Hypoimmunogenic Stem Cells via CRISPR-Engineered Immune Inhibitors

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Solution Overview

Problem

Current stem cell therapies face challenges with immune rejection when transplanting cells, tissues, and organs, particularly in allogeneic settings, due to recognition by immune cells like NK cells and T cells, necessitating alternative strategies to inhibit immune activation.

Innovation Solution

Artificially engineered stem cells with increased expression or activity of immune activity inhibitory factors such as LGALS9, CLEC4G, PD-L1, and HHLA2 are introduced to inhibit immune rejection by modifying the genome of stem cells using gene editing technologies like CRISPR/Cas9, ensuring hypoimmunogenicity and immune tolerance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If allogeneic stem cells are used for cell therapy, then commercialization is enabled and production time is reduced, but immune rejection occurs due to recognition by NK cells and T cells

Engineering Contradiction:

Improveproduction timeVSAvoidimmune rejection

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent applies preliminary anti-action by pre-engineering stem cells to express immune inhibitory factors (PD-L1, HHLA2, LGALS9, CLEC4G) before transplantation. These factors are introduced into the stem cell genome using CRISPR/Cas9 technology, enabling the cells to proactively inhibit immune rejection mechanisms (PD-1/PD-L1, KIR/HHLA2, TIM-3/Gal-9 pathways) before encountering the immune system, thus preventing immune activation while maintaining allogeneic production advantages

Inventive Principle:

Principle #9Preliminary anti-action

Solution Approach 2:

The patent changes the biological parameters of stem cells by modifying their gene expression profiles. Specifically, it increases the expression levels of immune inhibitory factors (PD-L1, HHLA2, LGALS9, CLEC4G) through genetic engineering, transforming the cells from immunogenic to hypoimmunogenic states. This parameter change enables the cells to evade immune recognition and rejection while maintaining their therapeutic potential

Inventive Principle:

Principle #35Parameter changes

2Object-affected harmful factors

If autologous stem cells are used for cell therapy, then immune rejection is minimized, but production time and manpower requirements increase

Engineering Contradiction:

Improveimmune rejectionVSAvoidproduction time

Core Design Contradiction:

Object-affected harmful factorsVSLoss of time

Solution Approach 1:

The patent uses copying by creating allogeneic stem cell lines that replicate the successful autologous approach at a scaled-up level. Instead of customizing each patient's cells individually, the patent establishes standardized allogeneic cell lines with engineered immune inhibitory factors that can be produced in bulk and distributed to multiple patients, effectively copying the immune-protective mechanism while eliminating the time-consuming personalized production process

Inventive Principle:

Principle #26Copying

3Object-affected harmful factors

If immune inhibitory factors are engineered into stem cells, then immune rejection is inhibited, but genetic engineering complexity increases

Engineering Contradiction:

Improveimmune rejectionVSAvoidgenetic engineering complexity

Core Design Contradiction:

Object-affected harmful factorsVSDevice complexity

Solution Approach 1:

The patent applies segmentation by dividing the immune inhibition function into multiple independent genetic modules. Instead of creating a single complex inhibitor, the patent introduces separate genes for different immune inhibitory factors (PD-L1 for PD-1 pathway, HHLA2 for KIR pathway, LGALS9 for TIM-3 pathway, CLEC4G for LAG-3 pathway), allowing each factor to independently suppress specific immune activation routes. This modular approach simplifies the genetic engineering process and enables comprehensive immune inhibition through combination of simpler elements

Inventive Principle:

Principle #1Segmentation

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The modified stem cells exhibit reduced immune rejection, enabling their use as a therapeutic agent for cell transplantation and tissue regeneration without causing immune responses in autologous or allogeneic environments, achieving immune tolerance and enhanced expression of immune inhibitory factors.

Implementation Method 1

modifying the genome of stem cells using gene editing technologies like CRISPR/Cas9

Methodology Applied

Scientific EffectCRISPR/Cas9 gene editing:

Implementation Method 2

immune checkpoint receptors such as CTLA-4 and PD-1 on T cells send inhibitory signals that inactivate T cells through binding to immune checkpoint ligands such as CD80, CD86, and PD-L1 on abnormal cells

Methodology Applied

Scientific EffectImmune checkpoint inhibition:

Implementation Method 3

HHLA2 is a newly identified B7 family member that regulates T cell function. HHLA2 binds to killer cell immunoglobulin-like receptor, three immunoglobulin domains and long cytoplasmic tail 3 (KIR3DL3), a receptor on T and NK cells, and also inhibits T cell activation

Methodology Applied

Scientific EffectKIR-HHLA2 interaction:

Data Source

PatentEP4488364A1Low immunogenic stem cells, low immunogenic cells differentiated or derived from stem cells, and production method therefor

Publication Date: 2025.01.08 TOOLGEN INC

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AI summary

Provided are hypoimmunogenic stem cells, hypoimmunogenic cells differentiated or derived from stem cells capable of inhibiting immune rejection upon transplantation by increasing the expression or activity level of immune activity inhibitory factors, a method of preparing the same, and a cell therapeutic agent using the same.