PEBL-Mediated CD2 Blockade in CAR-T Cells

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Solution Overview

Problem

The variability in the fitness of immune cells collected from cancer patients complicates the identification of effective cell dosages for CAR-T cell therapy, leading to unpredictable therapeutic outcomes and risks of graft-versus-host disease in allogeneic settings.

Innovation Solution

The development of Protein Expression Blockers (PEBLs) that intracellularly bind CD2, preventing its surface expression and incorporating these into CAR-T cells to enhance their therapeutic efficacy and consistency, while reducing the risk of graft-versus-host disease.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If allogeneic T cells are used to improve consistency and effectiveness, then therapeutic reliability is improved, but graft-versus-host disease risk increases

Engineering Contradiction:

Improvetherapeutic consistencyVSAvoidgraft-versus-host disease

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes the harmful capability of allogeneic T cells to recognize and attack host tissues by deleting or modifying genes encoding T cell receptors and other immune recognition molecules. This extraction of harmful function allows the cells to maintain their proliferative and cytotoxic capabilities while eliminating the risk of graft-versus-host disease

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the inherent aggressiveness and immune recognition capabilities of allogeneic T cells, which normally cause graft-versus-host disease, into beneficial anti-tumor activity. By redirecting these cells with CARs and enhancing their proliferation, the same aggressive properties that caused harm are now harnessed to attack cancer cells

Inventive Principle:

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

2Reliability

If CAR-T cells are engineered to target specific antigens, then tumor cell killing is improved, but fratricide of CAR-T cells occurs when targeting CD2

Engineering Contradiction:

Improvetumor cell killingVSAvoidfratricide

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

Instead of trying to prevent CAR-T cells from recognizing CD2, the patent inverts the approach by making the CAR-T cells themselves CD2-negative through the PEBL mechanism. This inversion allows the CAR-T cells to selectively kill CD2-positive tumor cells while being protected from fratricide, as the target antigen is absent from the effector cells

Inventive Principle:

Principle #13The other way round (Inversion)

3Measurement precision

If variable fitness of immune cells is addressed to improve predictability, then dosing accuracy is improved, but cell selection becomes more complex

Engineering Contradiction:

Improvedosing accuracyVSAvoidcell processing complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent applies parameter changes by standardizing the functional state of T cells through controlled expansion protocols and genetic engineering. By adjusting parameters such as proliferation control elements, cytokine support, and genetic modifications, the patent creates a standardized cell product with predictable dosing requirements, transforming variable immune cells into a consistent therapeutic product

Inventive Principle:

Principle #35Parameter changes

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

PEBLs enable durable blockade of CD2 surface expression, maintaining immune cell proliferation and function, allowing for effective targeting of cancer cells without fratricide and improved therapeutic consistency.

Implementation Method 1

a CD2 blocking polypeptide comprising a single chain variable fragment (scFv) that binds CD2 linked to the N-terminus of a cellular localizing domain, wherein the cellular localizing domain comprises an amino acid sequence selected from the group consisting of an endoplasmic reticulum (ER) retention sequence, a Golgi retention sequence, and a proteosome localizing sequence, and wherein said CD2 blocking polypeptide remains intracellularly within said engineered cell and binds endogenous CD2 within the engineered cell

Methodology Applied

Scientific EffectIntracellular binding:

Implementation Method 2

the cellular localizing domain comprises an amino acid sequence selected from the group consisting of an endoplasmic reticulum (ER) retention sequence, a Golgi retention sequence, and a proteosome localizing sequence

Methodology Applied

Scientific EffectEndoplasmic reticulum retention:

Implementation Method 3

Methodology Applied

Scientific EffectGolgi retention:

Implementation Method 4

Chimeric antigen receptors (CARs) can redirect immune cells to specifically recognize and kill tumor cells. CARs are artificial multi-molecular proteins constituted by a single-chain variable region (scFv) of an antibody linked to a signaling molecule via a transmembrane domain. When the scFv ligates its cognate antigen, signal transduction is triggered

Methodology Applied

Scientific EffectAntigen recognition:

Implementation Method 5

When the scFv ligates its cognate antigen, signal transduction is triggered, resulting in tumor cell killing by CAR-expressing cytotoxic T lymphocytes

Methodology Applied

Scientific EffectSignal transduction:

Implementation Method 6

Chimeric antigen receptors (CARs) can redirect immune cells to specifically recognize and kill tumor cells... resulting in tumor cell killing by CAR-expressing cytotoxic T lymphocytes

Methodology Applied

Scientific EffectCytotoxicity:

Data Source

PatentUS12129485B2Blockade of CD2 surface expression and expression of chimeric antigen receptors for immunotherapy of T-cell malignancies

Publication Date: 2024.10.29 NATIONAL UNIVERSITY OF SINGAPORE

AI summary

The present invention provides engineered immune cells comprising an anti-CD2 protein expression blocker (PEBL) and an anti-CD2 chimeric antigen receptor (CAR). In some embodiments, such engineered immune cells lack surface expression CD2. Also, provided herein are methods of using such cells in cancer therapies.