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In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Naïve CD4+ T Cells Using a TGF-β-containing Protocol
Published on: December 30, 2016
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Gene editing to induce FOXP3 expression in human CD4+ T cells leads to a stable regulatory phenotype and function
Yuchi Honaker1, Nicholas Hubbard1, Yufei Xiang1
1Center for Immunity and Immunotherapies and the Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
Science Translational Medicine
|June 5, 2020
Summary
Researchers engineered T cells to express FOXP3, creating regulatory T cells (Tregs) that suppress autoimmune responses. These engineered Tregs show promise for cell therapy in treating inflammatory and autoimmune diseases.
Area of Science:
- Immunology
- Cell Biology
- Gene Editing
Background:
- Thymic regulatory T cells (tTregs) are crucial for preventing autoimmune diseases.
- Defects in tTreg number or function are linked to autoimmune conditions, driving interest in Treg-based cell therapies.
- Challenges exist in isolating and expanding sufficient functional tTregs for therapeutic use.
Purpose of the Study:
- To develop an alternative strategy for generating regulatory T cell-like cells (Tregs) from bulk CD4+ T cells.
- To engineer autologous T cells to stably express FOXP3, the key transcription factor for Treg function.
- To assess the therapeutic potential of these engineered Tregs (edTregs) in treating autoimmune and inflammatory diseases.
Main Methods:
- Utilized homology-directed repair (HDR) gene editing to insert an enhancer/promoter sequence near the FOXP3 gene.
- This strategy aimed to overcome epigenetic silencing and ensure robust endogenous FOXP3 expression.
- Evaluated the transcriptional profile, suppressive activity, and in vivo efficacy of engineered T cells (edTregs) in human and murine models.
Main Results:
- HDR-edited T cells (edTregs) demonstrated a stable transcriptional program characteristic of tTregs.
- edTregs exhibited potent suppressive activity in vitro and mediated immunosuppression in vivo in inflammatory disease models.
- The engineering approach successfully generated antigen-specific edTregs with significant functional capacity.
- Scalable enrichment and expansion methods for edTregs were established using clinically relevant techniques.
Conclusions:
- Engineered T cells (edTregs) effectively mimic the function of natural Tregs.
- This gene-editing strategy provides a viable method for producing functional, autologous Tregs for cell therapy.
- edTreg production holds significant potential for broad clinical application in managing autoimmune and inflammatory conditions.
