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VDAC1 Gate-Jamming and Innate Immune Evasion in MSS Colorectal Cancer

License: CC BY 4.0 Preprint Archive: OSF Dataset: HuggingFace

Anthony J. Vasquez Sr. — Delaware Valley University, Doylestown, PA, USA

Corresponding author: vasquezaj3921@delval.edu


The Paper

Context-Specific Innate Immune Evasion via VDAC1 Gate-Jamming in Microsatellite-Stable Colorectal Cancer: A Three-Cohort Transcriptomic Analysis

Published on Research Square — February 26, 2026. DOI: 10.21203/rs.3.rs-8935902/v1

85-95% of colorectal cancer patients carry microsatellite-stable (MSS) tumors and derive no benefit from immune checkpoint inhibitors. This paper proposes that VDAC1 gate-jamming — suppression of VDAC1 oligomerization by HK-II, Bcl-xL, and mitochondrial cholesterol — silences the cGAS-STING innate immune signal that checkpoint inhibitors require to function, and tests this hypothesis across three cohorts.

Manuscript: paper/gjs_manuscript.docx | paper/gjs_manuscript.md


The Three-Cohort Arc

Analysis Cohort n Result
S1 TCGA pan-cancer 10,071 Null — cross-cancer confounds mask signal
S2 COADREAD MSS/TP53-wt 209 5 Bonferroni-significant immune correlations
S3 IMvigor210 atezolizumab 348 Null — wrong tumor type, wrong treatment mechanism

The two nulls define the framework's boundary. The signal appears precisely where the biology predicts it should — and not where it shouldn't.

S2 Key Findings (MSS/TP53-wildtype clean room)

Marker Spearman rho p_bonf Direction
HAVCR2 (TIM-3) -0.349 5 x 10^-6 Down — T cell absence, not exhaustion
TREX1 +0.315 7 x 10^-5 Up — belt-and-suspenders DNA erasure
CXCL10 -0.231 0.015 Down — suppressed IFN-gamma signaling
STING ratio -0.216 0.034 Down — shift to immunosuppressive profile
cGAS -0.208 0.049 Down

The Hypothesis

tGJS = 0.40 x norm(HK2) + 0.30 x norm(BCL2L1) + 0.30 x norm(TSPO)

A three-gene transcriptomic proxy for the physical Gate-Jamming Score:

GJS = f_HKII x 0.40 + f_BclxL x 0.30 + [Chol]/[CL]_norm x 0.30

Where f_HKII and f_BclxL are the fractions of VDAC1 occupied by hexokinase-II and Bcl-xL respectively (measurable by proximity ligation assay), and [Chol]/[CL] is the outer mitochondrial membrane cholesterol-to-cardiolipin ratio (measurable by lipidomics).

The Therapeutic Stack

Three independent AI analytical systems (Claude Opus, Gemini Pro, Grok) converged on the same three-layer intervention from the same data without cross-exposure:

  1. VDAC1 gate-opener — displace HK-II (methyl jasmonate, clotrimazole) to restore mtDNA release and cGAS-STING activation
  2. DNA/cGAMP eraser inhibitor — block TREX1 or ENPP1 to sustain the innate signal
  3. Checkpoint blockade — amplify the adaptive response now that T cells are being recruited

Repository Structure

vdac-pharmacology-atlas/
├── paper/
│   ├── gjs_manuscript.md                        # Manuscript source
│   ├── gjs_manuscript.docx                      # Submission file
│   ├── build_manuscript_docx.py                 # Reproducible build script
│   ├── supplementary_S1_tcga_boundary_analysis.md
│   ├── supplementary_S2_coadread_mss_stratification.md
│   ├── supplementary_S3_imvigor210.md
│   ├── supplementary-S1-tcga-boundary-analysis.docx
│   ├── supplementary-S2-coadread-mss.docx
│   ├── supplementary-S3-imvigor210.docx
│   └── vpa_cbd_hepatotoxicity_alert.md          # Pharmacovigilance alert
├── analysis/
│   ├── tcga_gjs/
│   │   ├── compute_tgjs.py                      # S1: pan-cancer pipeline
│   │   ├── compute_tgjs_coadread_mss.py         # S2: COADREAD MSS stratified
│   │   ├── data/
│   │   │   └── coadread_mss/                    # S2 results + strata summary
│   │   └── figures/
│   │       ├── fig1-4 (S1 pan-cancer figures)
│   │       └── coadread_mss/ (figS2a-e)
│   └── imvigor210/
│       ├── compute_tgjs_imvigor210.R            # S3: IMvigor210 analysis
│       ├── data/                                 # S3 results matrix + summary
│       └── figures/ (figS3a-d)
├── runs/                                         # IRIS Gate Evo convergence runs
├── gold/                                         # Gold extractions from runs
├── data/
│   ├── vdac_modulators.csv                      # 17 VDAC-interacting compounds
│   └── vdac_isoform_comparison.csv
└── figures/                                      # Atlas overview figures

Supplementary Analyses

File Contents
S1 Pan-cancer tGJS across 10,071 TCGA samples; boundary analysis; ENPP1 anti-correlation
S2 COADREAD 4-stratum analysis; 20 markers x 4 strata; TREX1 co-occurrence finding; ENPP1 artifact correction
S3 IMvigor210 null result; TMB interaction; immune phenotype stratification; OS analysis

Pharmacovigilance Alert

paper/vpa_cbd_hepatotoxicity_alert.md

Up to 30% of co-prescribed VPA + CBD patients develop ALT elevations >3x ULN. The standard CYP450 explanation is incomplete. VPA depletes mitochondrial reserves; CBD alters VDAC1 conductance. An estimated 2,000-3,500 US pediatric patients currently affected. Three testable steps to close the mechanistic gap.


Key Commits

Commit Description
88ff1f7 S1 pan-cancer tGJS pipeline — 10,071 samples, null result
af63c2b S2 COADREAD MSS stratified analysis — 5 Bonferroni hits
af514d1 S2 writeup with synthesis sentence
edb4880 S3 IMvigor210 analysis — second null
61048d4 S3 writeup
779f399 Manuscript restructured around S1-S2-S3 arc
aeb2e44 Submission-ready docx with proper table formatting

Related Projects

Project Link Relationship
VDAC1 Gate-Opening Stack templetwo/VDAC1-Gate-Opening-Therapeutic-Stack Experimental architecture, bench protocols, GJS simulations
IRIS Gate Evo templetwo/iris-gate-evo Multi-model convergence engine
CBD Two-Pathway Model templetwo/cbd-two-pathway-model Origin of the gate-jamming question
IRIS Evo Findings templetwo/iris-evo-findings Run corpus and cross-run analysis
HuggingFace Dataset TheTempleofTwo/vdac-pharmacology-atlas Full convergence corpus
OSF Archive osf.io/c9rqb Preregistration and data archive

Citation

@article{vasquez2026gatejamming,
  title={Context-Specific Innate Immune Evasion via VDAC1 Gate-Jamming
         in Microsatellite-Stable Colorectal Cancer},
  author={Vasquez, Anthony J.},
  year={2026},
  doi={10.21203/rs.3.rs-8935902/v1},
  url={https://doi.org/10.21203/rs.3.rs-8935902/v1},
  journal={Research Square (Preprint)}
}

License

CC BY 4.0. Use it, build on it, cite it.

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An open-access map of the voltage-dependent anion channel as a druggable decision gate

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