CRISPR CD7 Knockout for CAR-T Fratricide Prevention
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Solution Overview
Problem
Current immunotherapies, such as CAR-T therapies, often cause 'on-target, off-disease' effects where healthy cells expressing the targeted antigen are depleted, leading to severe side effects or rendering the therapy ineffective due to the shared expression of antigens on both malignant and healthy cells.
Innovation Solution
Genetically engineered cells with reduced or absent expression of CD7, achieved through CRISPR/Cas-mediated genome editing, are developed to avoid recognition and targeting by immunotherapies, allowing them to engraft and differentiate in the bone marrow without being affected by CD7-specific immunotherapies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If immunotherapy targets an antigen expressed on both malignant and healthy cells, then the therapy can effectively target and destroy malignant cells, but it causes depletion of healthy cells leading to severe side effects
Solution Approach 1:
The patent extracts the harmful antigen expression from healthy cells by using CRISPR/Cas9 genome editing to knock out the targeted antigen (e.g., CD7) in healthy hematopoietic cells. This removes the target from healthy cells while leaving it intact in malignant cells, allowing immunotherapy to selectively target only the malignant cells without causing harmful effects on healthy cells.
Solution Approach 2:
The patent applies preliminary action by pre-modifying healthy hematopoietic cells with CRISPR/Cas9 to knock out the targeted antigen before administering the immunotherapy. This preliminary genetic modification ensures that when the immunotherapy is administered, healthy cells do not express the target antigen and thus are not affected, while malignant cells remain susceptible to targeting.
2Adaptability or versatility
If immunotherapy targets an antigen expressed on immune effector cells, then the therapy can be designed to be specific, but it causes fratricide of the effector cells rendering the therapy ineffective
Solution Approach 1:
The patent removes the targeted antigen from healthy immune effector cells through CRISPR/Cas9-mediated gene knockout. This extraction ensures that when CAR-T cells are generated with specificity for that antigen, they will not recognize and destroy their own healthy progeny, thereby preventing fratricide and maintaining therapy effectiveness.
Solution Approach 2:
The patent applies preliminary action by performing the antigen knockout in healthy hematopoietic stem cells or progenitors before they differentiate into functional immune effector cells. This ensures that the antigen is absent from all downstream effector cells, including CAR-T cells, preventing self-targeting and fratricide while allowing the therapy to remain specific for malignant cells.
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 genetically modified cells effectively engraft and reconstitute hematopoietic lineages, reducing on-target, off-disease cytotoxicity and enhancing the therapeutic index of immunotherapies by minimizing harm to healthy cells while maintaining efficacy against malignant cells.
Implementation Method 1
Genetically engineered cells with reduced or absent expression of CD7, achieved through CRISPR/Cas-mediated genome editing
Implementation Method 2
contacting the cell with (i) any of the gRNAs described herein or a gRNA targeting a targeting domain targeted by any of the gRNAs described herein; and (ii) an RNA-guided nuclease that binds the gRNA, thus forming a ribonucleoprotein (RNP) complex
Implementation Method 3
allowing them to engraft and differentiate in the bone marrow without being affected by CD7-specific immunotherapies
Data Source
AI summary
Provided herein are gRNA comprising a targeting domain that targets CD7, which may be used, for example, to make modifications in cells. Also provided herein are methods of genetically engineered cell having a modification (e.g., insertion or deletion) in the CD7 gene and methods involving administering such genetically engineered cells to a subject, such as a subject having a hematopoietic malignancy.