cells was assessed with WT and KO PCs secretome. To corroborate in vitro results, CMA modulation in response to inflammation in PCs and tissue repair markers were measured in the lesion areas of a demyelination mouse model and correlated with the tissue reparation after intravenous PC administration. An inflammatory mediator was used to study effects on PC-CMA activity. Results We found that inflammatory mediators such as IFNγ downregulate CMA in PCs, suppressing PC stemness and promoting a pro-inflammatory secretome. Restoration of PC CMA activity during inflammation maintains PC MSC properties and induces an MSC-like proteome which decreases inflammation and promotes tissue repair. We identified secreted proteins involved in regenerative and protective processes, and therefore, necessary to restore brain tissue homeostasis after inflammation induced by a demyelinating injury. Conclusion we show that manipulation of CMA activity in host PCs could be a useful therapeutical approach in the context of brain inflammation, which might be extended to other diseases where the pericyte has a key role in response to inflammation.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal articlehttps://doi.org/10.1016/j.jare.2025.04.015open access577.1577.2611.018612.8Chaperone-mediated autophagyIFNγInflammatory damageInjuryPericytes as functional mesenchymal stem cellsSecretomeBiología celular (Biología)Biología molecular (Biología)Neurociencias (Biológicas)InmunologíaBioquímica (Biología)2403 Bioquímica2407 Biología Celular2302.21 Biología Molecular2490 Neurociencias