A Non-integrating Lentiviral Approach Overcomes Cas9-Induced Immune Rejection to Establish an Immunocompetent Metastatic Renal Cancer Model

Junhui Hu; Shiruyeh Schokrpur; Maani Archang; Kip Hermann; Allison C. Sharrow; Prateek Khanna; Jesse Novak; Sabina Signoretti; Rupal S. Bhatt; Beatrice S. Knudsen; Hua Xu; Lily Wu

Molecular Therapy: Methods & Clinical Development (Jun 2018)

A Non-integrating Lentiviral Approach Overcomes Cas9-Induced Immune Rejection to Establish an Immunocompetent Metastatic Renal Cancer Model

Junhui Hu,
Shiruyeh Schokrpur,
Maani Archang,
Kip Hermann,
Allison C. Sharrow,
Prateek Khanna,
Jesse Novak,
Sabina Signoretti,
Rupal S. Bhatt,
Beatrice S. Knudsen,
Hua Xu,
Lily Wu

Affiliations

Junhui Hu: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; Department of Paediatric Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Shiruyeh Schokrpur: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
Maani Archang: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
Kip Hermann: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
Allison C. Sharrow: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
Prateek Khanna: Department of Medicine, Beth Israel Deacones Medical Center, Boston, MA 02215, USA; Kidney Cancer Program, Dana-Farber Harvard Cancer Center, Boston, MA 02215, USA
Jesse Novak: Kidney Cancer Program, Dana-Farber Harvard Cancer Center, Boston, MA 02215, USA; Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02215, USA; Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215, USA
Sabina Signoretti: Kidney Cancer Program, Dana-Farber Harvard Cancer Center, Boston, MA 02215, USA; Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02215, USA; Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215, USA
Rupal S. Bhatt: Department of Medicine, Beth Israel Deacones Medical Center, Boston, MA 02215, USA; Kidney Cancer Program, Dana-Farber Harvard Cancer Center, Boston, MA 02215, USA
Beatrice S. Knudsen: Department of Pathology, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA
Hua Xu: Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Lily Wu: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Urology, Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Corresponding author: Lily Wu, MD, PhD, Departments of Molecular & Medical Pharmacology and Urology, 33-118 CHS, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1735, USA.

Journal volume & issue: Vol. 9
pp. 203 – 210

Abstract

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The CRISPR-based technology has revolutionized genome editing in recent years. This technique allows for gene knockout and evaluation of function in cell lines in a manner that is far easier and more accessible than anything previously available. Unfortunately, the ability to extend these studies to in vivo syngeneic murine cell line implantation is limited by an immune response against cells transduced to stably express Cas9. In this study, we demonstrate that a non-integrating lentiviral vector approach can overcome this immune rejection and allow for the growth of transduced cells in an immunocompetent host. This technique enables the establishment of a von Hippel-Lindau (VHL) gene knockout RENCA cell line in BALB/c mice, generating an improved model of immunocompetent, metastatic renal cell carcinoma (RCC). Keywords: immunocompetent mouse model, ccRCC, RENCA, kidney cancer, CRISPR/Cas9, non-integrated lentivirus, immunotherapy

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

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