Development of off-the-shelf hematopoietic stem cell-engineered invariant natural killer T cells for COVID-19 therapeutic intervention

Yan-Ruide Li; Zachary Spencer Dunn; Gustavo Garcia Jr; Camille Carmona; Yang Zhou; Derek Lee; Jiaji Yu; Jie Huang; Jocelyn T Kim; Vaithilingaraja Arumugaswami; Pin Wang; Lili Yang

doi:10.1186/s13287-022-02787-2

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. doi: 10.1186/s13287-022-02787-2.

Authors

Yan-Ruide Li¹, Zachary Spencer Dunn², Gustavo Garcia Jr³, Camille Carmona¹, Yang Zhou¹, Derek Lee¹, Jiaji Yu¹, Jie Huang¹, Jocelyn T Kim⁴, Vaithilingaraja Arumugaswami^{3

5}, Pin Wang², Lili Yang^{6

7

8

9}

Affiliations

¹ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
² Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, Los Angeles, CA, 90089, USA.
³ Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁴ Division of Infectious Diseases, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁵ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁶ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁷ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁸ Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.

Abstract

Background: New COVID-19 treatments are desperately needed as case numbers continue to rise and emergent strains threaten vaccine efficacy. Cell therapy has revolutionized cancer treatment and holds much promise in combatting infectious disease, including COVID-19. Invariant natural killer T (iNKT) cells are a rare subset of T cells with potent antiviral and immunoregulatory functions and an excellent safety profile. Current iNKT cell strategies are hindered by the extremely low presence of iNKT cells, and we have developed a platform to overcome this critical limitation.

Methods: We produced allogeneic HSC-engineered iNKT (^AlloHSC-iNKT) cells through TCR engineering of human cord blood CD34⁺ hematopoietic stem cells (HSCs) and differentiation of these HSCs into iNKT cells in an Ex Vivo HSC-Derived iNKT Cell Culture. We then established in vitro SARS-CoV-2 infection assays to assess ^AlloHSC-iNKT cell antiviral and anti-hyperinflammation functions. Lastly, using in vitro and in vivo preclinical models, we evaluated ^AlloHSC-iNKT cell safety and immunogenicity for off-the-shelf application.

Results: We reliably generated ^AlloHSC-iNKT cells at high-yield and of high-purity; these resulting cells closely resembled endogenous human iNKT cells in phenotypes and functionalities. In cell culture, ^AlloHSC-iNKT cells directly killed SARS-CoV-2 infected cells and also selectively eliminated SARS-CoV-2 infection-stimulated inflammatory monocytes. In an in vitro mixed lymphocyte reaction (MLR) assay and an NSG mouse xenograft model, ^AlloHSC-iNKT cells were resistant to T cell-mediated alloreaction and did not cause GvHD.

Conclusions: Here, we report a method to robustly produce therapeutic levels of ^AlloHSC-iNKT cells. Preclinical studies showed that these ^AlloHSC-iNKT cells closely resembled endogenous human iNKT cells, could reduce SARS-CoV-2 virus infection load and mitigate virus infection-induced hyperinflammation, and meanwhile were free of GvHD-risk and resistant to T cell-mediated allorejection. These results support the development of ^AlloHSC-iNKT cells as a promising off-the-shelf cell product for treating COVID-19; such a cell product has the potential to target the new emerging SARS-CoV-2 variants as well as the future new emerging viruses.

Keywords: Allogeneic adoptive cell transfer; Coronavirus disease 2019 (COVID-19); Hematopoietic stem cell (HSC); Invariant natural killer T (iNKT) cell; Off-the-shelf cellular product; Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Publication types

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

MeSH terms

Animals
COVID-19* / therapy
Hematopoietic Stem Cells
Humans
Mice
Natural Killer T-Cells*
SARS-CoV-2

Supplementary concepts

SARS-CoV-2 variants

Abstract

Publication types

MeSH terms

Supplementary concepts

Grants and funding