B2M Gene Editing in B Cells for Allogeneic Transplantation
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Solution Overview
Problem
Current genome editing techniques face challenges in clinically relevant human somatic cells due to unwanted host immune responses during allogeneic transplantation, necessitating cells that minimize immune rejection.
Innovation Solution
The method involves modifying the endogenous beta-2 microglobulin (B2M) gene in B cells using a nuclease capable of cleaving the targeted locus, accompanied by guide RNA and a repair template, to inactivate the B2M gene or insert a replacement MHC-I, thereby increasing resistance to allogeneic immune cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If genome editing is performed in human somatic cells, then the ability to express therapeutic proteins is improved, but unwanted host immune responses occur during allogeneic transplantation
Solution Approach 1:
The patent removes the endogenous B2M gene from human somatic cells using CRISPR-Cas9 genome editing technology. By extracting this specific gene component, the cells lose the ability to express MHC class I molecules, thereby eliminating the target for allogeneic T cell recognition and reducing immune rejection during transplantation while preserving therapeutic protein expression capability
Solution Approach 2:
The patent changes the genetic parameter of the B2M gene by creating a knockout mutation. This parameter change fundamentally alters the cell's MHC class I expression status, transforming it from MHC-I positive (immunogenic) to MHC-I negative (immune-evasive), thereby resolving the contradiction between therapeutic utility and immune compatibility
2Reliability
If the endogenous B2M gene is inactivated, then resistance to killing by allogeneic immune cells is increased, but the cell may become susceptible to other immune mechanisms
Solution Approach 1:
The patent converts the potential harm of B2M inactivation (susceptibility to NK cells) into a benefit by simultaneously eliminating MHC class I expression, which provides greater protection against the more potent threat of allogeneic T cell-mediated cytotoxicity. The net effect is improved survival in allogeneic transplantation contexts
Solution Approach 2:
The patent changes the immunophenotypic parameter of the cell by knocking out B2M, fundamentally altering how the cell interacts with the allogeneic immune system. This parameter change shifts the balance of immune interactions in favor of cell survival despite potential NK cell susceptibility
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 enhances the resistance of modified B cells to killing by allogeneic immune cells, allowing for successful allogeneic transplantation by reducing immune rejection and enabling the expression of therapeutic proteins.
Implementation Method 1
a nuclease capable of cleaving a targeted locus in an endogenous B2M gene in a genome of the cell
Implementation Method 2
a guide RNA (gRNA) comprising a sequence complementary to the B2M gene
Implementation Method 3
a repair template comprising a first homology arm, a second homology arm, and a nucleic acid encoding a payload therebetween, wherein the first homology arm and/or the second homology arm has homology to a sequence in the B2M gene
Data Source
AI summary
Some embodiments of the methods and compositions provided herein include preparing modified B cells. In some embodiments, an endogenous beta-2 microglobulin (B2M) gene in a B cell is modified. Some embodiments relate to increasing the resistance of modified B cells to killing by allogeneic immune cells. In some embodiments, the endogenous B2M gene is inactivated increasing the resistance of the modified B cell to killing by allogeneic immune cells. In some embodiments, a replacement MHC-I is inserted into an inactivated endogenous B2M gene increasing the resistance of the modified B cell to killing by allogeneic immune cells. Some embodiments include enriching for successfully modified cells.