Összes szerző

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Lima Maria Eduarda

az alábbi absztraktok szerzői között szerepel:

Ghofrane Medyouni
Role of ion channels in CAR T-cell

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P42

Role of ion channels in CAR T-cell

Ghofrane Medyouni¹, Vivien Jusztus¹, Orsolya Vörös¹, Maria Eduarda Lima^1,2, György Panyi¹, Péter B. Hajdu^{1, 2}

¹ University of Debrecen, Faculty of Medicine, Department of Biophysics and Cell Biology

² University of Debrecen, Faculty of Dentistry, Division of Dental Biochemistry

Cancer immunotherapy partly relies on the reprogramming of host immune cells to eliminate cancer cells. Genetic modification of T cells to express chimeric antigen receptors (CARs) is utilized in the treatment of hematological malignancies. Despite its success, many challenges remain to improve the efficacy and safety of this therapy. Ion channels in T-cells participate in the regulation of Ca²⁺-dependent activation pathway and play a role in various effector functions inevitable for target cell abolition. Hence, modification of ion channels’ function can contribute to successful immune therapy. However, no study has been reported about functional role of CAR T-cell ion channels yet.

We established a 3^rd-generation CAR expressing cell line (CD19-CAR cells) from Jurkat cells. We used the whole-cell patch-clamp technique and FURA-2-based Ca²⁺-imaging to determine the biophysical properties of Kv1.3 and Ca²⁺-response of CD19-CAR cells, respectively. We adapted a Calcein Red based killing assay to test CD19-CAR cells’ target cell killing capacity. We assessed the localization of Kv1.3 in standalone and in the CAR-synapse engaged CD19-CAR cells.

We showed that Kv1.3 activation and inactivation kinetics are the same in non-transfected and CD19-CAR cells, while voltage-dependence of activation was different. Thapsigargin-induced Ca²⁺-response of CD19-CAR cells was lower as compared to the control. We showed that Kv1.3 channels are co-localized with CARs in standalone CD19-CAR cells, and they redistribute to the contact region between a CD19-CAR cell and a target cell (Raji B cell). Upon Vm24 addition (specific Kv1.3 inhibitor, 1 nM) the target cell ability of CD19-CAR cells was impaired. Based on these results, we suppose that ion channels can affect the outcome of immunotherapy, and further experiments are needed to clarify their functional role.

Acknowledgment

This work was supported by Stipendium Hungaricum Scholarship to M.G.t and NKFIH (K128525, P.H.).