CRISPR Epitope Engineering for Antigen-Sparing Hematopoietic Cells
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Solution Overview
Problem
Immunotherapies targeting specific antigens can deplete both pathological and non-pathological cells, leading to severe side effects due to 'on-target, off-disease' effects, particularly when the targeted antigen is essential for cell survival.
Innovation Solution
Genetically modify hematopoietic cells to alter the amino acid sequence of lineage-specific cell-surface antigens using CRISPR/Cas systems combined with homology-directed repair (HDR), reducing the binding of immunotherapeutic agents without impairing the antigen's function.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If immunotherapy targets a specific antigen to treat disease, then therapeutic effectiveness is improved, but harmful off-disease effects increase due to depletion of non-pathological cells
Solution Approach 1:
The patent applies local quality by modifying the epitope (local region) of the cell-surface antigen while preserving the rest of the protein's structure and function. This allows the antigen to maintain its role in cell survival while the modified epitope prevents immunotherapeutic binding, thus resolving the contradiction between therapeutic effectiveness and harmful off-disease effects
Solution Approach 2:
The modified epitope acts as an intermediary element that mediates between the need for antigen presence (for cell survival) and the need to prevent immunotherapeutic binding. The epitope modification serves as a protective mechanism that allows cells to express the antigen without being targeted by CAR-T cells or other immunotherapies
2Reliability
If the targeted antigen is essential for cell survival, then cell viability is improved, but the subject cannot receive immunotherapy or faces severe side effects
Solution Approach 1:
The patent modifies only the local epitope region of the antigen while preserving the overall protein structure and its essential functions for cell survival. This localized modification allows the antigen to continue supporting cell viability while preventing recognition by immunotherapeutic agents, thereby eliminating severe side effects
Solution Approach 2:
The epitope modification is performed in advance on the cell-surface antigen before immunotherapy administration. This preliminary action ensures that when the immunotherapy is administered, the modified antigen already prevents binding to CAR-T cells or other effector cells, preventing severe side effects before they can occur
3Object-affected harmful factors
If genetic modification is used to alter the epitope, then agent binding is reduced, but manufacturing complexity increases
Solution Approach 1:
The patent uses homology-directed repair (HDR) with a template DNA sequence that copies the desired epitope modification into the cell's genome. This copying mechanism allows precise epitope alteration through a well-established molecular biology process, managing manufacturing complexity by using a proven and controllable method
Solution Approach 2:
The patent changes the nucleotide sequence parameters of the epitope region through HDR, transforming the DNA sequence to encode the modified epitope. This parameter change approach allows precise control over the epitope modification while using standard genetic engineering tools, thereby managing manufacturing complexity through systematic and controllable modifications
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
Achieves high editing efficiency and viability in edited cells, mitigating detrimental on-target, off-disease effects by minimizing agent binding to modified antigens, thus preserving cell functionality and reducing side effects.
Implementation Method 1
contacting the cell with a CRISPR/Cas system, wherein the contacting creates a double-stranded break in a sequence of the lineage-specific cell-surface antigen
Implementation Method 2
contacting the cell with a template polynucleotide, wherein the contacting results in integration of a sequence of the template polynucleotide at the site of the double-stranded break through homology-directed repair
Data Source
AI summary
Provided herein are compositions and methods for genetically engineering a cell (e.g., a hematopoietic cell) to modify a gene encoding a lineage-specific cell-surface antigen to modify an epitope of the lineage-specific cell-surface antigen recognized by an agent. Also provided are methods involving administering such genetically engineered cells to a subject, such as a subject having a hematopoietic malignancy, as well as the genetically engineered cells themselves.