A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy

Emily J. Pomeroy, John T. Hunzeker, Mitchell G. Kluesner, Walker S. Lahr, Branden A. Smeester, Margaret R. Crosby, Cara lin Lonetree, Kenta Yamamoto, Laura Bendzick, Jeffrey S. Miller, Melissa A. Geller, Bruce Walcheck, Martin Felices, Beau R. Webber, Timothy K. Starr, Branden S. Moriarity

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Enhancing natural killer (NK) cell cytotoxicity by blocking inhibitory signaling could lead to improved NK-based cancer immunotherapy. Thus, we have developed a highly efficient method for editing the genome of human NK cells using CRISPR/Cas9 to knock out inhibitory signaling molecules. Our method efficiently edits up to 90% of primary peripheral blood NK cells. As a proof-of-principle we demonstrate highly efficient knockout of ADAM17 and PDCD1, genes that have a functional impact on NK cells, and demonstrate that these gene-edited NK cells have significantly improved activity, cytokine production, and cancer cell cytotoxicity. Furthermore, we were able to expand cells to clinically relevant numbers, without loss of activity. We also demonstrate that our CRISPR/Cas9 method can be used for efficient knockin of genes by delivering homologous recombination template DNA using recombinant adeno-associated virus serotype 6 (rAAV6). Our platform represents a feasible method for generating engineered primary NK cells as a universal therapeutic for cancer immunotherapy.

Original languageEnglish (US)
JournalMolecular Therapy
Volume28
Issue number1
DOIs
StatePublished - Jan 8 2020

Bibliographical note

Funding Information:
T.S. is supported by funds from the Jan Chorzempa Cancer Research Endowed Fund, the Masonic Cancer Center's Translational Working Group Grant, the Randy Shaver Cancer Research and Community Fund, departmental funds, and institutional grants to MCC from NIH/NCI P30CA07759821 and CTSI from NCATS UL1TR00249402. We acknowledge the University of Minnesota Genomics Center, the Minnesota Supercomputing Institute, and the Masonic Cancer Center for their support of this research. B.S.M. is supported by funds from NIH/NCI R21CA216652 and U54CA232561 and the Children's Cancer Research Fund. T.K.S. is supported by funds from the Jan Chorzempa Cancer Research Endowed Fund, the Masonic Cancer Center's Translational Working Group Grant, the Randy Shaver Cancer Research and Community Fund, departmental funds, and institutional grants to MCC from NIH/NCI P30CA07759821 and CTSI from NCATS UL1TR00249402.

Funding Information:
T.S. is supported by funds from the Jan Chorzempa Cancer Research Endowed Fund, the Masonic Cancer Center's Translational Working Group Grant, the Randy Shaver Cancer Research and Community Fund , departmental funds, and institutional grants to MCC from  NIH/NCI P30CA07759821 and CTSI from NCATS UL1TR00249402 . We acknowledge the University of Minnesota Genomics Center , the Minnesota Supercomputing Institute , and the Masonic Cancer Center for their support of this research. B.S.M. is supported by funds from NIH/NCI R21CA216652 and U54CA232561 and the Children's Cancer Research Fund. T.K.S. is supported by funds from the Jan Chorzempa Cancer Research Endowed Fund, the Masonic Cancer Center’s Translational Working Group Grant, the Randy Shaver Cancer Research and Community Fund, departmental funds, and institutional grants to MCC from NIH/NCI P30CA07759821 and CTSI from NCATS UL1TR00249402 .

Publisher Copyright:
© 2019 The American Society of Gene and Cell Therapy

Keywords

  • ADAM17
  • CRISPR
  • NK cells
  • PD1
  • immunotherapy

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