CRISPR/Cas9 Vector Knockout for Xenotransplant Heart Valves
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Solution Overview
Problem
Current cardiac replacement therapies using mechanical artificial heart valves or glutaraldehyde-fixed wild-type heart valves from pig or bovine tissue face issues such as inactivation of vascular endothelial cells, calcification leading to structural valve degeneration, and significant immune rejection reactions, particularly in young patients, resulting in short-term transplantation success and increased morbidity and mortality.
Innovation Solution
A CRISPR/Cas9 vector system targeting and knocking out the GGTA1, CMAH, and β4GalNT2 genes in porcine tissues, which are primary antigens causing immune rejection, to reduce immunogenicity and prevent calcification, thereby enhancing the longevity and success of xenotransplanted heart valves.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If glutaraldehyde-fixed wild-type heart valves are used for cardiac replacement therapy, then the valve can be implanted clinically, but the vascular endothelial cells are inactivated and the transplantation effect is damaged
Solution Approach 1:
The patent extracts and removes the harmful antigens (GGTA1, CMAH, β4GalNT2) from the porcine heart valve tissue through CRISPR/Cas9-mediated gene knockout, eliminating the cause of immune rejection while preserving the valve's structural and functional properties for clinical transplantation
Solution Approach 2:
The patent changes the genetic parameters of the porcine heart valve by knocking out specific genes (GGTA1, CMAH, β4GalNT2) that encode immunogenic antigens, thereby altering the immunological properties of the valve to reduce immune rejection and improve transplantation success
2Duration of action of stationary object
If glutaraldehyde-fixed wild-type heart valves are used, then the valve can be implanted, but calcification occurs causing structural valve degeneration and requiring further operations
Solution Approach 1:
The patent converts the naturally occurring immunogenic antigens in porcine heart valves into a benefit by selectively removing them through gene knockout, thereby preventing the harmful cascade of immune rejection, calcification, and structural degeneration that would otherwise occur, extending valve durability
Solution Approach 2:
By altering the genetic composition of the porcine heart valve through CRISPR/Cas9-mediated knockout of GGTA1, CMAH, and β4GalNT2 genes, the patent changes the biochemical parameters of the valve tissue to prevent calcification and structural degeneration, thereby extending the functional lifespan of the implanted valve
3Ease of manufacture
If wild-type porcine heart valves are used for xenotransplantation, then the valve material is available, but strong immune rejection reactions occur particularly in young patients
Solution Approach 1:
The patent extracts and eliminates the specific immunogenic antigens (GGTA1, CMAH, β4GalNT2) from the porcine heart valve through targeted gene knockout, removing the cause of immune rejection while maintaining the availability of porcine heart valve material for xenotransplantation
Solution Approach 2:
The patent changes the immunological parameters of the porcine heart valve by knocking out genes encoding alpha-gal, Neu5Gc, and other xenogenic antigens, thereby reducing the immunogenicity of the valve material and minimizing immune rejection reactions in xenotransplantation recipients
4Reliability
If young patients receive GBHV transplantation, then the valve can be implanted, but the transplantation success rate is low due to stronger immune system and reject reaction
Solution Approach 1:
The patent changes the genetic and immunological parameters of the porcine heart valve by knocking out multiple antigen-encoding genes simultaneously, creating a hypoimmunogenic valve that can withstand the strong immune response of young patients and achieve sustained transplantation success
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 CRISPR/Cas9-mediated knockout of GGTA1, CMAH, and β4GalNT2 genes significantly decreases the immune rejection reaction and calcification, leading to a longer-lasting and more effective xenotransplanted heart valve solution, addressing the limitations of existing cardiac replacement therapies.
Implementation Method 1
A CRISPR/Cas9 vector system targeting and knocking out the GGTA1, CMAH, and β4GalNT2 genes in porcine tissues
Data Source
AI summary
Provided is an SgRNA combination, comprising an SgRNA specifically targeting the GGTA1 gene, an SgRNA specifically targeting the CMAH gene and an SgRNA specifically targeting the β4GalNT2 gene. Also provided is a CRISPR/Cas9 vector combination, comprising a GGTA1-CRISPR/Cas9 vector, a CMAH-CRISPR/Cas9 vector and a β4GalNT2-CRISPR/Cas9 vector. Also provided is an applicaton of the CRISPR/Cas9 vector combination in knocking out the GGTA1 gene, the CMAH gene and the β4GalNT2 gene. The knockout rates of the three genes with the specifically targeted SgRNA sequences are respectively 56%, 63%, and 41%. A three genes knockoutpig can be obtained, wherein the three genes related to immune rejectionare knocked out, and heart valves of said pig can be acquired.