The Connell and O'Reilly Families Cell Manipulation Core Facility (CMCF) assists DF/HCC members in developing new cell-based therapies for patients with cancer who are enrolled in novel therapeutic clinical research protocols.
Key Services
Core services include processing of hematopoietic stem cells for autologous or allogeneic transplantation, genetic modification of hematopoietic stem cells, generation of tumor vaccines, and manufacture of genetically modified immune cells for adoptive cellular therapy. The core also collects and stores follow-up blood and bone marrow samples from subjects treated on cell therapy protocols, thus providing an additional service to clinical investigators, ensuring that all research objectives of clinical protocols are met.
Pricing varies; contact the Core for details.
Key services of the Core include:
Production of cellular products for patients enrolled on clinical research protocols
Hematopoietic stem cells
Hematopoietic stem cell graft engineering
Genetically modified stem cells
Genetically modified T cells
Natural Killer cells
Regulatory T cells
Tumor cell vaccines
Dendritic cell vaccines
Induced pluripotent stem cells (iPSC) and iPSC-derived cells
Limbal stem cells
Mesenchymal stromal cells
Cell therapy process development, consultation, and support
Develop manufacturing SOPs
Validate processes and SOPs
Build and activate templates for orders to manufacture, collect, process, and release cellular products to patients (Biotherapy Order Entry, BOE)
Clinical trial development consultation for cellular therapeutic studies
Pre-clinical development and trial design
DF/HCC and FDA Review
Data management
Quality control
Internal and external audits
Reports and publications
Regulatory (internal and external) consultation and support
Assist with pre-IND FDA meetings
Assist with Chemistry, Manufacturing, and Controls (CMC) section of -IND preparation and patient consents
Assist PI with amendments and annual reports
Assist with IRB protocol submission
Review and sign-off on all protocols
Audit all IND/IDE manufacturing for compliance with IND/IDE and protocol
Prepare for FDA and Sponsor audits
Quality control (assays and testing)
Quality assurance
Regulatory (FDA) and accreditation (FACT & JC) compliance monitoring
Audit and review procedures, processes, and test results
Assist with labeling requirements
Monitor product non-conformances
Cellular product manufacturing, characterization, storage, and distribution
Pre- and post-treatment processing and banking of patient samples
Major Equipment
Miltenyi Biotec CliniMACS cell selection device
Miltenyi Biotec CliniMACS Prodigy cell selection and culturing device
BD Beckman Coulter Navios flow cytometer
Sysmex XE-5000 hematology analyzer
BacT/ALERT sterility testing system
ThermoFisher ABI 7500 Fast PCR system w/AccuSEQ
REES Scientific environmental monitoring system
Highlighted projects
The CMCF currently supports more than 40 diverse active clinical research protocols, with a focus on translational research in the immune therapy of cancer. Many of these clinical trials and therapies could not have been carried out without the cell manufacturing and regulatory support provided by the Core.
Notable examples of therapies and protocols CMCF currently supported are shown below:
GENETIC MODIFICATION OF HEMATOPOIETIC STEM CELL GRAFTS
Project Title: Gene transfer for SCID-X1 Principal Investigator: Jennifer Whangbo, MD , PhD (BCH) Sponsor: David A. Williams, MD (BCH) Coordinating Center: Boston Children's Hospital Project Description: A multi-institutional phase I/II clinical trial of somatic gene therapy for patients with SCID-X1 using bone marrow derived CD34+ cells transduced ex-vivo with a novel gibbon ape leukemia (GALV)-virus, pseudotyped gammaretroviral vector encoding the human common cytokine receptor gamma chain (γc). This clinical trial is performed in collaboration with two other sites in the United States and two sites in Europe. The CMCF is responsible for in vitro processing of patient's bone marrow, which included selection of CD34+ stem cells and transduction with retroviral vector.
Project Title: Gene therapy for patients with Sickle Cell Disease (SCD) Principal Investigator: Erica Esrick, MD (BCH) Sponsor:David A. Williams, MD (BCH) Coordinating Center: Boston Children’s Hospital Project Description: A Multi-Center, Phase 2 Gene Transfer Study Inducing Fetal Hemoglobin in Sickle Cell Disease. Following successful demonstration of safety and early clinical efficacy in a Phase I trial in a clinical trial at BCH, this clinical trial will examine the efficacy of autologous transplantation of stem cells genetically engineered to inhibit expression of BCL11A and enhance expression of fetal hemoglobin (HbF). Expression of HbF effectively prevents sickling of red blood cells and as shown in the phase I trial, prevents hemolysis, allows normalization of red cell counts without transfusion and alleviates manifestations of severe sickle cell disease. The CMCF is one of 2 cell manufacturing sites for this national multicenter clinical trial supported by NHLBI and CIRM.
CHIMERIC ANTIGEN RECEPTOR T CELLS.
Autologous T cells genetically engineered to express receptors targeting tumor cells have demonstrated remarkable efficacy in adult and pediatric patients with refractory B cell malignancies. Several CD19-CAR T cell products have been FDA approved since 2017. The CMCF is currently supporting several Phase I/II CAR T cell protocols with innovative cellular products. CAR T cells are manufactured using the Miltenyi CliniMACS Prodigy instrument.
Project Title: A Phase I clinical trial with CAR-37 T cells for patients with relapsed or refractory CD37+ hematologic malignancies (DF/HCC 19-087) Principal Investigator: Matt Frigault, MD (MGH) Sponsor: Marcela Maus, MD, PhD (MGH) Project Description: This trial is based on preclinical studies in Dr. Maus’ lab identifying CD37 as a novel surface antigen expressed on most B cell lymphomas and some T cell lymphomas. This is the first-in-human CAR T cell therapy targeting CD37. Patients with T cell lymphoma and patients who relapse after CD19 CAR T cells therapy are eligible for this clinical trial if tumor cells express CD37.
Project Title: Phase 1 cell therapy for the treatment of patients with relapsed and refractory multiple myeloma (DF/HCC 19-253) Principal Investigators: Matt Frigault, MD (MGH) Jacalyn Rosenblatt MD (BIDMC) Sponsor: Arcellx. Project Description: Unlike other CAR T cell vectors that utilize scFv antibodies to mediate specificity, this construct employs a proprietary synthetic receptor specific for BCMA.
Project Title: Phase I study of BCMA-directed CAR-T cells in adult patients with relapsed and/or refractory multiple myeloma (DF/HCC 19-785) Principal Investigators: Adam Sperling MD (DFCI), Nikhil Munshi,MD (DFCI) Sponsor: Novartis. Project Description: This clinical trial employs a unique manufacturing process to generate highly functional BCMA-CAR T cells. The manufacturing process was developed in collaboration with Novartis and if successful will be incorporated in other Novartis-sponsored protocols for deployment in future trials.
Project Title: A Phase I study to investigate TEG002 infusion in relapsed/refractory multiple myeloma patients (DF/HCC 20-126) Principal Investigators: Adam Sperling MD (DFCI), Nikhil Munshi,MD (DFCI) Sponsor: Gadeta. Project Description: This clinical trial utilizes a unique TCRgd eceptor (TEG002) whose expression is induced on conventional TCRab T cells. TEG002 cells are specific for CD227J, a unique epitope widely expressed on hematologic and non-hematologic tumors. The specificity of TEG002 is not HLA-restricted. Expression of CD277J on tumor cells is increased in the presence of pamidronate or zoledronate which further enhances the tumor cell specificity of these genetically engineered effecter cells.
CYTOKINE-INDUCED MEMORY-LIKE NK CELLS (CIML-NK).
Previous studies have demonstrated that brief in-vitro activation of resting peripheral blood NK cells with IL-12, IL-15, and IL-18 results in the differentiation of Cytokine-Induced Memory-Like (CIML) NK cells with potent anti-leukemia activity. The CMCF is supporting several clinical trials manufacturing highly purified CIML NK cells that are infused immediately after in vitro activation, without cryopreservation.
Project Title: A phase 1 trial of CIML NK cell infusion for myeloid disease relapse after haploidentical hematopoietic cell transplantation (DF/HCC 19-265) Principal Investigator: Rizwan Romee,MD (DFCI) Sponsor: Rizwan Romee,MD (DFCI) Project Description: CIML-NK cells are generated from apheresis products collected from the stem cell donor and will therefore not be recognized as foreign in the transplant recipient. CIML-NK cell infusions are followed by administration of low-dose IL-2 to support expansion and persistence of these cells in vivo.
Project Title: A Phase 1 trial of CTLA-4 inhibition in combination with CIML-NK cell therapy in advanced head & neck cancer (DF/HCC 19-505) Principal Investigator: Glenn Hanna, MD (DFCI) Sponsor: Rizwan Romee,MD (DFCI) Project Description: CIML-NK cells are generated from apheresis products collected from HLA haplotype mismatched related donors. Patients also receive ipilimumab prior to infusion of CIML-NK cells and N-803 IL-15 after infusion to support expansion and persistence of these cells in vivo.
CULTIVATED CORNEAL EPITHELIAL CELLS FOR TRANSPLANTATION IN PATIENTS WITH UNILATERAL LIMBAL STEM CELL DEFICIENCY
Project Title: Cultivated Autologous Limbal Epithelial Cell (CALEC) Transplantation Principal Investigators: Ula Jurkunas, MD (MEEI), Jerome Ritz, MD (DFCI), Allison Ayala MS (Jaeb) Co-Investigators: Reza Dana, MD (MEEI), Jia Yin, MD, PhD (MEEI) Coordinating Centers: Massachusetts Eye and Ear Infirmary, Dana-Farber Cancer Institute and Jaeb Center for Health Research Description of Project: This NIH-NEI-funded study (NCT02592330) with an FDA-approved IND (#16102) involves performing a small limbal biopsy from the healthy eye, which is used to expand limbal stem cells (CALEC cells). After a 2- to 3-week period, CALEC grafts are transplanted onto the eye with limbal stem cell deficiency. This open label, randomized study will enroll 18 patients with unilateral limbal stem cell efficiency; All patients will be followed for 18 months.
The CMCF has developed a GMP-compliant process for reprogramming T cells into iPSC. We chose T cells as our starting material since these cells all harbor genetically rearranged TCR genes that can be used as molecular markers of clonality. The specific TCR gene rearrangement of each iPSC line also serves as a genetic barcode that can be used to track all differentiated cellular products derived from these iPSC. This work has been a collaborative effort with investigators who have developed methods for differentiating iPSC into pancreatic islet cells and dopaminergic neurons and the CMCF will be supporting the manufacture of these differentiated cells for autologous transplantation in future clinical trials in patients with diabetes and Parkinson’s disease. With this GMP-compliant platform in place for generating iPSC, we are now also able to develop protocols for generating differentiated NK cells or T cells for cancer therapy. Future studies will incorporate genetic modifications of iPSC lines to enhance efficacy of derived immune effector cells. Genetic modifications to reduce the immunogenicity of these cells will support the development of large iPSC banks that can be used to generate allogeneic NK or T cells for adoptive therapy in patients with cancer.
Publication Acknowledgement
If research supported by this core facility results in publication, please acknowledge this support by including the following in your publication(s): "We thank Dana-Farber/Harvard Cancer Center in Boston, MA, for the use of the Cell Manipulation Core, which provided __________ service. Dana-Farber/Harvard Cancer Center is supported in part by an NCI Cancer Center Support Grant # NIH 5 P30 CA06516."
