Manufacturing development and clinical production of NKG2D chimeric antigen receptor-expressing T cells for autologous adoptive cell therapy
Joana M Murad, Susanne H Baumeister, Lillian Werner, Heather Daley, Hélène Trébéden-Negre, Jake Reder, Charles L Sentman, David Gilham, Frederic Lehmann, Sarah Snykers, Marie-Louise Sentman, Terri Wade, Adam Schmucker, Michael W Fanger, Glenn Dranoff, Jerome Ritz, Sarah Nikiforow, Joana M Murad, Susanne H Baumeister, Lillian Werner, Heather Daley, Hélène Trébéden-Negre, Jake Reder, Charles L Sentman, David Gilham, Frederic Lehmann, Sarah Snykers, Marie-Louise Sentman, Terri Wade, Adam Schmucker, Michael W Fanger, Glenn Dranoff, Jerome Ritz, Sarah Nikiforow
Abstract
Background aims: Adoptive cell therapy employing natural killer group 2D (NKG2D) chimeric antigen receptor (CAR)-modified T cells has demonstrated preclinical efficacy in several model systems, including hematological and solid tumors. We present comprehensive data on manufacturing development and clinical production of autologous NKG2D CAR T cells for treatment of acute myeloid leukemia and multiple myeloma (ClinicalTrials.gov Identifier: NCT02203825). An NKG2D CAR was generated by fusing native full-length human NKG2D to the human CD3ζ cytoplasmic signaling domain. NKG2D naturally associates with native costimulatory molecule DAP10, effectively generating a second-generation CAR against multiple ligands upregulated during malignant transformation including MIC-A, MIC-B and the UL-16 binding proteins.
Methods: CAR T cells were infused fresh after a 9-day process wherein OKT3-activated T cells were genetically modified with replication-defective gamma-retroviral vector and expanded ex vivo for 5 days with recombinant human interleukin-2.
Results: Despite sizable interpatient variation in originally collected cells, release criteria, including T-cell expansion and purity (median 98%), T-cell transduction (median 66% CD8+ T cells), and functional activity against NKG2D ligand-positive cells, were met for 100% of healthy donors and patients enrolled and collected. There was minimal carryover of non-T cells, particularly malignant cells; both effector memory and central memory cells were generated, and inflammatory cytokines such as granulocyte colony-stimulating factor, RANTES, interferon-γ and tumor necrosis factor-α were selectively up-regulated.
Conclusions: The process resulted in production of required cell doses for the first-in-human phase I NKG2D CAR T clinical trial and provides a robust, flexible base for further optimization of NKG2D CAR T-cell manufacturing.
Keywords: CAR T; Good manufacturing practice cell therapy; NKG2D; acute myeloid leukemia; chimeric antigen receptor; gamma retrovirus; multiple myeloma.
Conflict of interest statement
Disclosures:
C.L.S. has patents and financial interests in NK receptor–based CAR therapies. C.L.S. is a scientific founder for Celdara Medical, a consultant, and receives research support from Celdara Medical. These conflicts are managed under the policies of Dartmouth College. M.-L.S. has an immediate family member with financial interests in NK receptor–based CAR therapies. J.M.M., J.R., M.W.F., T.W., and A.S. are employed by Celdara Medical, which has a material financial interest in NK receptor–based CAR intellectual property assigned to the Trustees of Dartmouth College. DEG, SS and FLE are employees of Celyad S.A, which is the clinical trial sponsor. G.D. is currently an employee of Novartis, which has material financial interests in other CAR T cell therapies. S.B., S.N. received salary support through an SBIR grant awarded to Celdara Medical. The other authors have no financial conflicts of interest.
Copyright © 2018 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
Figures
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Figure 4
Transduction efficiency, phenotype and functional…
Figure 4
Transduction efficiency, phenotype and functional activity of AML and MM-derived NKG2D CAR-transduced T-cell…
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- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't
- Cell Line, Tumor
- Cell Proliferation
- Clinical Trials as Topic
- Cytokines / metabolism
- Humans
- Immunotherapy, Adoptive*
- Ligands
- NK Cell Lectin-Like Receptor Subfamily K / metabolism*
- Phenotype
- Receptors, Antigen, T-Cell / immunology
- Receptors, Chimeric Antigen / metabolism*
- T-Lymphocytes / cytology
- T-Lymphocytes / immunology*
- Transplantation, Autologous
- Cytokines
- Ligands
- NK Cell Lectin-Like Receptor Subfamily K
- Receptors, Antigen, T-Cell
- Receptors, Chimeric Antigen
- ClinicalTrials.gov/NCT02203825
- Full Text Sources
- Other Literature Sources
- Medical
- Research Materials
- Miscellaneous
![Figure 4](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/6127861/bin/nihms979356f4.jpg)
Source: PubMed