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Contextual Reprogramming of CAR-T Cells for Treatment of HER2+ Cancers

34 Pages Posted: 14 Aug 2020 Publication Status: Review Complete

See all articles by Zhifen Yang

Zhifen Yang

Refuge Biotechnologies Inc. Menlo Park

Lingyu Li

Refuge Biotechnologies Inc. Menlo Park

Ahu Turkoz

Refuge Biotechnologies Inc. Menlo Park

Pohan Chen

Refuge Biotechnologies Inc. Menlo Park

Rona Harari-Steinfeld

Refuge Biotechnologies Inc. Menlo Park

Alper Kearney

Refuge Biotechnologies Inc. Menlo Park

Dharmesh Patel

Refuge Biotechnologies Inc. Menlo Park

Vitaly Balan

Refuge Biotechnologies Inc. Menlo Park

Maggie Bobbin

Refuge Biotechnologies Inc. Menlo Park

Hana Choi

Refuge Biotechnologies Inc. Menlo Park

Ofir Stefanson

Refuge Biotechnologies Inc. Menlo Park

Damla Inel

Refuge Biotechnologies Inc. Menlo Park

Veena Vinod

ESSA Pharma

Alessandra Cesano

ESSA Pharma

Bing Wang

Refuge Biotechnologies Inc. Menlo Park

Kyung-Ho Roh

University of Alabama at Huntsville - Department of Chemical and Materials Engineering

Lei S. Qi

Stanford University - Department of Bioengineering

Francesco M. Marincola

Refuge Biotechnologies Inc. Menlo Park

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Abstract

Adoptive transfer of chimeric antigen receptor (CAR)-engineered T cells combined with checkpoint inhibition may prevent T cell exhaustion and improve clinical outcomes. However, the approach is limited by cumulative costs and toxicities. To overcome this drawback, we created a CAR-T (RB-340-1) that unites in one product the two modalities: a CRISPR interference-(CRISPRi) circuit prevents programmed cell death protein 1 (PD-1) expression upon antigen-encounter. RB-340-1 is engineered to express an anti-human epidermal growth factor receptor 2 (HER2) CAR single chain variable fragment (scFv), with CD28 and CD3ζ co-stimulatory domains linked to the tobacco etch virus (TEV) protease and a single guide RNA (sgRNA) targeting the PD-1 transcription start site (TSS). A second constructs includes linker for activation of T cells (LAT) fused to nuclease-deactivated spCas9 (dCas9)-Kruppel-associated box (KRAB) via a TEV-cleavable sequence (TCS). Upon antigen encounter, the LAT-dCas9-KRAB (LdCK) complex is cleaved by TEV allowing targeting of dCas9-KRAB to the PD-1 gene TSS. The conditional, non-gene editing and reversible suppression promotes CAR-T cells resilience to checkpoint inhibition, and their persistence and effectiveness against HER2-expressing cancer xenografts.

Suggested Citation

Yang, Zhifen and Li, Lingyu and Turkoz, Ahu and Chen, Pohan and Harari-Steinfeld, Rona and Kearney, Alper and Patel, Dharmesh and Balan, Vitaly and Bobbin, Maggie and Choi, Hana and Stefanson, Ofir and Inel, Damla and Vinod, Veena and Cesano, Alessandra and Wang, Bing and Roh, Kyung-Ho and Qi, Lei S. and Marincola, Francesco M., Contextual Reprogramming of CAR-T Cells for Treatment of HER2+ Cancers. Available at SSRN: https://ssrn.com/abstract=3656606 or http://dx.doi.org/10.2139/ssrn.3656606
This version of the paper has not been formally peer reviewed.

Zhifen Yang

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Lingyu Li

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Ahu Turkoz

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Pohan Chen

Refuge Biotechnologies Inc. Menlo Park

United States

Rona Harari-Steinfeld

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Alper Kearney

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Dharmesh Patel

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Vitaly Balan

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Maggie Bobbin

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Hana Choi

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Ofir Stefanson

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Damla Inel

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Veena Vinod

ESSA Pharma ( email )

Alessandra Cesano

ESSA Pharma ( email )

Bing Wang

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

Kyung-Ho Roh

University of Alabama at Huntsville - Department of Chemical and Materials Engineering ( email )

Huntsville, AL 35899
United States

Lei S. Qi

Stanford University - Department of Bioengineering

Francesco M. Marincola (Contact Author)

Refuge Biotechnologies Inc. Menlo Park ( email )

United States

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