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Cell Manipulation Core Facility

The Cell Manipulation Core Facility (CMCF) produces safe and effective cellular components for subjects enrolled in novel therapeutic clinical research protocols. The CMCF currently supports more than 40 active clinical research protocols, and core users work in diverse areas, including hematopoietic stem cell transplantation, cancer vaccines, and adoptive cellular therapy of cancer. 

Linda Kelley, PhD, was recently recruited from the University of Utah, where she was the Director of their Cell Therapy Facility, to serve as the Director of the CMCF.

Overview

In the past five years, the DF/HCC Cancer Immunology Program has made a major effort to support translational research in the immune therapy of cancer. This effort resulted in the initiation of a large number of clinical trials in hematopoietic stem cell transplantation focusing on modulation of immune reconstitution to either enhance graft versus leukemia (GVL) or suppress graft versus host disease (GVHD) after transplant. A large number of studies also examined the safety and efficacy of autologous tumor vaccines in a variety of different diseases. In many cases, vaccine therapy was administered in combination with other treatments to enhance and maintain the immune response generated by the tumor vaccine. The CMCF has played a major role in many of these trials, which could not have been carried out without the cell manufacturing and regulatory support provided by the Core.

The clinical research protocols that are currently supported by the CMCF represent a variety of studies developing new therapies for subjects with different malignant diseases. The vast majority of hematopoietic stem cell transplantation protocols target subjects with hematologic malignancies while the tumor vaccine studies also target subjects with solid tumors. In each instance, these clinical trials represent a collaborative effort between clinical and laboratory investigators. Often these investigators are based at different institutions within DF/HCC and these studies represent collaborations between the Cancer Immunology Program and different Clinical Science Programs. The CMCF produces manipulated hematopoietic progenitor cell (HPC) components for adult patients treated by the joint Dana-Farber/Brigham and Women’s Cancer Center (DF/BWCC) hematopoietic stem cell transplant program and for pediatric patients treated by the Dana-Farber/Children’s Hospital Cancer Care (DF/CHCC) bone marrow transplant program. Manipulated stem cells for specific protocols are also provided for patients at Massachusetts General Hospital (MGH) and Beth Israel Deaconess Medical Center (BIDMC). Similarly, cellular components prepared using ex vivo culture and/or viral vector transduction for clinical trials in immunotherapy currently involve patients treated at DF/BWCC, DF/CHCC and MGH.

As new clinical trials are being developed, the CMCF also works with Cancer Center members to develop and optimize manufacturing procedures for their clinical trials and to support FDA IND applications. To ensure patient safety and comply with FDA regulations, cellular products that are processed under investigational methods must meet specific safety requirements, quality control tests and release criteria prior to human use. Assays to accurately characterize and assess the potency of specific cellular products are developed for different protocols and become important measures of manufacturing efficacy. In the last two years, the CMCF has developed its own Quality Control lab with trained staff and qualified instruments to support the development and implementation of these assays. Flow cytometry-based product characterization and functional assays as well as testing for sterility, endotoxin and environmental monitoring are carried out in the QC lab. This has markedly reduced the cost and turn around time for results reporting. Procedures for transporting clinical products between the CMCF and all of the affiliated clinical sites within DF/HCC have also been validated to allow the CMCF to receive, dispense, and support clinical studies at all DF/HCC clinical sites.

Protocols involving the manipulation of hematopoietic progenitor cells often require assessment of engraftment and immunologic reconstitution. To support these important research objectives, CMCF also collects and stores follow-up blood and bone marrow samples collected from subjects treated on the protocols for which the facility produces components. In this way, the CMCF provides an additional service to clinical investigators and ensures that all of the research objectives of the clinical protocol are met. Although this service goes far beyond the services normally provided by cell manufacturing laboratories, the CMCF considers this an essential component of the core’s work in support of clinical research within DF/HCC.

Project Examples

DF/HCC Protocol 04-023 (BB-IND 7248): GM-CSF secreting leukemia cell vaccinations after allogeneic non-myeloablative peripheral blood stem cell transplantation in patients with advanced myelodysplastic syndrome or refractory acute or advanced chronic myeloid leukemia. PIs: V. HoDFCI, R. SoifferDFCI and G. DranoffDFCI. Cancer Center Programs: Leukemia and Cancer Immunology
IND Sponsor: G. DranoffDFCI. Funding Source: NIH-NIAID.

The early post-transplant period after allogeneic hematopoietic stem cell transplantation (HSCT) is characterized by severe lymphopenia and intense homeostatic proliferation of donor T, B and NK cells. V. HoDFCI and colleagues hypothesized that administration of autologous tumor vaccines during this period would elicit strong leukemia-specific responses from the donor immune cells. Indeed, pre-clinical experiments in murine model systems demonstrated that GM-CSF-secreting tumor vaccines (GVAX) following T-cell depleted allogeneic HSCT stimulated potent anti-tumor immunity without the exacerbation of GVHD. This study was designed to investigate the safety of leukemia GVAX in patients with refractory MDS, AML or advanced CML when administered early after non-myeloablative allogeneic HSCT. A potential concern was whether vaccination with recipient leukemia cells would increase the frequency and/or severity of GVHD. Cellular vaccines were generated by adenoviral vector-mediated GM-CSF gene transfer into myeloblasts harvested prior to HSCT. After engraftment, between day +30 to +45, patients began a series of vaccinations with lethally irradiated, ad-GM-CSF infected autologous leukemia cells. A total of six vaccinations was planned for each patient. The CMCF processed allogeneic peripheral blood stem cell products and successfully manufactured autologous AML vaccines for 26 subjects enrolled in this study. The study was completed and results were published. A follow-up study using a modified vaccine consisting of autologous AML blasts admixed with K562 cells genetically engineered to secrete GM-CSF is currently accruing subjects. This project resulted in a publication, Ho V, Vanneman M, Kim H, Pasek M, Cutler C, Koreth J, Alyea E, Sarantopoulos S, Antin JH, Ritz J, Canning C, Kutok J, Mihm M, Dranoff G, Soiffer RJ. Biologic activity of irradiated, autologous, GM-CSF secreting leukemia cell vaccines early after allogeneic stem cell transplantation. Proc Natl Acad Sci USA 2009; 106:15825-30.

DF/HCC Protocol 08-274 (BB IND 13721): A Phase I Study of Reduced Intensity, Sequential Double Umbilical Cord Blood Transplantation Using Ex Vivo dmPGE2 Expanded Umbilical Cord Blood Units. Principal Investigators: C. CutlerDFCI, K. BallenMGH and D. AviganBIDMC. Cancer Center Programs: Leukemia and Cancer Immunology. IND Sponsors: L. ZonCHB and Fate Therapeutics, San Diego, CA. Funding Source: NIH-NHLBI.

L. ZonCHB recently discovered that compounds that enhance prostaglandin (PG) E2 synthesis also increase the number of hematopoietic stem cells (HSCs) at early stages of hematopoiesis. A stable derivative of PGE2 (dmPGE2) significantly improved kidney marrow recovery following irradiation injury in the adult zebra fish. In murine models, dmPGE2 caused a 2 to 4-fold amplification of multipotent hematopoietic progenitors. The conserved role for PGE2 in the regulation of vertebrate HSC homeostasis suggests that modulation of the prostaglandin pathway will help HSC expansion for therapeutic purposes. This study proposes to evaluate the safety of dmPGE2 when used ex vivo to enhance engraftment following human unrelated cord blood (UCB) transplants. Adult patients who receive UCB stem cells often experience delayed engraftment and are at increased risk of infections following transplant. As a result, two human cord blood products are often used in adult transplantation to improve hematopoietic reconstitution. In this Phase I study, one of the two cords is treated with dmPGE2 prior to transplantation. The untreated UBC provides a backup source of HSC in the event that treatment with dmPGE2 results in unexpected toxicity to HSC in one product. The degree of bone marrow chimerism in transplant recipients will be analyzed over time in conjunction with granulocyte and platelet engraftment. Immunologic reconstitution is also monitored and compared to previous patients who received similar preparative regimens and untreated double UCB transplants. The CMCF assisted in the IND submission, developed standard operating procedures (SOP) and carried out validations of the manufacturing process. The CMCF has completed processing of dmPGE2 treated cord blood cells for nine patients and the study is ongoing.

Contact
Gerry MacDonald
Cell Manipulation Core Facility
Phone: (617) 632-2251