Strength of CAR signaling determines T cell versus ILC differentiation from pluripotent stem cells

Li, Suwen; Wang, Chloe S.; Montel‐Hagen, Amélie; Chen, Ho-Chung; Lopez, S. R.; Zhou, Olivia; Dai, Kristy; Tsai, Shau‐Wei; Satyadi, William; Botero, Carlos A.; Wong, Claudia; Casero, David; Crooks, Gay M.; Seet, Christopher S.

doi:10.1016/j.celrep.2023.112241

Cited by 8 publications

(8 citation statements)

References 84 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The iPSC-Tregs expressed typical markers of their primary natural Treg counterpart, namely highly expressing FOXP3 , IKZF2 and IL2RA in the absence of activation. The iPSC-Tregs also expressed a previously published 12 core Treg signature 39–41 similar to that of primary Tregs and less similar to iTregs and Tconv. Differential expression of genes between iPSC-Treg and primary Treg, such as IL7R and ID2 , could be due to culture conditions or to differences between cells that differentiated in vitro as compared to ex vivo adult cells.…”

Section: Discussionmentioning

confidence: 75%

“…While antigen specificity is unknown, reprogramming from a T cell clone possessing a single TCR 9,21,52 or introducing a CAR or transgenic TCR at the iPSC or T cell stage 8,11,12,22,53,54 can help confer specificity. Based on these findings, we sought to utilize a similar strategy to develop CD4sp T cells and Tregs from iPSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Using T cell-derived iPSCs may aid in more efficiently generating iPSC-T cells as the early stages of T cell development that are characterized by failed TCR rearrangement and cell death are bypassed 51 . While antigen specificity is unknown, reprogramming from a T cell clone possessing a single TCR 9, 21, 52 or introducing a CAR or transgenic TCR at the iPSC or T cell stage 8, 11, 12, 22, 53, 54 can help confer specificity. Based on these findings, we sought to utilize a similar strategy to develop CD4sp T cells and Tregs from iPSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Constitutive expression of CAR with simultaneous expression of the endogenous TCR, however, offsets the effects of a stronger Notch signal. In this case, both elimination of TRAC expression as well as attenuation of the CAR signal was required to restore DP T cell generation 11 ,12 . These are important considerations for the generation of a scalable cell therapy product using engineered iPSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Numerous studies have shown the ability to generate large quantities of T cells from iPSCs, focusing primarily on the differentiation to cytotoxic T lymphocytes (CTLs) [8][9][10] . Improvements have recently been made in the differentiation processes to generate iPSC-derived CAR CD8 single positive (CD8sp) T cells that more closely resemble primary CD8 + T cells rather than innate  T cells, by knocking out TRAC and delaying and attenuating CAR expression 11,12 . The artificial thymic organoid (ATO) system has also proven to be successful in generating functional CD3 + CD8 + T cells that produce IFN-, TNF-α, and IL-2 and proliferate upon stimulation 10 .…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Fong,

Mendel,

Jascur

et al. 2023

Preprint

View full text Add to dashboard Cite

iPSCs can serve as a renewable source of a consistent edited cell product, overcoming limitations of primary cells. While feeder-free generation of clinical grade iPSC-derived CD8 T cells has been achieved, differentiation of iPSC-derived CD4sp and regulatory T cells requires mouse stromal cells in an artificial thymic organoid. Here we report a serum- and feeder-free differentiation process suitable for large-scale production. Using an optimized concentration of PMA/Ionomycin, we generated iPSC-CD4sp T cells at high efficiency and converted them to Tregs using TGFβ and ATRA. Using zinc finger nucleases, we demonstrated high non-viral, targeted integration of an HLA-A2 CAR in iPSCs. iPSC-Tregs +/- HLA-A2 CAR phenotypically, transcriptionally and functionally resemble primary Tregs and suppress T cell proliferation in vitro. Our work is the first to demonstrate an iPSC-based platform amenable to manufacturing CD4 T cells to complement iPSC-CD8 oncology products and functional iPSC-Tregs to deliver Treg cell therapies at scale.

show abstract

Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Fong,

Mendel,

Jascur

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Unveiling the functional roles of patient‐derived tumour organoids in assessing the tumour microenvironment and immunotherapy

Chen,

Xu,

Xuan

et al. 2024

Clinical & Translational Med

View full text Add to dashboard Cite

Recent studies have established the pivotal roles of patient‐derived tumour organoids (PDTOs), innovative three‐dimensional (3D) culture systems, in various biological and medical applications. PDTOs, as promising tools, have been established and extensively used for drug screening, prediction of immune response and assessment of immunotherapeutic effectiveness in various cancer types, including glioma, ovarian cancer and so on. The overarching goal is to facilitate the translation of new therapeutic modalities to guide personalised immunotherapy. Notably, there has been a recent surge of interest in the co‐culture of PDTOs with immune cells to investigate the dynamic interactions between tumour cells and immune microenvironment. A comprehensive and in‐depth investigation is necessary to enhance our understanding of PDTOs as promising testing platforms for cancer immunotherapy. This review mainly focuses on the latest updates on the applications and challenges of PDTO‐based methods in anti‐cancer immune responses. We strive to provide a comprehensive understanding of the potential and prospects of PDTO‐based technologies as next‐generation strategies for advancing immunotherapy approaches.

show abstract

Rediscovering the human thymus through cutting-edge technologies

Pala,

Notarangelo,

Bosticardo

2024

Journal of Experimental Medicine

View full text Add to dashboard Cite

Recent technological advances have transformed our understanding of the human thymus. Innovations such as high-resolution imaging, single-cell omics, and organoid cultures, including thymic epithelial cell (TEC) differentiation and culture, and improvements in biomaterials, have further elucidated the thymus architecture, cellular dynamics, and molecular mechanisms underlying T cell development, and have unraveled previously unrecognized levels of stromal cell heterogeneity. These advancements offer unprecedented insights into thymic biology and hold promise for the development of novel therapeutic strategies for immune-related disorders.

show abstract

Strength of CAR signaling determines T cell versus ILC differentiation from pluripotent stem cells

Cited by 8 publications

References 84 publications

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Unveiling the functional roles of patient‐derived tumour organoids in assessing the tumour microenvironment and immunotherapy

Rediscovering the human thymus through cutting-edge technologies

Contact Info

Product

Resources

About