Allogeneic T Cell Engineering via CRISPR Cas9

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Solution Overview

Problem

Current methods for adoptive immunotherapy using autologous T cells are hindered by technical and logistical challenges, including high costs, the need for dedicated facilities, and variability in efficacy and safety, while allogeneic cell therapies face rejection and graft-versus-host disease issues.

Innovation Solution

The use of RNA-guided endonucleases, such as the Cas9/CRISPR system, to genetically modify allogeneic T cells by inactivating specific genes and introducing chimeric antigen receptors (CARs) to redirect immune activity, enabling the production of non-alloreactive T cells suitable for immunotherapy.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If autologous T cells are used for adoptive immunotherapy, then the therapy can be personalized to the patient, but the process is expensive, requires dedicated facilities, and has variable efficacy and safety

Engineering Contradiction:

Improveefficacy and safety consistencyVSAvoiddedicated facilities and process complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent uses allogeneic T cells as copies of autologous therapy concepts, creating a standardized product that replicates the benefits of personalized therapy without requiring patient-specific processing facilities. The T cells are generated from donor sources and standardized through controlled genetic modification protocols.

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent modifies T cell parameters through CRISPR/Cas9-mediated genetic engineering, specifically inactivating endogenous TCR genes and introducing CARs with standardized parameters. This transforms variable autologous therapy into a standardized product with consistent efficacy and safety profiles.

Inventive Principle:

Principle #35Parameter changes

2Ease of manufacture

If allogeneic T cells are used for immunotherapy, then the therapy can be standardized and produced off-the-shelf, but the cells are rejected by the host or cause graft-versus-host disease

Engineering Contradiction:

Improvestandardized productionVSAvoidhost rejection and graft-versus-host disease

Core Design Contradiction:

Ease of manufactureVSObject-affected harmful factors

Solution Approach 1:

The patent extracts the harmful alloreactive TCR function from the allogeneic T cells by inactivating endogenous TCR genes through CRISPR/Cas9. This removes the source of host rejection and graft-versus-host disease while preserving the therapeutic potential of the allogeneic cells.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

Instead of trying to make allogeneic cells compatible with the host through traditional methods, the patent inverts the approach by actively removing the harmful TCR function and replacing it with CARs that target tumor antigens, transforming the cells into non-alloreactive therapeutic units.

Inventive Principle:

Principle #13The other way round (Inversion)

3Object-affected harmful factors

If T cell receptors are inactivated to prevent alloreactivity, then graft-versus-host disease is prevented, but T cell function is lost

Engineering Contradiction:

Improvegraft-versus-host disease preventionVSAvoidT cell function

Core Design Contradiction:

Object-affected harmful factorsVSReliability

Solution Approach 1:

The patent introduces CARs as intermediary molecules that replace the traditional TCR function. These CARs serve as mediators that enable T cells to recognize and respond to tumor antigens without requiring functional endogenous TCRs, thus preventing GVHD while maintaining therapeutic function.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent creates multi-functional T cells that can perform multiple functions: preventing alloreactivity through TCR inactivation, avoiding GVHD, and maintaining anti-tumor activity through CAR expression. This universal approach resolves the conflict between preventing harmful functions and maintaining therapeutic functions.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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

This approach allows for the precise modification of T cells, enabling their use as an 'off-the-shelf' therapeutic product for treating cancer, infections, and autoimmune diseases with reduced toxicity and increased efficacy, avoiding the limitations of autologous and allogeneic cell therapies.

Implementation Method 1

The use of RNA-guided endonucleases, such as the Cas9/CRISPR system, to genetically modify allogeneic T cells by inactivating specific genes

Methodology Applied

Scientific EffectCRISPR/Cas9 genome editing:

Implementation Method 2

The engineered T cells are also intended to express chimeric antigen receptors (CAR) to redirect their immune activity towards malignant or infected cells

Methodology Applied

Scientific EffectAntigen recognition and T-cell redirection:

Data Source

PatentUS11959091B2Methods for engineering T cells for immunotherapy by using RNA-guided Cas nuclease system

Publication Date: 2024.04.16 CELLECTIS SA

AI summary

The present invention relates to methods of developing genetically engineered, preferably non-alloreactive T-cells for immunotherapy. This method involves the use of RNA-guided endonucleases, in particular Cas9/CRISPR system, to specifically target a selection of key genes in T-cells. The engineered T-cells are also intended to express chimeric antigen receptors (CAR) to redirect their immune activity towards malignant or infected cells. The invention opens the way to standard and affordable adoptive immunotherapy strategies using T-Cells for treating cancer and viral infections.