Adoptive Transfer of NKG2D CAR mRNA-Engineered Natural Killer Cells in Colorectal Cancer Patients

Mol Ther. 2019 Jun 5;27(6):1114-1125. doi: 10.1016/j.ymthe.2019.03.011. Epub 2019 Mar 20.


By fusing the extracellular domain of the natural killer (NK) cell receptor NKG2D to DAP12, we constructed a chimeric antigen receptor (CAR) to improve NK cell tumor responses. An RNA electroporation approach that provides transient expression of the CAR was adopted as a risk mitigation strategy. Expression of the NKG2D RNA CAR significantly augmented the cytolytic activity of NK cells against several solid tumor cell lines in vitro and provided a clear therapeutic benefit to mice with established solid tumors. Three patients with metastatic colorectal cancer were then treated with local infusion of the CAR-NK cells. Reduction of ascites generation and a marked decrease in number of tumor cells in ascites samples were observed in the first two patients treated with intraperitoneal infusion of low doses of the CAR-NK cells. The third patient with metastatic tumor sites in the liver was treated with ultrasound-guided percutaneous injection, followed by intraperitoneal infusion of the CAR-NK cells. Rapid tumor regression in the liver region was observed with Doppler ultrasound imaging and complete metabolic response in the treated liver lesions was confirmed by positron emission tomography (PET)- computed tomographic (CT) scanning. Our results highlight a promising therapeutic potential of using RNA CAR-modified NK cells to treat metastatic colorectal cancer.

Keywords: NKG2D ligands; chimeric antigen receptor; colorectal cancer; immunotherapy; natural killer cell.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer / adverse effects
  • Adoptive Transfer / methods*
  • Animals
  • Cell Engineering / methods
  • Cell Transplantation / adverse effects
  • Cell Transplantation / methods*
  • Colorectal Neoplasms / mortality
  • Colorectal Neoplasms / pathology
  • Colorectal Neoplasms / therapy*
  • Cytotoxicity, Immunologic / genetics
  • Feasibility Studies
  • Female
  • Genetic Vectors
  • HCT116 Cells
  • Humans
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / metabolism
  • Killer Cells, Natural / transplantation*
  • Lymphocyte Activation
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Middle Aged
  • NK Cell Lectin-Like Receptor Subfamily K / genetics*
  • Pilot Projects
  • RNA, Messenger / genetics
  • Receptors, Chimeric Antigen / genetics
  • Receptors, Chimeric Antigen / immunology*
  • Receptors, Chimeric Antigen / metabolism
  • Treatment Outcome
  • Xenograft Model Antitumor Assays


  • KLRK1 protein, human
  • NK Cell Lectin-Like Receptor Subfamily K
  • RNA, Messenger
  • Receptors, Chimeric Antigen