IL6R scFv CAR-T Vector for CRS Alleviation
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Solution Overview
Problem
Current CAR-T cell therapy for cancer treatment often results in cytokine release syndrome (CRS), which is costly to manage with drugs like tocilizumab and can inhibit the therapeutic effect of CAR-T cells, necessitating a more cost-effective and efficient method to block IL6R signaling without compromising the efficacy of CAR-T cell function.
Innovation Solution
A CAR-T transgenic vector is developed using a lentiviral vector system that expresses a human IL6R single-chain antibody (scFv) to block IL6R signaling, utilizing a recombinant lentiviral vector design that includes specific promoters, signal peptides, and a four-plasmid packaging system for efficient transduction and expression, thereby reducing the need for expensive antibody drugs and enhancing the therapeutic effect of CAR-T cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If commercial antibody drugs like tocilizumab are used to block IL6R signaling, then CRS symptoms can be alleviated, but the cost of treatment increases significantly
Solution Approach 1:
The CAR-T cells are genetically modified to self-produce the scFv protein that blocks IL6R signaling. The cells autonomously synthesize and secrete the blocking antibody fragment, eliminating the need for external antibody drug administration and reducing treatment costs while maintaining CRS alleviation efficacy
Solution Approach 2:
Instead of using expensive commercial antibody drugs, the invention creates a copy of the therapeutic function by integrating the scFv gene into the CAR-T cell genome. The cells then produce their own blocking agent, replicating the therapeutic effect of tocilizumab but through an endogenous, cost-effective mechanism
2Object-affected harmful factors
If anti-histamine drugs or corticosteroids are used to inhibit inflammatory response, then CRS symptoms are reduced, but the killing effect of CAR-T cells on tumors is inhibited
Solution Approach 1:
The invention applies localized, targeted blocking of IL6R signaling specifically at the site of cytokine action. The scFv protein is secreted by CAR-T cells to locally neutralize IL-6, providing precise inhibition of the harmful inflammatory pathway without broad suppression of immune function that would compromise tumor killing
Solution Approach 2:
The scFv protein acts as an intermediary that specifically binds to IL-6, preventing it from activating the IL6R-JAK-STAT signaling pathway. This intermediate blocking mechanism selectively interrupts the harmful cytokine signal while preserving the functional integrity of CAR-T cells for tumor targeting
3Object-affected harmful factors
If expensive commercial antibody drugs are used to block IL6R, then CRS is treated, but the therapeutic efficiency and cost-effectiveness are compromised
Solution Approach 1:
CAR-T cells are engineered to autonomously produce and secrete the scFv blocking protein, eliminating dependence on external antibody drug administration. This self-sufficient approach ensures continuous local blocking of IL6R signaling, improving therapeutic efficiency while eliminating the high costs of commercial antibody drugs
Solution Approach 2:
The invention merges the CAR-T cell therapy function with the IL6R blocking function into a single cellular platform. The dual functionality is achieved by co-expressing the CAR receptor and scFv gene in the same cells, creating an integrated therapeutic agent that simultaneously provides tumor targeting and CRS prevention
Data Source
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AI summary
The invention discloses a IL6R block CAR-T transgenic vector for alleviating CRS, including: AmpR sequence containing ampicillin resistance gene (SEQ ID NO.1); prokaryotic replicon pUC Ori sequence (SEQ ID NO.2); virus replicon SV40 Ori sequence (SEQ ID NO.3); eWPRE enhanced posttranscriptional regulatory element of hepatitis B virus (SEQ ID NO.11); human EFla promoter (SEQ ID NO.12); lentiviral packaging cis-elements for lentiviral packaging; humanized single-chain antibody fragment IL6RscFv1 (SEQ ID NO.21) , IL6RscFv2 (SEQ ID NO.22), or IL6RscFv3 (SEQ ID NO.23) of human IL6R; IRES ribosome binding sequence (SEQ ID NO.25); IL6 signal peptide (SEQ ID NO.26); human antibody Fc segment (SEQ ID NO.27); and the chimeric antigen receptors of the second or third generation CAR for integrating recognition, transmission and initiation. Also, the invention discloses the preparation method of the vector and its application in the preparation of drugs alleviating CRS.