ADAR-Specific Guide RNAs for Tumor Microenvironment Modulation
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Solution Overview
Problem
Current cancer therapies face challenges in effectively modulating the tumor microenvironment to enhance immunogenicity, as cancer cells often evade immune checkpoints, and there is a need for specific gene targets to improve anti-tumor immune responses.
Innovation Solution
The development of DNA targeting systems using guide RNAs (gRNAs) to modify the ADAR gene, specifically reducing its expression in mammalian cells, which can modulate the tumor microenvironment and enhance immune responses by administering a site-directed modifying polypeptide and gRNA combination, potentially combined with immune checkpoint blockade agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If immune checkpoint blockade therapy is used to treat cancer, then immune response against tumor is enhanced, but cancer cells can evade immune checkpoints reducing treatment effectiveness
Solution Approach 1:
The patent extracts and targets the specific gene ADAR as a new therapeutic target beyond conventional immune checkpoints. By using CRISPR-Cas9 to specifically knock out the ADAR gene in cancer cells, the invention removes the evasion mechanism that allows cancer cells to escape immune detection, thereby enhancing the reliability of immunomodulatory treatment.
Solution Approach 2:
The invention changes the therapeutic parameter from targeting immune checkpoint molecules to targeting the ADAR gene product. This parameter change involves using guide RNAs with specific sequences complementary to ADAR gene regions, fundamentally altering the approach to cancer immunotherapy by addressing a different molecular target that cancer cells exploit for immune evasion.
2Adaptability or versatility
If CRISPR systems are used to target genes for cancer treatment, then gene modification capability is achieved, but identification of effective gene targets remains challenging
Solution Approach 1:
The patent performs preliminary action by identifying and validating the ADAR gene as an effective target before clinical application. The invention provides specific guide RNA sequences that have been pre-tested to successfully target and knock out the ADAR gene, eliminating the need for de novo target identification and simplifying the implementation of CRISPR-based cancer therapy.
Solution Approach 2:
The invention creates copies of guide RNA sequences with high sequence identity to the ADAR gene target regions. These replicated guide sequences ensure consistent and reliable targeting of the ADAR gene across different experimental and clinical applications, making the gene targeting process more predictable and easier to implement.
3Productivity
If ADAR gene expression is reduced to modulate tumor microenvironment, then tumor growth is reduced, but specific delivery and targeting efficiency must be maintained
Solution Approach 1:
The patent segments the therapeutic approach into distinct functional components: guide RNAs with specific sequences (e.g., SEQ ID NOs: 139-148, 151-194) that precisely recognize ADAR gene regions, and the CRISPR-Cas9 machinery. This segmentation allows for optimized delivery of each component and ensures precise targeting of the ADAR gene to achieve tumor growth reduction while maintaining delivery efficiency.
Data Source
AI summary
Provided herein are DNA targeting systems comprising a site-directed modifying polypeptide an ADAR-specific guide RNAs, and uses thereof.