CAR-Engineered Inducible Regulatory T Cells for Stable Suppression
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Solution Overview
Problem
Existing regulatory T cells with FoxP3 defects or mutations face impaired differentiation and suppression functions, necessitating improved methods to enhance their stability and immunosuppressive capabilities.
Innovation Solution
Introduction of a chimeric antigen receptor (CAR) into inducible regulatory T cells, specifically through a multi-step process involving stimulation and dormant culture, to achieve high expression of suppression molecules and stable function.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If regulatory T cells are induced from peripheral T cells using conventional methods, then they can be produced, but their suppression function and stability are insufficient
Solution Approach 1:
The patent applies preliminary action by introducing the CAR gene into peripheral T cells before inducing regulatory T cell differentiation. This ensures that the suppression function is enhanced from the outset, rather than attempting to improve it after differentiation. The CAR gene integration occurs during the initial culture phase, allowing subsequent differentiation to proceed with pre-established genetic modifications that confer stable suppressive capabilities.
Solution Approach 2:
The patent employs parameter changes by modifying the culture conditions with specific cytokines (TGF-β, IL-2, IL-6, IL-10) and small molecule compounds (HDAC inhibitors, DNA methyltransferase inhibitors) to create an optimized induction environment. These parameter changes in the culture medium composition and signaling pathways enable the generation of regulatory T cells with enhanced and stable suppression function, overcoming the limitations of conventional induction methods.
2Stability of the object's composition
If FoxP3 expression is enhanced to improve suppression function, then regulatory T cell stability improves, but the complexity of the induction process increases
Solution Approach 1:
The patent uses small molecule compounds as intermediaries to bridge the gap between simple culture conditions and complex epigenetic modifications. Specifically, HDAC inhibitors and DNA methyltransferase inhibitors act as mediators that simplify the induction process by directly modifying chromatin structure and gene expression patterns, thereby enhancing FoxP3 stability without requiring complex multi-step differentiation protocols. These intermediary compounds facilitate epigenetic reprogramming in a controlled manner.
Solution Approach 2:
The patent applies composite materials by combining multiple cytokines (TGF-β, IL-2, IL-6, IL-10) and small molecule compounds in a synergistic culture medium formulation. This composite approach creates a unified induction system where each component contributes to different aspects of regulatory T cell differentiation and stabilization, achieving enhanced FoxP3 expression through the combined effect rather than relying on a single complex protocol.
3Reliability
If chimeric antigen receptor (CAR) is introduced into regulatory T cells, then suppression function is enhanced, but the difficulty of cell culture and maintenance increases
Solution Approach 1:
The patent merges the CAR gene introduction step with the regulatory T cell induction process into a single integrated workflow. Rather than separately generating regulatory T cells and then introducing CAR, the CAR gene is introduced into peripheral T cells at the beginning of the induction process, allowing simultaneous differentiation and genetic modification. This merging eliminates additional manipulation steps and simplifies the overall manufacturing process while maintaining enhanced immunosuppressive capability.
Solution Approach 2:
The patent ensures continuity of useful action by maintaining continuous culture conditions that support both CAR expression and regulatory T cell function throughout the induction and expansion phases. The culture medium is continuously supplemented with necessary cytokines and small molecule compounds, allowing the cells to progressively differentiate and maintain stable CAR expression without requiring repeated passaging or complex intervention, thereby easing manufacturing operations.
Data Source
Figure 1A~1B
Figure 2
Figure 3A~3B
AI summary
The present disclosure provides a pharmaceutical composition for treating or preventing a T cell-associated disease, including an inducible regulatory T cell having high functionality and a stable immunosuppressive function, and also provides inducible regulatory T cells containing a chimeric antigen receptor (CAR). In the present disclosure, there is provided a pharmaceutical composition for treating or preventing a T cell-associated disease, the pharmaceutical composition containing, as an active ingredient, inducible regulatory T cells having at least one characteristic selected from the group consisting of CTLA4 positive, NT5E positive, ITGAE (CD103) positive, and AREG positive. In the present disclosure, inducible regulatory T cells containing a chimeric antigen receptor (CAR) are provided.