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 journalArticle

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

Fingerprint

Natural Killer Cells
Immunotherapy
Clustered Regularly Interspaced Short Palindromic Repeats
Neoplasms
Genes
Dependovirus
Recombinant DNA
Homologous Recombination
Human Genome
Blood Cells
Cytokines

Keywords

  • ADAM17
  • CRISPR
  • NK cells
  • PD1
  • immunotherapy

PubMed: MeSH publication types

  • Journal Article

Cite this

A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy. / Pomeroy, Emily J.; Hunzeker, John T.; Kluesner, Mitchell G.; Lahr, Walker S.; Smeester, Branden A.; Crosby, Margaret R.; Lonetree, Cara lin; Yamamoto, Kenta; Bendzick, Laura; Miller, Jeffrey S.; Geller, Melissa A.; Walcheck, Bruce; Felices, Martin; Webber, Beau R.; Starr, Timothy K.; Moriarity, Branden S.

In: Molecular Therapy, Vol. 28, No. 1, 08.01.2020.

Research output: Contribution to journalArticle

Pomeroy, Emily J. ; Hunzeker, John T. ; Kluesner, Mitchell G. ; Lahr, Walker S. ; Smeester, Branden A. ; Crosby, Margaret R. ; Lonetree, Cara lin ; Yamamoto, Kenta ; Bendzick, Laura ; Miller, Jeffrey S. ; Geller, Melissa A. ; Walcheck, Bruce ; Felices, Martin ; Webber, Beau R. ; Starr, Timothy K. ; Moriarity, Branden S. / A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy. In: Molecular Therapy. 2020 ; Vol. 28, No. 1.
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