Towards minimally invasive delivery of large molecules into human T cells by photoporation with polydopamine nanosensitizers and photothermal nanofibers
Adoptive T cell transfer has demonstrated remarkable clinical success in the treatment of hematological malignancies, holding great potential to transform cancer care. Development of such novel therapies requires ex vivo T cell engineering with a synthetic tumor antigen-specific receptor. To address the complexities and high costs associated with viral vector-based T cell manufacturing, alternative non-viral approaches have been actively investigated to transfect lymphocytes with exogenous genetic material. Photoporation is a physical intracellular delivery method that combines photo-responsive nanomaterials and laser irradiation to achieve transient membrane permeabilization by distinct photothermal effects. In this dissertation, we investigated the applicability of photoporation for intracellular delivery of model FITC-dextran macromolecules in unstimulated and stimulated human T cells, as well as the functional implications of photoporation treatment. We first synthesized polydopamine nanoparticles as a biocompatible and biodegradable alternative to traditionally used metallic photosensitizers, showing effective macromolecule delivery in both T cell models. Next, we demonstrated the necessity to tailor the nanosensitizer size to cell type to preserve T cell phenotype and functionality post-treatment. Finally, to alleviate safety and regulatory concerns over cell exposure to free nanoparticles, we evaluated a photothermal system, where iron oxide nanoparticles were embedded within polycaprolactone nanofiber meshes fabricated by electrospinning, showing effective model macromolecule delivery in both T cell phenotypes. Altogether, the data in this work provide important considerations for further optimization of photoporation technology for T cell engineering towards clinically relevant applications.
Ghent : Ghent University & University of Antwerp , 2023
234 p.
Supervisor: Braeckmans, Kevin [Supervisor]
Supervisor: De Vos, Winnok [Supervisor]
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Creation 18.10.2023
Last edited 21.10.2023
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