FUNCTIONAL ACTIVITY OF ANTI-GD2 CAR-T CELLS WITH DIFFERENT ANTIGEN-RECOGNITION MODULE

Abstract

Neuroblastoma (NB) is the most common extracranial solid tumor in children, 8-10% of all pediatric tumors and an incidence of about 1-1.3 cases per 100,000 children under 15 years of age. Despite the use of intensive treatment with surgery, high-dose chemotherapy and radiotherapy, the 5-year event-free survival rate is 25-50% and 10-40% after relapse. Over the past decade, a new type of cell therapy with chimeric antigen receptor (CAR) modification of lymphocytes has been rapidly developed. One of the main known antigens for the development of CAR-T therapy against neuroblastoma is the disialoganglioside GD2, the expression of which is characterized in 100% of cases of this disease. Most clinical anti-GD2 CAR variants are based on scFv 14.G2, originating from chimeric antibody 14.18 (denutuximab). In 2020, the FDA approved a new anti-GD2 humanized antibody, 3F8 (naxitamab) with a better safety profile.  Despite partial success, the results of anti-GD2 CAR-T therapy remain modest. One option to increase receptor specificity is targeting O-acetyl-GD2, a disialoganglioside derivative in which the external sialic acid residue is modified with an O-acetyl ester. Acetylation of GD2 occurs only in tumor cells and is not found in peripheral nerves. An 8B6 antibody is known to target O-acetyl-GD2. Thus, at least 3 therapeutic antibodies, 14G2a, hu3F8 and 8B6 compete with each other for targeting GD2 with CAR-T cells. And in all cases the anti-GD2 CAR contains insertion domains in the extracellular part of the molecule. Results of separate clinical trials have been published for CAR-T based on 14G2a and hu3F8, but there are no data on the use of the 8B6 antibody in CARs yet. The aim of the present study is to obtain chimeric antigenic receptors of the 2nd generation based on three antibodies, with different lengths of the extracellular domain and to evaluate their functional activity against a number of cell lines to justify further clinical trials.

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