CRISPR Guide RNAs for T-Cell Inhibitory Receptor Disruption

Solution Overview

Problem

Current cancer immunotherapy methods, such as adoptive T cell therapy (ACT) using genetically engineered T cells, face limitations due to short T cell persistence and T cell exhaustion, which is exacerbated by inhibitory receptors (IRs) upregulated in the tumor microenvironment, leading to reduced efficacy and potential autoimmune side effects.

Innovation Solution

Genetic disruption of inhibitory receptors (IRs) in T cells using CRISPR/Cas9 technology, combined with lentiviral vectors, to redirect T cell specificity against tumor antigens and permanently disrupt IRs, enhancing long-term persistence and anti-tumor activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If adoptive T cell therapy (ACT) with genetically engineered T cells is used, then tumor-specific T cell recognition and killing ability is improved, but T cell persistence and resistance to exhaustion are reduced

Engineering Contradiction:

ImproveT cell recognition and killing abilityVSAvoidT cell persistence

Core Design Contradiction:

ReliabilityVSDuration of action of moving object

Solution Approach 1:

The patent extracts and removes inhibitory receptors (IRs) from T cells using CRISPR/Cas9 gene editing technology. By taking out these harmful elements (IRs that cause exhaustion), the T cells gain enhanced persistence and resistance to exhaustion while maintaining their tumor-specific recognition and killing abilities.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the genetic parameters of T cells by disrupting the expression of inhibitory receptors through CRISPR/Cas9-mediated gene editing. This parameter change (removing IR expression) fundamentally alters the T cell's interaction with the tumor microenvironment, enabling long-term persistence and sustained anti-tumor activity.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If adoptive T cell therapy (ACT) with genetically engineered T cells is used, then tumor-specific T cell recognition and killing ability is improved, but T cell exhaustion is exacerbated

Engineering Contradiction:

ImproveT cell recognition and killing abilityVSAvoidT cell exhaustion

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent extracts and removes inhibitory receptors (IRs) from T cells using CRISPR/Cas9 gene editing technology. By taking out these harmful elements (IRs that cause exhaustion), the T cells gain enhanced persistence and resistance to exhaustion while maintaining their tumor-specific recognition and killing abilities.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the harmful effect of inhibitory receptors (which normally cause T cell exhaustion) into a benefit by using CRISPR/Cas9 to precisely remove them. This transformation turns the previously detrimental IR expression into complete absence, thereby converting the exhaustion problem into enhanced T cell resilience and persistence.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Object-generated harmful factors

If inhibitory receptors (IRs) are upregulated in the tumor microenvironment, then T cell exhaustion is promoted, but tumor escape is enabled

Engineering Contradiction:

ImproveT cell exhaustionVSAvoidAnti-tumor immunity

Core Design Contradiction:

Object-generated harmful factorsVSReliability

Solution Approach 1:

The patent extracts and removes inhibitory receptors (IRs) from T cells using CRISPR/Cas9 gene editing technology. By taking out these harmful elements (IRs that cause exhaustion), the T cells gain enhanced persistence and resistance to exhaustion while maintaining their tumor-specific recognition and killing abilities.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the harmful effect of inhibitory receptors (which normally cause T cell exhaustion) into a benefit by using CRISPR/Cas9 to precisely remove them. This transformation turns the previously detrimental IR expression into complete absence, thereby converting the exhaustion problem into enhanced T cell resilience and persistence.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

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

CRISPR/Cas9-mediated IR disruptions in T cells achieve high efficiency in targeting tumor antigens, maintaining effector functions, and resisting exhaustion, demonstrating enhanced cytotoxicity and long-lasting anti-tumor responses in preclinical models.

Implementation Method 1

Genetic disruption of inhibitory receptors (IRs) in T cells using CRISPR/Cas9 technology

Methodology Applied

Scientific EffectCRISPR/Cas9 gene editing:

Implementation Method 2

CRISPR/Cas9 system comprising guide RNAs directed to at least one inhibitory receptor (IR) gene

Methodology Applied

Scientific EffectGuide RNA-directed nuclease targeting:

Implementation Method 3

combined with lentiviral vectors, to redirect T cell specificity against tumor antigens

Methodology Applied

Scientific EffectViral transduction:

Implementation Method 4

redirect T cell specificity against tumor antigens

Methodology Applied

Scientific EffectT cell antigen recognition:

Implementation Method 5

demonstrating enhanced cytotoxicity and long-lasting anti-tumor responses

Methodology Applied

Scientific EffectCytotoxic T cell killing:

Data Source

PatentUS20250270546A1Guide rnas and uses thereof

Publication Date: 2025.08.28 FOND CENT SAN RAFFAELE

AI summary

The present invention is related to an isolated guide ribonucleic acid (gRNA) including a guide sequence targeting an inhibitory receptor (IR), a TCR α (TRAC) constant region or a β chain (TRBC1/2) constant region target sequence, wherein the guide sequence is selected from the group consisting of SEQ ID NOs: 1-27, 122-126 and combinations thereof.