Article Text

Download PDFPDF

417 Engineer off-the-shelf NKT cells for cancer immunotherapy
  1. Yanruide Li1 and
  2. Lili Yang2
  1. 1University of California, Los Angeles, North Hills, CA, USA
  2. 2University of California, Los Angeles, Los Angeles, CA, USA
  • Journal for ImmunoTherapy of Cancer (JITC) preprint. The copyright holder for this preprint are the authors/funders, who have granted JITC permission to display the preprint. All rights reserved. No reuse allowed without permission.


Background Cell-based immunotherapy has emerged as a promising approach in disease treatment, necessitating the development of readily available ’off-the-shelf’ cell products that can be manufactured at scale. Invariant natural killer T (iNKT) cells have shown potential as ideal cell carriers for allogeneic cell therapy, as they possess potent anti-cancer/virus properties without the risk of graft-versus-host disease (GvHD).1–4 However, the scarcity of endogenous iNKT cells in healthy donor blood poses a challenge.

Methods Here we present a novel strategy combining hematopoietic stem cell (HSC) gene engineering and ex vivo differentiation to produce high-yield, high-purity allogeneic HSC-engineered iNKT (AlloHSC-NKT) cells that closely resemble endogenous iNKT cells.

Results Our investigation focuses on three key aspects of AlloHSC-NKT cells: 1) Evaluating their antitumor capacity and exploring their multiple tumor targeting mechanisms; 2) Assessing their antiviral activity, particularly against the SARS-CoV-2 virus; 3) Investigating their immunoregulatory potential, with a specific emphasis on their ability to ameliorate GvHD. Furthermore, we explore the potential for enhancing the tumor-targeting capabilities of AlloHSC-NKT cells through chimeric antigen receptor (CAR) engineering, as well as reducing their immunogenicity by gene editing to ablate surface human leukocyte antigen (HLA) molecules.

Conclusions Collectively, these preclinical studies provide compelling evidence for the feasibility and therapeutic potential of AlloHSC-iNKT cell products and support their translation into clinical and commercial development.1–4


  1. Li YR, Zhou Y, Kim YJ, Zhu Y, Ma F, Yu J, Wang YC, Chen X, Li Z, Zeng S, Wang X, Lee D, Ku J, Tsao T, Hardoy C, Huang J, Cheng D, Montel-Hagen A, Seet CS, Crooks GM, Larson SM, Sasine JP, Wang X, Pellegrini M, Ribas A, Kohn DB, Witte O, Wang P, Yang L. Development of allogeneic HSC-engineered iNKT cells for off-the-shelf cancer immunotherapy. Cell Rep Med. 2021 Nov 16;2(11):100449.

  2. Li YR, Dunn ZS, Yu Y, Li M, Wang P, Yang L. Advancing cell-based cancer immunotherapy through stem cell engineering. Cell Stem Cell. 2023 May 4;30(5):592–610.

  3. Li YR, Dunn ZS, Garcia G Jr, Carmona C, Zhou Y, Lee D, Yu J, Huang J, Kim JT, Arumugaswami V, Wang P, Yang L. Development of off-the-shelf hematopoietic stem cell-engineered invariant natural killer T cells for COVID-19 therapeutic intervention. Stem Cell Res Ther. 2022 Mar 21;13(1):112.

  4. Li YR, Zeng S, Dunn ZS, Zhou Y, Li Z, Yu J, Wang YC, Ku J, Cook N, Kramer A, Yang L. Off-the-shelf third-party HSC-engineered iNKT cells for ameliorating GvHD while preserving GvL effect in the treatment of blood cancers. iScience. 2022 Aug 6;25(9):104859.

Ethics Approval Healthy donor human PBMCs were obtained from the UCLA/CFAR Virology Core Laboratory, without identification information under federal and state regulations. Patient bone marrow samples were collected following UCLA IRB approval (IRB#15–000062).

Consent Written informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.