Regulatory T Cell Lineage Stability via Subset Segmentation

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Solution Overview

Problem

Current challenges in adoptive T cell therapy with regulatory T cells include the instability of T REG cells under inflammatory conditions, impurities of non-T REG effector T cells, and the difficulty in manufacturing pure T REG cell products.

Innovation Solution

A composition comprising CD4+CD25+CD127low regulatory T cells, enriched with naive, naive-like memory, and central memory regulatory T cell subsets, while excluding effector memory regulatory T cells, to enhance lineage stability and immunoregulatory potency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If bulk T REG cell populations are used for adoptive immunotherapy, then the manufacturing process is simpler, but the lineage stability under inflammatory conditions deteriorates

Engineering Contradiction:
Improvemanufacturing simplicityVSAvoidlineage stability
Core Design Contradiction:
Ease of manufactureVSStability of the object's composition

Solution Approach 1:

The patent segments the bulk T REG cell population into distinct subsets based on memory phenotype markers (CD45RA, CCR7, CD62L). It identifies and isolates specific subsets (naive T REG cells: CD45RA+CCR7+CD62L+; central memory T REG cells: CD45RO+CCR7+CD62L+) that demonstrate enhanced lineage stability. This segmentation allows selection of stable subsets while simplifying manufacturing by focusing on defined phenotypic criteria rather than complex functional assays.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent applies local quality by assigning different functional properties to different T REG cell subsets. Specifically, it identifies that naive and central memory T REG cell subsets possess superior lineage stability and immunoregulatory potency compared to effector memory subsets. By enriching for these specific subsets with defined phenotypic markers, the patent creates a product with localized enhanced quality in terms of stability and efficacy.

Inventive Principle:
Principle #3Local quality

2Reliability

If T REG cell products are purified to high purity, then safety and efficacy are improved, but the manufacturing complexity increases

Engineering Contradiction:
Improvesafety and efficacyVSAvoidmanufacturing complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent performs preliminary action by pre-enriching T REG cells from peripheral blood mononuclear cells using magnetic beads conjugated to anti-CD25 antibodies before further subset selection. This preliminary purification step reduces the burden on subsequent subset enrichment steps and removes most non-T REG cells early in the manufacturing process, thereby improving safety while managing overall manufacturing complexity.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent replaces complex mechanical separation methods with antibody-based magnetic bead enrichment and flow cytometry-based sorting. These biochemical and optical methods provide higher purity and more precise subset isolation with simpler operational procedures compared to traditional mechanical cell separation techniques.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Quantity of substance

If effector memory T REG cells are included in the product, then the cell yield is higher, but the immunoregulatory potency and stability deteriorate

Engineering Contradiction:
Improvecell yieldVSAvoidimmunoregulatory potency
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent changes the phenotypic parameters used to define the T REG cell product by specifying memory status markers (CD45RA, CCR7, CD62L expression patterns). By defining the product in terms of naive and central memory phenotypes rather than total T REG cell count, the patent ensures enhanced immunoregulatory potency and lineage stability while accepting that cell yield may be reduced compared to including all T REG subsets.

Inventive Principle:
Principle #35Parameter changes

Data Source

PatentEP4538366A1Enhancing lineage stability of therapeutic regulatory t cell products
Publication Date: 2025.04.16 CHARITE UNIVS MEDIZIN BERLIN
  • EP4538366A1 patent drawingFigure 1A~1B
  • EP4538366A1 patent drawingFigure 1C~1E
  • EP4538366A1 patent drawingFigure 2A~2G

AI summary

The present invention relates to the field of immunotherapy, in particular, to adoptive T cell therapy with regulatory T cells. The invention provides a composition comprising CD4+CD25+CD127low regulatory T cells having an enhanced lineage stability to inflammatory triggers, as well as methods of preparing such compositions, as well as methods producing regulatory T cell products by expanding such compositions. Regulatory T cell products obtained according to the method of the invention can be used e.g., for treating a subject having or at risk of having an undesired immune response, such as a subject having an autoimmune disease, an allergy or other overwhelming inflammatory diseases, a transplant or a gene therapy recipient.