Logic-Gated CAR-T Cells With Co-Expressed shRNA for Dual-Antigen Targeting

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Solution Overview

Problem

Current CAR-T cell-based immunotherapy for cancer faces challenges such as off-target toxicity, reduced expansion and effector function, susceptibility to suppression and exhaustion, and lack of memory T cell persistence.

Innovation Solution

Development of recombinant nucleic acids encoding chimeric priming receptors and chimeric antigen receptors, along with shRNA sequences targeting FAS, PTPN2, and/or TOX, to enhance the specificity and functionality of immune cells, including the use of non-viral vectors for genetic modification.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If CAR-T cells are engineered to target cancer cells, then cancer targeting ability is improved, but off-target toxicity increases

Engineering Contradiction:

Improvecancer targeting abilityVSAvoidoff-target toxicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent divides the targeting function into two separate components: a priming receptor that binds to a first antigen and a CAR that binds to a second antigen. Both receptors must be activated simultaneously for full T cell activation, creating a logical AND gate that ensures specificity to cancer cells expressing both antigens while avoiding off-target toxicity to cells expressing only one antigen

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent implements dynamic control of CAR expression through a logic gate system where CAR expression is induced only upon simultaneous binding of both priming receptor and CAR to their respective antigens. This dynamic regulation allows the T cells to remain dormant in normal tissues and activate only when encountering cancer cells with the specific dual-antigen signature

Inventive Principle:

Principle #15Dynamics

2Productivity

If CAR-T cells are activated to attack tumor cells, then tumor cell killing is improved, but T cell exhaustion and suppression increase

Engineering Contradiction:

Improvetumor cell killingVSAvoidT cell persistence

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent performs preliminary genetic modification of T cells to include shRNA sequences targeting exhaustion-related genes (FAS, PTPN2, TOX) before T cell activation and therapy administration. This preliminary action prevents the development of exhaustion and suppression during the anti-tumor response, maintaining T cell functionality and persistence throughout treatment

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent implements feedback control by monitoring T cell activation signals and regulating the expression of exhaustion-related genes accordingly. The logic gate system provides feedback mechanisms that control CAR expression and T cell activation states, preventing premature exhaustion while maintaining sustained anti-tumor activity

Inventive Principle:

Principle #23Feedback

3Reliability

If shRNA sequences are introduced to reduce exhaustion, then T cell persistence is improved, but genetic modification complexity increases

Engineering Contradiction:

ImproveT cell persistenceVSAvoidgenetic modification complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent merges multiple functions into a single integrated genetic construct that simultaneously provides: (1) priming receptor expression, (2) CAR expression under logic gate control, and (3) shRNA-mediated knockdown of exhaustion genes. This consolidation reduces the number of separate genetic modifications needed while achieving multiple therapeutic goals

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent creates universal T cell engineering platforms where a single genetic modification strategy simultaneously addresses targeting specificity (through logic gate-controlled CAR expression) and T cell persistence (through shRNA-mediated exhaustion gene knockdown), making the approach broadly applicable to different cancer types and antigen targets

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

The proposed solution significantly reduces off-target toxicity and enhances the survival, expansion, and effector function of CAR-T cells, leading to improved cancer targeting and memory T cell persistence.

Implementation Method 1

at least one nucleic acid sequence at least 15 nucleotides in length, wherein the at least one nucleic acid sequence comprises one or more of: (1) a first nucleic acid sequence complementary to nucleotides 1126 to 1364 of an mRNA encoding human Fas Cell Surface Death Receptor (FAS)... (2) a second nucleic acid sequence complementary to nucleotides 518 to 559 of an mRNA encoding human Protein Tyrosine Phosphatase Non-Receptor Type 2 (PTPN2)... and (3) a third nucleic acid sequence complementary to nucleotides 1294 to 2141 of an mRNA encoding human Thymocyte Selection Associated High Mobility Group Box (TOX)

Methodology Applied

Scientific EffectRNA interference:

Data Source

PatentUS20250250350A1Immune cells having co-expressed shrnas and logic gate systems

Publication Date: 2025.08.07 ARSENAL BIOSCIENCES INC

AI summary

Provided herein are recombinant nucleic acids encoding chimeric priming receptors that bind ALPG/P, chimeric antigen receptors that bind MSLN, and shRNA that target FAS, PTPN2, and/or TOX. Also provided are systems of chimeric priming receptors that bind ALPG/P, chimeric antigen receptors that bind MSLN, and shRNA that target FAS, PTPN2, and/or TOX, cells expressing such proteins and shRNA, and methods of use thereof.