Enhanced safety and efficacy of protease-regulated CAR-T cell receptors

Louai Labanieh; Robbie G Majzner; Dorota Klysz; Elena Sotillo; Chris J Fisher; José G Vilches-Moure; Kaithlen Zen B Pacheco; Meena Malipatlolla; Peng Xu; Jessica H Hui; Tara Murty; Johanna Theruvath; Nishant Mehta; Sean A Yamada-Hunter; Evan W Weber; Sabine Heitzeneder; Kevin R Parker; Ansuman T Satpathy; Howard Y Chang; Michael Z Lin; Jennifer R Cochran; Crystal L Mackall

doi:10.1016/j.cell.2022.03.041

Enhanced safety and efficacy of protease-regulated CAR-T cell receptors

Cell. 2022 May 12;185(10):1745-1763.e22. doi: 10.1016/j.cell.2022.03.041. Epub 2022 Apr 27.

Authors

Louai Labanieh¹, Robbie G Majzner², Dorota Klysz³, Elena Sotillo³, Chris J Fisher³, José G Vilches-Moure⁴, Kaithlen Zen B Pacheco³, Meena Malipatlolla³, Peng Xu³, Jessica H Hui³, Tara Murty⁵, Johanna Theruvath⁶, Nishant Mehta⁷, Sean A Yamada-Hunter³, Evan W Weber³, Sabine Heitzeneder³, Kevin R Parker⁸, Ansuman T Satpathy⁹, Howard Y Chang¹⁰, Michael Z Lin¹¹, Jennifer R Cochran¹², Crystal L Mackall¹³

Affiliations

¹ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
² Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
³ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
⁴ Department of Comparative Medicine, School of Medicine, Stanford University, Stanford, CA 94305, USA.
⁵ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Medical Scientist Training Program, Stanford University School of Medicine, Stanford, CA 94305, USA; Biophysics Program, Stanford University School of Medicine, Stanford, CA 94305, USA.
⁶ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
⁷ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
⁸ Center for Personal Dynamic Regulomes, Stanford University School of Medicine, Stanford, CA 94305, USA.
⁹ Department of Pathology, Stanford University, Stanford, CA 94305, USA.
¹⁰ Center for Personal Dynamic Regulomes, Stanford University School of Medicine, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
¹¹ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
¹² Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA.
¹³ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: cmackall@stanford.edu.

Abstract

Regulatable CAR platforms could circumvent toxicities associated with CAR-T therapy, but existing systems have shortcomings including leakiness and attenuated activity. Here, we present SNIP CARs, a protease-based platform for regulating CAR activity using an FDA-approved small molecule. Design iterations yielded CAR-T cells that manifest full functional capacity with drug and no leaky activity in the absence of drug. In numerous models, SNIP CAR-T cells were more potent than constitutive CAR-T cells and showed diminished T cell exhaustion and greater stemness. In a ROR1-based CAR lethality model, drug cessation following toxicity onset reversed toxicity, thereby credentialing the platform as a safety switch. In the same model, reduced drug dosing opened a therapeutic window that resulted in tumor eradication in the absence of toxicity. SNIP CARs enable remote tuning of CAR activity, which provides solutions to safety and efficacy barriers that are currently limiting progress in using CAR-T cells to treat solid tumors.

Keywords: CAR-T therapy; T cell differentiation; T cell exhaustion; brain tumors; cellular immunotherapy; chimeric antigen receptors; immune cell engineering; solid cancers; synthetic biology.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

Humans
Immunotherapy, Adoptive / methods
Neoplasms* / drug therapy
Neoplasms* / pathology
Peptide Hydrolases
Receptors, Antigen, T-Cell
Receptors, Chimeric Antigen*
T-Lymphocytes / pathology

Substances

Receptors, Antigen, T-Cell
Receptors, Chimeric Antigen
Peptide Hydrolases

Abstract

Publication types

MeSH terms

Substances

Grants and funding