Development of allogeneic HSC-engineered iNKT cells for off-the-shelf cancer immunotherapy

Cell Rep Med. 2021 Nov 16;2(11):100449. doi: 10.1016/j.xcrm.2021.100449.


Cell-based immunotherapy has become the new-generation cancer medicine, and "off-the-shelf" cell products that can be manufactured at large scale and distributed readily to treat patients are necessary. Invariant natural killer T (iNKT) cells are ideal cell carriers for developing allogeneic cell therapy because they are powerful immune cells targeting cancers without graft-versus-host disease (GvHD) risk. However, healthy donor blood contains extremely low numbers of endogenous iNKT cells. Here, by combining hematopoietic stem cell (HSC) gene engineering and in vitro differentiation, we generate human allogeneic HSC-engineered iNKT (AlloHSC-iNKT) cells at high yield and purity; these cells closely resemble endogenous iNKT cells, effectively target tumor cells using multiple mechanisms, and exhibit high safety and low immunogenicity. These cells can be further engineered with chimeric antigen receptor (CAR) to enhance tumor targeting or/and gene edited to ablate surface human leukocyte antigen (HLA) molecules and further reduce immunogenicity. Collectively, these preclinical studies demonstrate the feasibility and cancer therapy potential of AlloHSC-iNKT cell products and lay a foundation for their translational and clinical development.

Keywords: CAR-engineered conventional αβ T cells; HLA-ablated universal HSC-iNKT cells; allogeneic HSC-engineered iNKT cells; allogeneic off-the-shelf cell therapy; allorejection; cancer immunotherapy; chimeric antigen receptor; graft-versus-host disease; hematopoietic stem cell; invariant natural killer T cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allogeneic Cells / immunology*
  • Allogeneic Cells / metabolism
  • Animals
  • Cell Engineering*
  • Cell Line, Tumor
  • Gene Expression Profiling
  • HLA Antigens / metabolism
  • Hematopoietic Stem Cells / immunology*
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Immunotherapy*
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Natural Killer T-Cells / immunology*
  • Natural Killer T-Cells / metabolism
  • Neoplasms / immunology*
  • Neoplasms / therapy*
  • Phenotype
  • Receptors, Chimeric Antigen / metabolism
  • Transcriptome / genetics


  • HLA Antigens
  • Receptors, Chimeric Antigen