Survival advantage of native and engineered T cells is acquired by mitochondrial transfer from mesenchymal stem cells

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dc.contributor.authorCourt, Angela C.
dc.contributor.authorParra Crisóstomo, Eliseo
dc.contributor.authorCastro Córdova, Pablo
dc.contributor.authorLorca, Rocío
dc.contributor.authorFigueroa, Fernando E.
dc.contributor.authorKhoury, Maroun
dc.date.accessioned2025-01-22T18:13:39Z
dc.date.available2025-01-22T18:13:39Z
dc.date.issued2024-12
dc.description.abstractBackground: Apoptosis, a form of programmed cell death, is critical for the development and homeostasis of the immune system. Chimeric antigen receptor T (CAR-T) cell therapy, approved for hematologic cancers, retains several limitations and challenges associated with ex vivo manipulation, including CAR T-cell susceptibility to apoptosis. Therefore, strategies to improve T-cell survival and persistence are required. Mesenchymal stem/stromal cells (MSCs) exhibit immunoregulatory and tissue-restoring potential. We have previously shown that the transfer of umbilical cord MSC (UC-MSC)-derived mitochondrial (MitoT) prompts the genetic reprogramming of CD3+ T cells towards a Treg cell lineage. The potency of T cells plays an important role in effective immunotherapy, underscoring the need for improving their metabolic fitness. In the present work, we evaluate the effect of MitoT on apoptotis of native T lymphocytes and engineered CAR-T cells. Methods: We used a cell-free approach using artificial MitoT (Mitoception) of UC-MSC derived MT to peripheral blood mononuclear cells (PBMCs) followed by RNA-seq analysis of CD3+ MitoTpos and MitoTneg sorted cells. Target cell apoptosis was induced with Staurosporine (STS), and cell viability was evaluated with Annexin V/7AAD and TUNEL assays. Changes in apoptotic regulators were assessed by flow cytometry, western blot, and qRT-PCR. The effect of MitoT on 19BBz CAR T-cell apoptosis in response to electroporation with a non-viral transposon-based vector was assessed with Annexin V/7AAD. Results: Gene expression related to apoptosis, cell death and/or responses to different stimuli was modified in CD3+ T cells after Mitoception. CD3+MitoTpos cells were resistant to STS-induced apoptosis compared to MitoTneg cells, showing a decreased percentage in apoptotic T cells as well as in TUNEL+ cells. Additionally, MitoT prevented the STS-induced collapse of the mitochondrial membrane potential (MMP) levels, decreased caspase-3 cleavage, increased BCL2 transcript levels and BCL-2-related BARD1 expression in FACS-sorted CD3+ T cells. Furthermore, UC-MSC-derived MitoT reduced both early and late apoptosis in CAR-T cells following electroporation, and exhibited an increasing trend in cytotoxic activity levels. Conclusions: Artificial MitoT prevents STS-induced apoptosis of human CD3+ T cells by interfering with the caspase pathway. Furthermore, we observed that MitoT confers protection to apoptosis induced by electroporation in MitoTpos CAR T-engineered cells, potentially improving their metabolic fitness and resistance to environmental stress. These results widen the physiological perspective of organelle-based therapies in immune conditions while offering potential avenues to enhance CAR-T treatment outcomes where their viability is compromised.
dc.description.sponsorshipThis work was supported by grants from the Agencia Nacional de Investigaci\u00F3n y Desarrollo\u2014ANID FONDECYT regular #1211749, ANID FONDECYT Iniciaci\u00F3n #11240539, from the Conselho Nacional de Desenvolvimento Cientifico e Tecnol\u00F3gico (CNPq) and the Funda\u00E7\u00E3o Carlos Chagas Filho de Amparo \u00E0 Pesquisa do Estado do Rio de Janeiro (FAPERJ) and ANID-Basal Funding for Scientific and Technological Center of Excellence, IMPACT, #FB210024, Santiago, Chile.
dc.identifier.doi10.1186/s12967-024-05627-4
dc.identifier.issn14795876
dc.identifier.urihttp://repositorio.uandes.cl/handle/uandes/979
dc.language.isoen_US
dc.publisherJournal of Translational Medicine
dc.subjectChimeric Antigen Receptor T (CAR-T) Cell
dc.subjectInduced-apoptosis
dc.subjectMesenchymal Stromal/Stem Cells
dc.subjectMitochondria Transfer
dc.titleSurvival advantage of native and engineered T cells is acquired by mitochondrial transfer from mesenchymal stem cells
dc.typeArticle
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