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A Three-dimensional Thymic Culture System to Generate Murine Induced Pluripotent Stem Cell-derived Tumor Antigen-specific Thymic Emigrants
Published on: August 9, 2019
Antigen-specific TCR-T cells from Rag2 gene-deleted pluripotent stem cells impede solid tumour growth in a mouse
Bingyan Wu1,2, Qi Zhang3, Pingshan Hong4
1CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
Abstract:
The technology of adoptive transfer of T-cell receptor (TCR) engineered T cells is wildly investigated as it has the potential to treat solid cancers. However, the therapeutic application of TCR-T cells is hampered by the poor quality derived mainly from patients' peripheral blood, as well as heterogeneous TCRs caused by the mismatch between transgenic and endogenous TCRs. To improve the homogeneity, antigen-specificity and reduce possible autoreactivity, here we developed a technique to generate antigen-specific T cells from Rag2 gene-deleted pluripotent stem cells (PSCs) and further measured their anti-tumour efficacy. PSCs were first targeted with OT1 TCR into the Rag2 locus to prevent TCR rearrangement during T-cell development. The engineered PSCs were then differentiated through a two-step strategy, in vitro generation of haematopoietic progenitor cells, and in vivo development and maturation of TCR-T cells. Finally, the response to tumour cells was assessed in vitro and in vivo. The regenerated OT1-iT displayed monoclonal antigen-specific TCR expression, and phonotypic normalities in the spleen and lymph node tissues. Importantly, the OT1-iT cells eliminated tumour cells while releasing specific cytokines in vitro. Furthermore, adoptive transfer of OT1-iT cells suppresses solid tumour growth in tumour-bearing animals. Our study presents a novel and straightforward strategy for producing antigen-specific TCR-T cells in vivo from PSCs, allowing for allogeneic transplantation and therapy of solid tumours.
Insights
This study introduces a new method to create uniform, cancer-targeting T-cell receptor (TCR) engineered T cells from stem cells. These engineered T cells effectively eliminate solid tumors, offering a promising new cancer therapy.
Area of Science:
- Immunology
- Cancer Biology
- Stem Cell Biology
Background:
- Adoptive transfer of T-cell receptor (TCR) engineered T cells shows promise for solid cancer treatment.
- Current TCR-T cell therapies face challenges due to poor cell quality and TCR heterogeneity from patient peripheral blood.
- Mismatched transgenic and endogenous TCRs can lead to reduced antigen specificity and potential autoreactivity.
Purpose of the Study:
- To develop a novel technique for generating homogeneous, antigen-specific T cells from gene-edited pluripotent stem cells (PSCs).
- To assess the anti-tumor efficacy of these engineered T cells in vitro and in vivo.
- To establish a potential strategy for allogeneic transplantation in solid tumor therapy.
Main Methods:
- Engineered PSCs by targeting the OT1 TCR into the Rag2 locus to ensure TCR homogeneity.
- Differentiated engineered PSCs through a two-step process: in vitro hematopoietic progenitor generation and in vivo T cell development.
- Assessed anti-tumor responses in vitro and in vivo, including cytokine release and tumor growth suppression.
Main Results:
- Generated antigen-specific T cells (OT1-iT) with monoclonal TCR expression and normal phenotypes.
- OT1-iT cells demonstrated effective tumor cell elimination and specific cytokine release in vitro.
- Adoptive transfer of OT1-iT cells significantly suppressed solid tumor growth in vivo.
Conclusions:
- Presents a novel and efficient strategy for generating antigen-specific TCR-T cells from PSCs.
- This method overcomes limitations of current TCR-T cell therapies, improving homogeneity and specificity.
- The approach holds potential for allogeneic transplantation and advanced solid tumor treatment.
