Manufacturing Validation of Biologically Functional T Cells ... : Journal of Immunotherapy (original) (raw)

Clinical Studies

Manufacturing Validation of Biologically Functional T Cells Targeted to CD19 Antigen for Autologous Adoptive Cell Therapy

Hollyman, Daniel*; Stefanski, Jolanta*; Przybylowski, Mark*; Bartido, Shirley*; Borquez-Ojeda, Oriana*; Taylor, Clare*; Yeh, Raymond§; Capacio, Vanessa*; Olszewska, Malgorzata*; Hosey, James*; Sadelain, Michel* † ‡ §; Brentjens, Renier J.† §; Rivière, Isabelle* † ‡ §

*Gene Transfer and Somatic Cell Engineering Facility

†Center for Cell Engineering

‡Molecular Pharmacology and Chemistry Program

§Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY

Supported by The Alliance for Cancer Gene Therapy P30 CA-008748, PO1 CA-059350, PO1 CA-023766, P50 CA086438, Damon Runyon Clinical Investigator Award (R.J.B.), the Annual Terry Fox Run for Cancer Research (New York, NY) organized by the Canada Club of New York, the Geoffrey Beene Cancer Foundation, Golfers Against Cancer, and by Mr William H. Goodwin and Mrs Alice Goodwin of the Commonwealth Cancer Foundation for Research and the Experimental Therapeutics Center of MSKCC.

Financial disclosure: All authors have declared there are no financial conflicts of interest in regard to this work.

Reprints: Isabelle Rivière, Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 182, New York, NY 10021 (e-mail: [email protected]).

Received for publication September 9, 2008; accepted October 26, 2008

Abstract

On the basis of promising preclinical data demonstrating the eradication of systemic B-cell malignancies by CD19-targeted T lymphocytes in vivo in severe combined immunodeficient-beige mouse models, we are launching phase I clinical trials in patients with chronic lymphocytic leukemia (CLL) and acute lymphoblastic leukemia. We present here the validation of the bioprocess which we developed for the production and expansion of clinical grade autologous T cells derived from patients with CLL. We demonstrate that T cells genetically modified with a replication-defective gammaretroviral vector derived from the Moloney murine leukemia virus encoding a chimeric antigen receptor (CAR) targeted to CD19 (1928z) can be expanded with Dynabeads CD3/CD28. This bioprocess allows us to generate clinical doses of 1928z+ T cells in approximately 2 to 3 weeks in a large-scale semiclosed culture system using the Wave Bioreactor. These 1928z+ T cells remain biologically functional not only in vitro but also in severe combined immunodeficient-beige mice bearing disseminated tumors. The validation requirements in terms of T-cell expansion, T-cell transduction with the 1928z CAR, biologic activity, quality control testing, and release criteria were met for all 4 validation runs using apheresis products from patients with CLL. Additionally, after expansion of the T cells, the diversity of the skewed Vβ T-cell receptor repertoire was significantly restored. This validated process will be used in phase I clinical trials in patients with chemorefractory CLL and in patients with relapsed acute lymphoblastic leukemia. It can also be adapted for other clinical trials involving the expansion and transduction of patient or donor T cells using any CAR or T-cell receptor.