Double Knockout NK Cells CISH NKG2A
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Solution Overview
Problem
Current immune cell therapies, such as those using natural killer (NK) cells, face challenges in effectively targeting and killing cancer cells due to regulatory mechanisms like CISH and NKG2A, which limit cytotoxicity and persistence, necessitating enhanced methods for improved cancer treatment.
Innovation Solution
Modified NK cells and cell lines with reduced CISH and NKG2A function, optionally expressing TRAIL ligands, chimeric antigen receptors, and growth-promoting cytokines, are engineered to increase cytotoxicity and persistence, thereby enhancing their ability to target and kill cancer cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If NK cells are used to target cancer cells, then cancer cell killing ability is improved, but cytotoxic activity is dampened by inhibitory receptors like NKG2A and CISH
Solution Approach 1:
The patent extracts and removes the inhibitory elements (NKG2A receptor and CISH gene) from the NK cell system. By knocking out NKG2A and CISH, the harmful inhibitory signals that dampen cytotoxic activity are eliminated, allowing NK cells to maintain high cytotoxicity against cancer cells without being suppressed by these regulatory mechanisms
Solution Approach 2:
The patent changes the functional parameters of NK cells by modifying gene expression levels. Specifically, it reduces or eliminates the expression of inhibitory components (NKG2A and CISH), thereby altering the balance between inhibitory and activating signals to enhance cytotoxic activity against cancer cells
2Reliability
If cytokine concentrations are kept low to maintain regulatory control, then cell expansion is limited and cytotoxic activity decreases
Solution Approach 1:
The patent converts the harmful effect of CISH (which normally limits cell expansion and cytotoxicity under low cytokine conditions) into a benefit by knocking out the CISH gene. This eliminates the negative regulation, allowing NK cells to expand and maintain high cytotoxic activity even at lower cytokine concentrations, thus improving productivity without sacrificing control
3Object-affected harmful factors
If NK cells are modified to enhance cytotoxicity, then cancer cell killing is improved, but transfection efficiency and ease of clinical application may be compromised
Solution Approach 1:
The patent uses CRISPR/Cas9 gene editing to knock out inhibitory genes (NKG2A and CISH) directly in NK cells, rather than using traditional transfection methods to add complex transgenic constructs. This approach simplifies the manufacturing process, improves transfection efficiency, and makes clinical application more feasible while still achieving enhanced cytotoxicity
Data Source
Figure 1A~1B
Figure 2A~2B
AI summary
NK cells and NK cell lines are modified to increase their cytotoxicity, proliferation, metabolic profile and persistence, wherein the cells and compositions thereof have a use in the treatment of cancer. Production of modified NK cells and NK cell lines is via genetic modification to knockout expression of both CISH and NKG2A genes