Modification of T cells with anti-CD19 chimeric antigen receptor

• Grant number: 16/08374-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2016
• End date: July 31, 2018
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Virginia Picanço e Castro
• Grantee: Pablo Diego Moço
• Host Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
• Associated research grant: 13/08135-2 - CTC - Center for Cell-Based Therapy, AP.CEPID

Abstract

Immunotherapy with modified T cells to express chimeric antigen receptor (CAR) has been proven effective in remission of acute lymphoblastic leukemia (ALL) in children and young adults. The CAR possess an extracellular domain derived from antibody variable regions, and intracellular domains of T cell costimulation. CD19 protein has been shown to be an ideal target for therapies with CAR because it is present only on the surface of B cells and dendritic follicular cells besides having high expression in virtually all types of leukemias. This paper proposes the production of anti-CD19 CAR-T cells, with 4-1BB as costilulatory intracellular domain. Initially the validation of the production will be carried out by using T cell lines Jurkat or NK-92, which will be transduced with lentiviral vectors produced in HEK293T. The expression of CAR will be verified by flow cytometry and the cytotoxic activity of the modified lines will be evaluated in co-culture with CD19+ B cell lines Raji and Namalwa. Once the production is validated in cell lines, we will begin the modification of peripheral blood T cells isolated from healthy individuals. The isolation of T cells will be made with magnetic beads coupled with anti-CD3 antibodies and the activation of these cells will occur in culture with interleukin IL-2 beads conjugated with anti-CD3 and anti-CD28. The activated T cells will be transduced with the lentiviral vectors. The expression of CAR and activity against B cell lines will be evaluated as the validation. In addition to these assessments, a cytotoxicity assay will be conducted, in which the CAR-T cells will be incubated with peripheral blood mononuclear cells.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

PICANCO-CASTRO, VIRGINIA; MOCO, PABLO DIEGO; MIZUKAMI, AMANDA; VAZ, LETICIA DELFINI; FERNANDES PEREIRA, MARCELO DE SOUZA; SILVESTRE, RENATA NACASAKI; COTTAS DE AZEVEDO, JULIA TEIXEIRA; BOMFIM, ALINE DE SOUSA; DE ABREU NETO, MARIO SOARES; RIBEIRO MALMEGRIM, KELEN CRISTINA; et al. Establishment of a simple and efficient platform for car-t cell generation and expansion: from lentiviral production to in vivo studies. Hematology, Transfusion and Cell Therapy, v. 42, n. 2, p. 150-158, APR-JUN 2020. (17/09491-8, 16/08374-5, 13/08135-2, 12/23228-4)

MARTINS FERNANDES PAES, BARBARA CRISTINA; MOCO, PABLO DIEGO; PEREIRA, CRISTIANO GONCALVES; PORTO, GECIANE SILVEIRA; DE SOUSA RUSSO, ELISA MARIA; JUNQUEIRA REIS, LUIZA CUNHA; COVAS, DIMAS TADEU; PICANCO-CASTRO, VIRGINIA. Ten years of iPSC: clinical potential and advances in vitro hematopoietic differentiation. CELL BIOLOGY AND TOXICOLOGY, v. 33, n. 3, p. 233-250, MAY 2017. (16/08373-9, 13/08135-2, 16/08374-5, 14/22500-8)

PICANCO-CASTRO, VIRGINIA; MOCO, PABLO DIEGO; MIZUKAMI, AMANDA; VAZ, LETICIA DELFINI; FERNANDES PEREIRA, MARCELO DE SOUZA; SILVESTRE, RENATA NACASAKI; COTTAS DE AZEVEDO, JULIA TEIXEIRA; BOMFIM, ALINE DE SOUSA; DE ABREU NETO, MARIO SOARES; RIBEIRO MALMEGRIM, KELEN CRISTINA; et al. Establishment of a simple and efficient platform for car-t cell generation and expansion: from lentiviral production to in vivo studies. Hematology, Transfusion and Cell Therapy, v. 42, n. 2, p. 9-pg., 2020-04-01. (12/23228-4, 17/09491-8, 16/08374-5, 13/08135-2)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

MOÇO, Pablo Diego. Establishment of a platform for the production of lentiviral vectors for the modification of anti-CD19 CAR-T cells. 2018. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) Ribeirão Preto.