STRC / DFNB16 — Problem Context for Research Routines

Agent context file. Every scheduled routine working in this vault reads this first to understand the current research state before acting. Mirrors ~/STRC/AGENTS.md philosophy and [[STRC Hypothesis Ranking]].

Last synced: 2026-04-24


Core problem

STRC encodes stereocilin — an extracellular protein that maintains the structure of the hair bundle on outer hair cells (OHCs) of the cochlea. Stereocilin forms the horizontal top connectors between stereocilia and attaches the tallest stereocilia to the tectorial membrane. Without it, hair bundles collapse, OHCs lose mechanosensitivity, and hearing fails.

Mutations in STRC cause DFNB16 — autosomal recessive non-syndromic hearing loss. Target patient: Misha (compound heterozygote — paternal 98 kb deletion / maternal c.4976A>C E1659A).

Central engineering constraint

STRC CDS is ~5,320 bp. AAV payload limit ~4,700 bp. STRC doesn’t fit a single AAV. Every mechanism hypothesis addresses this: truncate, split, edit in situ, use a non-AAV chassis, or replace function via materials.


Active hypothesis register

Source of truth: ~/Brain/notes/STRC Hypothesis Ranking.md. Full table below.

Scoring: M = Mechanism, D = Delivery, F = Misha-fit (each 1-5). Tier ≈ min(M, D, F). Tiers: S (primary, active compute, top-5 max) / A (active backburner) / B (watch) / C (paused) / D (killed) / reference (supporting models).

Canonical view. To change a row — edit the hub’s frontmatter (tier, mech, deliv, misha_fit, next_step), not this file. All 18 numbered hypotheses (h01–h16, h26, h27) are covered. Ref rows (supporting notes, clinical plans) are listed separately below.

TABLE WITHOUT ID
  hypothesis_num AS "#",
  link(hypothesis_title) AS "Hypothesis",
  mech AS "Mech",
  deliv AS "Deliv",
  misha_fit AS "Misha",
  tier AS "Tier",
  next_step AS "Next step",
  file.link AS "Hub"
FROM "hypotheses"
WHERE type = "hypothesis-hub"
SORT hypothesis_num ASC
#HypothesisMechDelivMishaTierNext stepHub
1STRC Pharmacochaperone Virtual Screen E1659A434AWetlab handoff packet v0.1 delivered 2026-04-27. H01 is ready for assay-scoping RFQ, not lead-candidate nomination. P0 next: send separate RFQs for (1) v5.3 custom synthesis of 1-indanyl_acylsulfonamide_SO2Me_-Cl + adamantyl_acylsulfonamide_SO2Me_-Cl, optional adamantyl_-CF3; (2) STRC 1066-1216 K1141-fragment WT/E1659A expression feasibility; (3) DSF/nanoDSF + SPR/BLI/MST direct-binding/stabilization cascade. Production holo MD/MM-PBSA remains paper-grade support, not wetlab-contact blocker. A held; mech 4 / deliv 3 / misha_fit 4 unchanged. A_hold_wetlab_rfq_feasibility. STRC h01 Wetlab Handoff Packet 2026-04-27index
10STRC In Situ SpyCatcher Assembly222CWet-lab only if #3 failsindex
11STRC Engineered TECTA Chimera222CAlt scaffold deferredindex
12Sonogenetic STRC Computational Proof222CSpeculative; no near-term pathindex
13STRC Programmable Recombinases213CTechnology-watch onlyindex
14STRC Protein Replacement Therapy213CNo delivery routeindex
15STRC OTOA Paralog Cross-Rescue222DChimera dead; upregulation weakindex
16STRC ZP Domain Prion-Like Seeding321DWrong patient (paternal 98kb Δ)index
2STRC Piezoelectric TM Bioelectronic Amplifier215CBlocked on literature — V_saturation, OHC membrane resistance, 210 kPa Young’s modulus all unsourced; TM displacement 5–30 nm is in-house assumption. No compute advance until primary lit retrieved.index
26STRC Engineered Homodimer Avidity254CPaused: Phase 1d A1078C/S1080C/S1579C/A1078W all fail AF3 G1; no further compute unless a new scaffold/interface appearsindex
27STRC STRCP1 Activation Rescue224CEmail Holt lab ribosome-profilingindex
3STRC Mini-STRC Single-Vector Hypothesis554SOrder Ultra-Mini gBlock + B8+WPRE3 cassette; clone pAAV B8-IgK-Ultra-Mini-WPRE3-bGH; Phase 4 HEK coIP. Verify B8 exact cloned sequence against Zhao 2025 Table S2 before GMP submission (cloning QC, not design blocker).index
4STRC mRNA-LNP Strategy B Full-Length322BNo fast compute-to-S path; keep as AAV maintenance/interim only unless new OHC-LNP tropism data reaches therapeutic population coverageindex
5STRC Calcium Oscillation Acoustic Therapy342CBlocked — 6 load-bearing parameters without primary lit (STRC mRNA t½, STRC protein t½, STRC copyindex
6STRC mRNA Therapy Hypothesis222BNo fast compute-to-S path; needs primary OHC RBM24 titration/Sun 2026 quantification before another model can change tierindex
7Prime Editing for STRC321CWet-lab ribosome-profiling STRCP1index
8STRC ASO Exon Skipping232CPhase 3a morpholino + 3b gapmerindex
9STRC Synthetic Peptide Hydrogel HTC543SReviewer-pack queue staged 2026-04-27 after Phase 4m S promotion. Next sessions should start at STRC h09 Reviewer Pack Queue 2026-04-27 and work in order: premortem precedent table orthogonal WH2 controls transfer memo. Phase 4m remains the gate-closing evidence (R2=0.690, Kd_F=1.48 mM); Phase 4k is a supporting caution (geometric contacts yes, confidence-supported contacts no). Phase 4l optional only if a reviewer needs paper-grade C_eff.index

Supporting notes (ref — no hub, not in active register)

These hypotheses/models feed into the active register above but don’t get their own hypothesis-hub folder. They’re either engineering sub-layers, survey notes, or clinical strategy docs.

Hypothesis / modelRole
STRC RBM24 Regulatory HypothesisRolled into Strategy A
STRC Stereocilia Bundle Mechanics ModelSupporting biophysics model
STRC AAV Vector DesignMini-STRC implementation detail
STRC B8 Enhancer SelectionAAV pipeline
STRC Dual-Vector vs Single-Vector TransductionEngineering layer
STRC Anti-AAV Immune Response ModelRe-dosing constraint
STRC Electrostatic Analysis E1659APharmacochaperone input
Adult Treatment Window STRCConstraint on all mechanisms
Alternative STRC Delivery HypothesesDelivery-layer survey
Misha Compound-Het Therapy Stack ModelClinical plan: parallel h01+h03 stack

All 18 numbered hypotheses (h01–h16, h26, h27) now live in hypotheses/h{N}/hub.md and render via the Dataview block above. Manual table removed 2026-04-23 — edit hub frontmatter to change tier/score/next-step.


Hypothesis → synthesis file slug

When Part 2 (hypothesis synthesis) needs to write a Recent Papers entry, use these slugs. null = no synthesis file yet; routine should skip synthesis for that hypothesis.

HypothesisSynthesis file
STRC Pharmacochaperone Virtual Screen E1659Anull
STRC Piezoelectric TM Bioelectronic Amplifiernull
STRC Mini-STRC Single-Vector Hypothesishypotheses/mini-strc.md
STRC mRNA-LNP Strategy B Full-Lengthnull
STRC Calcium Oscillation Acoustic Therapynull
STRC mRNA Therapy Hypothesisnull
Prime Editing for STRChypotheses/prime-editing.md
STRC ASO Exon Skippinghypotheses/exon-skipping.md
STRC Synthetic Peptide Hydrogel HTCnull
STRC In Situ SpyCatcher Assemblynull
STRC Engineered TECTA Chimeranull
Sonogenetic STRC Computational Proofhypotheses/sonogenetics.md
STRC Programmable Recombinaseshypotheses/recombinases.md
STRC Protein Replacement Therapynull
STRC OTOA Paralog Cross-Rescuenull
STRC ZP Domain Prion-Like Seedingnull
STRC RBM24 Regulatory Hypothesisnull
STRC Stereocilia Bundle Mechanics Modelhypotheses/bundle-mechanics.md
STRC AAV Vector Designnull
STRC B8 Enhancer Selectionhypotheses/enhancer.md
STRC Dual-Vector vs Single-Vector Transductionhypotheses/dual-vector.md
STRC Anti-AAV Immune Response Modelhypotheses/immune.md
STRC Electrostatic Analysis E1659Ahypotheses/electrostatics.md
Adult Treatment Window STRCnull
Alternative STRC Delivery Hypotheseshypotheses/delivery.md

Kill list (D-tier — do not pursue)

TABLE WITHOUT ID
  link(hypothesis_title) AS "Hypothesis",
  next_step AS "Reason killed",
  file.link AS "Hub"
FROM "hypotheses"
WHERE type = "hypothesis-hub" AND tier = "D"
SORT hypothesis_num ASC
HypothesisReason killedHub
STRC OTOA Paralog Cross-RescueChimera dead; upregulation weakindex
STRC ZP Domain Prion-Like SeedingWrong patient (paternal 98kb Δ)index

How to use this file (agent directive)

  1. Before scanning: read this entire file. Understand S-tier (top compute), A-tier (backburner), killed. Skew relevance scoring toward S/A — papers advancing S-tier hypotheses are HIGH value. Papers on D-tier should only be indexed if they credibly flip the kill.
  2. For paper dedup: see routine Step 1 (builds DOI/PMID/bioRxiv lookup sets).
  3. For hypothesis synthesis (Part 2): match papers to hypotheses by tag + relevance + mechanism overlap. Append “Recent Papers” entries only to files listed in the slug table. If null: log SKIP-NO-SYNTH-FILE: <hypothesis> and move on.
  4. If this file feels stale (S-tier hypothesis with completed next-step, or paper mentions hypothesis not in the table): operate from this file anyway, flag drift in commit message.

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