Research Article
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Published Online: 1 May 2018

Development of Anti-Human Mesothelin-Targeted Chimeric Antigen Receptor Messenger RNA–Transfected Peripheral Blood Lymphocytes for Ovarian Cancer Therapy

Publication: Human Gene Therapy
Volume 29, Issue Number 5

Abstract

CD19-targeted chimeric antigen receptor (CAR) engineered T/natural killer (NK)-cell therapies can result in durable clinical responses in B-cell malignancies. However, CAR-based immunotherapies have been much less successful in solid cancers, in part due to “on-target off-tumor” toxicity related to expression of target tumor antigens on normal tissue. Based on preliminary observations of safety and clinical activity in proof-of-concept clinical trials, tumor antigen-specific messenger RNA (mRNA) CAR transfection into selected, activated, and expanded T/NK cells may permit prospective control of “on-target off-tumor” toxicity. To develop a commercial product for solid tumors, mesothelin was selected as an antigen target based on its association with poor prognosis and overexpression in multiple solid cancers. It was hypothesized that selecting, activating, and expanding cells ex vivo prior to mRNA CAR transfection would not be necessary, thus simplifying the complexity and cost of manufacturing. Now, the development of anti-human mesothelin mRNA CAR transfected peripheral blood lymphocytes (CARMA-hMeso) is reported, demonstrating the manufacture and cryopreservation of multiple cell aliquots for repeat administrations from a single human leukapheresis. A rapid, automated, closed system for cGMP-compliant transfection of mRNA CAR in up to 20 × 109 peripheral blood lymphocytes was developed. Here we show that CARMA-hMeso cells recognize and lyse tumor cells in a mesothelin-specific manner. Expression of CAR was detectable over approximately 7 days in vitro, with a progressive decline of CAR expression that appears to correlate with in vitro cell expansion. In a murine ovarian cancer model, a single intraperitoneal injection of CARMA-hMeso resulted in the dose-dependent inhibition of tumor growth and improved survival of mice. Furthermore, repeat weekly intraperitoneal administrations of the optimal CARMA-hMeso dose further prolonged disease control and survival. No significant off-target toxicities were observed. These data support further investigation of CARMA-hMeso as a potential treatment for ovarian cancer and other mesothelin-expressing cancers.

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Information & Authors

Information

Published In

cover image Human Gene Therapy
Human Gene Therapy
Volume 29Issue Number 5May 2018
Pages: 614 - 625
PubMed: 29334771

History

Published in print: May 2018
Published online: 1 May 2018
Published ahead of print: 2 April 2018
Published ahead of production: 15 January 2018
Accepted: 19 December 2017
Received: 16 May 2017

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Authors

Affiliations

Chien-Fu Hung
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland
Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland
Xuequn Xu
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland
Linhong Li
MaxCyte, Inc., Gaithersburg, Maryland
Ying Ma
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland
Qiu Jin
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland
Angelia Viley
MaxCyte, Inc., Gaithersburg, Maryland
Cornell Allen
MaxCyte, Inc., Gaithersburg, Maryland
Pachai Natarajan
MaxCyte, Inc., Gaithersburg, Maryland
Rama Shivakumar
MaxCyte, Inc., Gaithersburg, Maryland
Madhusudan V. Peshwa
MaxCyte, Inc., Gaithersburg, Maryland
Leisha A. Emens* [email protected]
Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland
Johns Hopkins Bloomberg–Kimmel Institute for Cancer Immunotherapy, Baltimore, Maryland.

Notes

These authors contributed equally to this work.
*
Correspondence: Dr. Leisha A. Emens, Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 409, Cancer Research Building 1, Baltimore, MD 21231. E-mail: [email protected]

Author Disclosure

These studies were funded by MaxCyte, Inc. C.F.H. and L.A.E. received financial and material support from MaxCyte, Inc. L.L., A.V., C.A., P.N., R.S., and M.V.P. are employees of and have personal financial interests in MaxCyte, Inc.

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