Magnetic bead-sensitized optoporation coupled with antibodies-based activation for mRNA CAR-T cell manufacturing

Article Article scientifique Article Magnetic bead-sensitized optoporation coupled with antibodies-based activation for mRNA CAR-T cell manufacturing eng 2025-02-04 2025-03-13 author Maldonado-Perez Noëlia Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland. 412875 author Doucey Marie-Agnès Limula SA, 1066 Épalinges, Switzerland. author Dzhumashev Dzhangar Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland. 411024 Département de médecine (DEMED) author Martinez Bedoya Darel Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland. 932040 Département de médecine (DEMED) author Castillo Cantero Luis Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland. 983983 Département de médecine (DEMED) author Boudousquie Caroline Limula SA, 1066 Épalinges, Switzerland. author Pierson Yann Limula SA, 1066 Épalinges, Switzerland. author Henry Luc Limula SA, 1066 Épalinges, Switzerland. author Dutoit Vallotton Valérie 0000-0002-3469-4237 Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland. 668630 Département de médecine (DEMED) author Migliorini Denis Center for Translational Research in Onco-Hematology, University of Geneva, 1211 Geneva, Switzerland.,Department of Oncology, University Hospital of Geneva, 1205 Geneva, Switzerland.,Swiss Cancer Center Léman, Lausanne, Switzerland.,Agora Cancer Research Center, 1005 Lausanne, Switzerland. 838158 Département de médecine (DEMED) 101428 10.1016/j.omtm.2025.101428 40092640 2329-0501 PMC11910140 https://www.sciencedirect.com/science/article/pii/S2329050125000233 Immunotherapy is facing a revolution with the advent of immune cell engineering. Chimeric antigen receptor (CAR)-T cell therapy has shown unprecedented efficacy in B cell malignancies and is now being evaluated in other disease areas. Viral transduction is the most common method for immune cell genetic engineering, but presents important limitations, such as high reagent costs and regulatory concerns due to mutagenesis risk. One prevailing non-viral gene delivery strategy relies on the electroporation of non-integrating RNA. However, most modern electroporation technologies also require high reagent costs and rely on the use of proprietary software and transfection buffers. Nanoparticle-sensitized optoporation represents an alternative method for transient permeabilization of cells. Here, we introduce magnetic bead-sensitized optoporation, in which commercially available superparamagnetic beads coupled with anti-human CD3 and CD28 antibodies are used as photosensitizers for efficient genetic cargo delivery into human primary T cells and other immune cells. We show that magnetic bead-sensitized optoporation of human T cells generates functional mRNA-based CAR-T cells without affecting T cell product memory phenotype or activation potential. Importantly, optoporated T cells exhibited a greater proliferation capacity relative to electroporated T cells. In conclusion, our findings suggest that magnetic bead-sensitized optoporation holds promise as mRNA delivery strategy for immune cell therapy. CAR-T cells Cell therapy Electroporation MRNA Magnetic beads Optoporation Dewey 610 Dewey 616 56 Département de médecine 221 4f7589ed-e44d-4850-a4d8-b0c1eccdfe48 Centre de recherche translationnelle en onco-hématologie 209 false 1310 Tumeurs cérébrales 1016 Molecular therapy. Methods & clinical development 33 1 a4a25144-b9a6-41a6-a02e-5c68958ec23d Article (Published version) Public Maldonadomolther2025.pdf 4215652 application/pdf CC BY-4.0 1605742b2eda0f9b70a7f07dbe2702c5e307cf7fad4f47163cd264b656ec8031 7da35f9c-6199-46f2-ac73-2b235a36e4b3 Supplemental data Public 1-s2.0-S2329050125000233-mmc1.pdf 1443748 application/pdf 4985d0e443973a4d20cc3423cb3b5dd5ec4d3d330469c7bde9a6356f3560640b openAccess Faculté de médecine / Section de médecine clinique / Département de médecine Faculté de médecine / Centre de recherche translationnelle en onco-hématologie

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Magnetic bead-sensitized optoporation coupled with antibodies-based activation for mRNA CAR-T cell manufacturing	Molecular therapy. Methods & clinical development			2025	63	175

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Dzhumashev, Dzhangar