Development and validation of anti-BCMA CAR-T cells with memory profile for Multiple Myeloma treatment

Abstract

Multiple Myeloma is a common hematological neoplasm, related to high morbidity and mortality. Despite the progress related to better understanding of the disease and its pathophysiology, the disease is still virtually incurable. Normally, the disease presents periods of remission and relapse. Periods of remission tend to become increasingly shorter, and the most frequent and aggressive relapses. Anti-CD19 cell therapy has been shown to be quite effective in the treatment of refractory or recurrent B lymphomas and leukemias. Recently, a possible target for therapy with CAR T-cells designated as anti-BCMA has been identified in neoplastic Multiple Myeloma plasma cells. The main objective of this project is to develop and evaluate the capacity of genetically modified T-lymphocytes that express the anti-BCMA CAR to eradicate Multiple Myeloma neoplastic plasma cells. For this purpose, a second-generation lentiviral vector containing the anti-BCMA clone will be used. Three new BCMA clones, selected using a biopanel methodology, will be used. The selection process for the new clones and vector design was outsourced to the leading company in the world market and with a recognized reputation Creative Biolabs (USA). Selected antibodies of high specificity against BCMA will give rise to the scFv fragment that will be incorporated into the lentiviral vector with a co-stimulatory molecule (41BB) and a security and labeling gene (EGFR), for later expression in T lymphocytes. Then, the steps mentioned will be performed: 1) production of lentiviral particles and transduction of Jukart cell line; 2) collection of peripheral blood mononuclear cells; 3) lentiviral transduction of T lymphocytes; 4) quality control of the generation of T-CAR cells; 5) immunophenotypic characterization; 6) in vitro cytotoxicity assay and; 7) in vivo cytotoxicity assay with xenotransplant model mice. Memory T cells (central memory or stem central memory) will be selected for genetic modification and evaluation of the cytotoxicity profile and in vivo persistence. At the end of this project, we hope to characterize and validate new lentiviral vectors for the genetic modification of T cells that can express anti-BCMA and, therefore, eradicate Multiple Myeloma plasma cells. Then, it is expected that such a project will support the use of new vectors in the manufacture of anti-BCMA CAR T cells in accordance with standard of Good Manufacturing Practices (GMP) and compatible for use in a phase I clinical trial in humans. (AU)