Allogeneic CAR T-Cell Editing via TRAC Locus Cleavage

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Solution Overview

Problem

Current treatments for relapsed or refractory B cell precursor acute lymphoblastic leukemia (BCP-ALL) face challenges such as high risk of graft-versus-host disease (GvHD) and limited effectiveness of existing CAR T cell therapies due to variability and insertional mutagenesis risks.

Innovation Solution

The method involves introducing guide RNA targeting the TRAC locus into αβ T cells from a donor, along with an RNA-guided nuclease and a homologous donor template encoding a CD19-specific chimeric antigen receptor (CAR), followed by expansion and administration of modified CAR T cells to the patient, which are produced from allogeneic donors to reduce GvHD risk and enhance antileukemic activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If allogeneic HSCT is used to treat high-risk leukemias, then graft-versus-leukemia effect is provided, but risk for graft-versus-host disease increases

Engineering Contradiction:

Improveantileukemic efficacyVSAvoidgraft-versus-host disease risk

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes αβ T-cells from the donor graft before transplantation, eliminating the primary source of graft-versus-host disease while preserving other beneficial cells such as CD34+ hematopoietic stem cells, NK cells, and γδ T-cells. This selective removal resolves the contradiction by eliminating the harmful component while maintaining the therapeutic benefits.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent applies different quality characteristics to different cell populations within the graft. By selectively eliminating αβ T-cells while preserving other cell types, the graft achieves local quality differentiation where specific cell populations have different functional properties - some providing antileukemic activity while others minimize GvHD risk.

Inventive Principle:

Principle #3Local quality

2Reliability

If donor leukocyte infusion is used to enhance antileukemic efficacy, then relapse prevention is improved, but severe graft-versus-host disease risk increases

Engineering Contradiction:

Improverelapse preventionVSAvoidsevere GvHD risk

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent removes αβ T-cells from the donor graft, eliminating the cells that would cause severe GvHD upon infusion. This allows subsequent infusion of donor-derived cells to provide antileukemic efficacy without the harmful alloreactive effects of conventional DLI.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent creates an asymmetric cell population in the graft by selectively depleting αβ T-cells while preserving other immune cells. This asymmetric composition allows the infused cells to provide one-sided benefit (antileukemic activity) without the reciprocal harm (GvHD) that characterizes conventional DLI.

Inventive Principle:

Principle #4Asymmetry

3Object-affected harmful factors

If αβ T-cells and CD19+ B-cells are selectively eliminated from graft, then GvHD incidence is reduced, but disease relapse becomes the most important cause of treatment failure

Engineering Contradiction:

ImproveGvHD incidenceVSAvoidtreatment success

Core Design Contradiction:

Object-affected harmful factorsVSReliability

Solution Approach 1:

The patent converts the harmful αβ T-cells into a beneficial therapeutic product by isolating them from the graft, genetically modifying them to express CAR receptors, and reinfusing them as engineered CAR T-cell products. This transforms the cells that would cause GvHD into cells that provide targeted antileukemic activity.

Inventive Principle:

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

Solution Approach 2:

The patent fundamentally changes the functional parameters of donor T-cells through genetic engineering. By introducing CAR receptors and modifying TCR expression, the cells' specificity and function are altered from potentially harmful alloreactive T-cells to beneficial CAR T-cells with targeted antileukemic activity and reduced GvHD risk.

Inventive Principle:

Principle #35Parameter changes

4Reliability

If patient-derived autologous T cells are used for CAR T cell therapy, then immune rejection risk is reduced, but manufacturing variability and insertional mutagenesis risk increase

Engineering Contradiction:

Improveimmune compatibilityVSAvoidproduct consistency

Core Design Contradiction:

ReliabilityVSManufacturing precision

Solution Approach 1:

The patent uses donor-derived T-cells as a template or copy source that can be consistently manufactured and expanded. These donor T-cells serve as a reliable starting material that can be genetically engineered with consistent CAR expression, reducing manufacturing variability while maintaining immune compatibility through HLA-matched donor selection.

Inventive Principle:

Principle #26Copying

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 effectively reduces leukemia burden while minimizing the risk of GvHD and insertional mutagenesis, providing a more reliable and targeted immune surveillance against leukemia without the need for lymphodepletion, and allows for the creation of multiple doses from a single manufacturing run.

Implementation Method 1

introducing into a plurality of αβ T cells from the donor a guide RNA targeting the TRAC locus, an RNA-guided nuclease, and a homologous donor template encoding a CD19-specific chimeric antigen receptor (CAR), wherein the RNA-guided nuclease cleaves the endogenous T cell receptor antigen constant (TRAC) locus

Methodology Applied

Scientific EffectRNA-guided nuclease cleavage: