Manufacturing and preclinical validation of CAR T cells targeting ICAM-1 for advanced thyroid cancer therapy

• Published in: Scientific reports Vol. 9; no. 1; p. 10634
• Main Authors: Vedvyas, Yogindra, McCloskey, Jaclyn E., Yang, Yanping, Min, Irene M., Fahey, Thomas J., Zarnegar, Rasa, Hsu, Yen-Michael S., Hsu, Jing-Mei, Van Besien, Koen, Gaudet, Ian, Law, Ping, Kim, Nak Joon, Hofe, Eric von, Jin, Moonsoo M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 23-07-2019; Nature Publishing Group
• Subjects: 13; 38; 42; 59/5; 59/78; 631/250/251; 631/67/1059/2325; 64/60; 82; 82/51; Affinity; Animals; Antigens; Antitumor agents; Apheresis; Cancer therapies; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; Cell culture; Cell Survival - immunology; Cross Reactions; Cross-reactivity; Cryopreservation; Cytotoxicity; HEK293 Cells; HeLa Cells; Humanities and Social Sciences; Humans; Immunotherapy, Adoptive - methods; Intercellular adhesion molecule 1; Intercellular Adhesion Molecule-1 - immunology; Intercellular Adhesion Molecule-1 - metabolism; Leukocytes; Lymphocytes; Lymphocytes T; Malignancy; Medical prognosis; Mice; Mice, Inbred BALB C; multidisciplinary; Patients; Peripheral blood; Receptors, Chimeric Antigen - immunology; Science; Science (multidisciplinary); Thawing; Thyroid; Thyroid cancer; Thyroid carcinoma; Thyroid Neoplasms - therapy; Transduction, Genetic; Tumors; Xenograft Model Antitumor Assays - methods
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-46938-7

While the majority of thyroid cancer patients are easily treatable, those with anaplastic or poorly differentiated recurrent thyroid carcinomas have a very poor prognosis with a median survival of less than a year. Previously, we have shown a significant correlation between ICAM-1 overexpression and malignancy in thyroid cancer, and have pioneered the use of ICAM-1 targeted CAR T cells as a novel treatment modality. For clinical translation of this novel modality, we designed CAR T cells possessing micromolar rather than nanomolar affinity to ICAM-1 to avoid cytotoxicity in normal cells with basal levels of ICAM-1 expression. Herein, we report the automated process of CAR T cell manufacturing with CliniMACS Prodigy (Miltenyi Biotec) using cryopreserved peripheral blood leukocytes from apheresis collections. Using Prodigy, thawed leukopak cells were enriched for CD4
+
and CD8
+
T cells, subjected to double transduction using lentiviral vector, and expanded in culture for a total of 10 days with a final yield of 2–4 × 10
9
cells. The resulting CAR T cells were formulated for cryopreservation to be used directly for infusion into patients after thawing with no further processing. We examined cross-reactivity of CAR T cells toward both human and murine ICAM-1 and ICAM-1 expression in human and mouse tissues to demonstrate that both efficacy and on-target, off-tumor toxicity can be studied in our preclinical model. Selective anti-tumor activity in the absence of toxicity provides proof-of-concept that micromolar affinity tuned CAR T cells can be used to target tumors expressing high levels of antigen while avoiding normal tissues expressing basal levels of the same antigen. These studies support the initiation of a phase I study to evaluate the safety and potential efficacy of micromolar affinity tuned CAR T cells against newly diagnosed anaplastic and refractory or recurrent thyroid cancers.