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Part 2

Research Hypotheses

Computational hypotheses for accelerating STRC gene therapy. These require experimental validation.

AAV packaging

Mini-STRC Hypothesis

Three single-AAV constructs validated by 16 AF3 experiments. Removing the disordered N-terminal improves folding (pTM 0.81-0.87). Recommended: shorter mini-STRC (3228 bp, 1472 bp headroom).

In situ correction

Prime Editing

Precision correction of c.4976A>C using PE3 or PE3b pegRNA strategy without double-strand breaks.

Non-invasive

Sonogenetics

Mechanosensitive channel activation via focused ultrasound. ODE model of calcium signaling in OHC stereocilia.

Sonoporation

Delivery

Non-invasive cochlear gene delivery via sonoporation + LNP. ODE model shows 78% transduction efficiency under optimized parameters.

Stochastic model

Dual-Vector vs Single-Vector

Poisson stochastic model quantifying the probability cost of dual-vector co-transduction vs single-vector strategy.

Biophysics

Electrostatic Analysis

Coulomb energetics explain why E1659A is pathogenic despite intact structure. 8.62 kcal/mol destabilization of adhesion interface.

Pharmacokinetics

Immune Response

NAb kinetics, seroprevalence barriers, and delivery vehicle comparison for cochlear gene therapy re-dosing strategies.

Cell targeting

Enhancer-Driven OHC Targeting

ARBITER workflow for OHC-specific synthetic enhancers. B8 enhancer fully rescues hearing with zero ectopic expression. Potential synergy with mini-STRC for single-vector therapy.

RNA therapeutics

ASO Exon Skipping

Antisense oligonucleotides to skip mutant exons, producing shorter but functional stereocilin. Repeatable, no AAV needed. Proven in DMD (4 FDA-approved drugs).