Mutant ER-LBD Switches for Selective Synthetic-Ligand Cell Killing
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Solution Overview
Problem
Existing inducible cell death systems based on estrogen receptor ligand binding domains (ER-LBD) lack sufficient sensitivity and selectivity to synthetic ligands, posing challenges in therapeutic applications such as cell and gene therapy where precise control over cell killing is required.
Innovation Solution
Development of a modified ER-LBD with specific amino acid substitutions, including G400V, M543A, L544A, and optionally V595A, along with additional substitutions at specific positions, enhancing sensitivity and selectivity to non-endogenous ligands, thereby improving the inducible cell death system's responsiveness and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a modified ER-LBD with specific amino acid substitutions is developed, then sensitivity and selectivity to synthetic ligands is improved, but device complexity increases
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid substitutions at specific positions (G400V, M543A, L544A, V595A) in the ER-LBD to optimize ligand binding characteristics. This resolves the contradiction by achieving enhanced sensitivity and selectivity through controlled parameter modification rather than fundamental structural changes.
Solution Approach 2:
The invention implements local quality by introducing specific amino acid substitutions at targeted positions within the ligand binding domain while maintaining the overall structural integrity of the polypeptide. This localized modification approach improves ligand interaction properties without requiring comprehensive restructuring of the entire system.
2Measurement precision
If sensitivity to non-endogenous ligands is increased, then control precision over cell death induction is improved, but selectivity between endogenous and non-endogenous ligands may be compromised
Solution Approach 1:
The patent resolves this contradiction by carefully selecting specific amino acid substitutions (G400V, M543A, L544A, V595A) that shift the binding parameter profile toward non-endogenous ligands while maintaining discrimination against endogenous ligands. The modifications are designed to achieve optimal binding affinity for synthetic ligands without compromising overall selectivity.
Solution Approach 2:
The invention utilizes the natural ER-LBD structure as a template and creates modified versions through targeted amino acid substitutions. This copying approach allows optimization of ligand binding properties while preserving the fundamental selectivity mechanisms of the original receptor structure.
Data Source
AI summary
Provided herein are mutants of estrogen receptor alpha ligand binding domain (ER-LBD), and inducible cell death systems that include mutants of estrogen receptor alpha ligand binding domain (ER-LBD). Also provided are methods of for use of the same, such as inducing cell death in a cell.