Maturation and persistence of CAR T cells derived from human pluripotent stem cells via chemical inhibition of G9a/GLP

Ran Jing; Marcelo Falchetti; Tianxiao Han; Mohamad Najia; Luca T Hensch; Eleanor Meader; Edroaldo Lummertz da Rocha; Martin Kononov; Stephanie Wang; Trevor Bingham; Zhiheng Li; Yunliang Zhao; Katie Frenis; Caroline Kubaczka; Song Yang; Deepak Jha; Gabriela F Rodrigues-Luiz; R Grant Rowe; Thorsten M Schlaeger; Marcela V Maus; Trista E North; Leonard I Zon; George Q Daley

doi:10.1016/j.stem.2024.10.004

Maturation and persistence of CAR T cells derived from human pluripotent stem cells via chemical inhibition of G9a/GLP

Cell Stem Cell. 2024 Nov 1:S1934-5909(24)00366-7. doi: 10.1016/j.stem.2024.10.004. Online ahead of print.

Authors

Ran Jing¹, Marcelo Falchetti¹, Tianxiao Han², Mohamad Najia¹, Luca T Hensch², Eleanor Meader², Edroaldo Lummertz da Rocha³, Martin Kononov², Stephanie Wang², Trevor Bingham², Zhiheng Li², Yunliang Zhao², Katie Frenis², Caroline Kubaczka², Song Yang², Deepak Jha², Gabriela F Rodrigues-Luiz⁴, R Grant Rowe⁵, Thorsten M Schlaeger², Marcela V Maus⁶, Trista E North⁷, Leonard I Zon⁸, George Q Daley⁹

Affiliations

¹ Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
² Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
³ Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, SC 88040-900, Brazil.
⁴ Graduate Program of Pharmacology, Center for Biological Sciences, Federal University of Santa Catarina, Florianópolis, SC 88040-900, Brazil.
⁵ Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Boston, MA 02115, USA.
⁶ Cellular Immunotherapy Program, Massachusetts General Hospital Cancer Center, Charlestown, MA 02114, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
⁷ Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA; Developmental and Regenerative Biology Program, Harvard Medical School, Boston, MA 02115, USA.
⁸ Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA; Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA.
⁹ Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: george.daley@childrens.harvard.edu.

PMID: 39504968
DOI: 10.1016/j.stem.2024.10.004

Abstract

Elucidating mechanisms of T cell development can guide in vitro T cell differentiation from induced pluripotent stem cells (iPSCs) and facilitate off-the-shelf T cell-based immunotherapies. Using a stroma-free human iPSC-T cell differentiation platform, we screened for epigenetic modulators that influence T cell specification and identified the H3K9-directed histone methyltransferases G9a/GLP as repressors of T cell fate. We show that G9a/GLP inhibition during specific time windows of differentiation of hematopoietic stem and progenitor cells (HSPCs) skews cell fates toward lymphoid lineages. Inhibition of G9a/GLP promotes the production of lymphoid cells during zebrafish embryonic hematopoiesis, demonstrating the evolutionary conservation of G9a/GLP function. Importantly, chemical inhibition of G9a/GLP facilitates the generation of mature iPSC-T cells that bear transcriptional similarity to peripheral blood αβ T cells. When engineered to express chimeric antigen receptors, the epigenetically engineered iPSC-T cells exhibit enhanced effector functions in vitro and durable, persistent antitumor activity in a xenograft tumor-rechallenge model.

Keywords: CAR-T cells; G9a/GLP; T cell differentiation; cancer immunotherapy; chemical screen; epigenetic regulation; hematopoiesis; lymphoid development; pluripotent stem cells.

Abstract

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