CIITA Gene Knockout in Engineered Immune Cells for Universal CAR-T
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Solution Overview
Problem
Current CAR-T cell therapies face challenges such as graft-versus-host disease (GvHD) and host-versus-graft disease (HvGD) due to the proliferation of engineered cells attacking normal tissues or being rejected by the patient's immune system, limiting the development of universal CAR-T cells for allogeneic transplantation.
Innovation Solution
The use of an sgRNA specifically targeting the CIITA gene to knock out CIITA in immune cells, combined with other gene silencing, to create engineered immune cells that reduce immune rejection and GvHD, enabling the production of universal CAR-T cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If T cells are used for universal CAR-T production, then the vitality and function are superior and CAR infection rate is improved, but the patient's immune system may reject allogeneic CAR-T cells causing host-versus-graft disease
Solution Approach 1:
The patent extracts and removes the CIITA gene from allogeneic T cells using CRISPR/Cas9 technology. By specifically knocking out this gene that controls MHC class II expression, the invention eliminates the target antigen that would trigger host-versus-graft disease, thereby resolving the contradiction between using vigorous allogeneic T cells and avoiding immune rejection
Solution Approach 2:
The patent changes the genetic parameter of the T cells by knocking out the CIITA gene. This genetic modification alters the expression profile of MHC molecules, specifically reducing MHC class II expression to evade host immune recognition while preserving the functional vitality of the allogeneic T cells
2Productivity
If engineered CAR-T cells proliferate in the patient's body, then therapeutic effect is enhanced, but they may attack normal cells or tissues causing graft-versus-host disease
Solution Approach 1:
The patent removes the CIITA gene which is responsible for controlling MHC class II expression. By extracting this specific genetic element, the engineered CAR-T cells lose the ability to present antigens that would trigger graft-versus-host disease against normal host tissues, while retaining their proliferative and therapeutic capabilities
Solution Approach 2:
The patent converts the potential harm of CIITA-mediated MHC class II expression (which causes GvHD) into a benefit by knocking it out. The loss of CIITA function, which would normally be detrimental to immune recognition, is actually beneficial in this context because it prevents the engineered cells from attacking host tissues while maintaining their anti-tumor activity
3Reliability
If multiple genes are knocked out to reduce transplantation risk, then GvHD and HvGD are reduced, but the device complexity and manufacturing difficulty increase
Solution Approach 1:
The patent focuses on extracting and knocking out specifically the CIITA gene rather than multiple genes. This targeted approach simplifies the gene editing process compared to knocking out multiple genes simultaneously, while still achieving the goal of reducing both GvHD and HvGD risks through selective MHC class II suppression
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 CIITA knock-out strategy effectively reduces immune rejection and GvHD, allowing for the development of universal CAR-T cells with enhanced therapeutic efficacy and safety for allogeneic transplantation.
Implementation Method 1
The present disclosure provides an sgRNA and uses the sgRNA to knock out the CIITA gene to obtain an engineered immune cell
Data Source
AI summary
Disclosed herein are an engineered immune cell with CIITA gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CIITA gene is designed and synthesized, which can accurately target CIITA gene to achieve gene knock-out with high knockout efficiency. The provided sgRNA can be used for preparing a universal CAR-T cell.