Universal Donor Cells With MHC Editing for Broad Immune Evasion
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for generating universal donor cells face challenges in effectively evading immune rejection and ensuring cell survival post-engraftment, with potential issues like residual MHC-I expression and susceptibility to natural killer cells, and there is a need for improved strategies to generate cells that can evade immune responses and survive in host environments.
Innovation Solution
Genetic modifications are introduced in cells to delete or modify MHC-I and MHC-II genes and introduce tolerogenic factors like PD-L1 and HLA-E, using CRISPR systems for precise editing, combined with survival factor enhancements to improve immune evasion and cell survival.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If MHC-I and MHC-II genes are disrupted to reduce immunogenicity, then immune rejection is reduced, but residual MHC-I expression may still occur causing immune rejection
Solution Approach 1:
The patent uses CRISPR-Cas9 genome editing to precisely disrupt MHC class I and class II genes, achieving complete gene knockout rather than partial disruption. This parameter change from partial to complete gene disruption ensures no residual MHC-I expression occurs, resolving the contradiction between reducing immunogenicity and ensuring complete gene disruption.
Solution Approach 2:
The patent replaces traditional mechanical or chemical gene disruption methods with CRISPR-Cas9 genome editing technology. This substitution enables precise, targeted, and complete gene disruption of MHC class I and class II genes, eliminating residual expression issues that plague conventional approaches.
2Reliability
If MHC class I genes are depleted to evade CD8+ T cells, then allogeneic rejection is reduced, but cells become susceptible to natural killer cell lysis
Solution Approach 1:
The patent converts the harmful effect of MHC class I depletion (NK cell susceptibility) into a benefit by simultaneously depleting MHC class II genes and introducing tolerogenic factors. This dual approach ensures that while cells are vulnerable to NK cells, they are protected from both CD8+ and CD4+ T cell responses, creating an overall immune evasion advantage.
Solution Approach 2:
The patent creates a composite genetic modification strategy combining MHC class I and class II gene disruption with tolerogenic factor introduction. This composite approach addresses multiple immune rejection pathways simultaneously, balancing NK cell susceptibility with profound protection against T cell-mediated rejection.
3Reliability
If multiple genetic modifications are introduced to achieve universal donor status, then immune compatibility is improved, but cell complexity and manufacturing difficulty increase
Solution Approach 1:
The patent achieves universal donor status by simultaneously targeting both MHC class I and class II pathways with CRISPR-Cas9, creating cells that are broadly compatible across different HLA genotypes. This multi-functional approach consolidates multiple immune evasion mechanisms into a single universal cell product.
Solution Approach 2:
The patent replaces complex, multi-step traditional gene targeting methods with CRISPR-Cas9 genome editing, which simplifies the delivery and integration of multiple genetic modifications. This substitution reduces manufacturing complexity while achieving comprehensive MHC class I and class II disruption combined with tolerogenic factor introduction.
Data Source
AI summary
Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.