What question did this study set out to answer?

The aim is to test the hypothesis that H3K27me3 enrichment varies at neural crest migration loci between domestic Sus scrofa and wild boar embryos.

March 21, 2026Open Access

Pre-Registration P9: H3K27me3 Enrichment at Neural Crest Migration Loci in Domestic Sus scrofa vs. Wild Boar Embryos — The ChIP-seq Experiment

Key Points

The aim is to test the hypothesis that H3K27me3 enrichment varies at neural crest migration loci between domestic Sus scrofa and wild boar embryos.
Conduct a ChIP-seq experiment on E14-E18 domestic Sus scrofa and wild boar embryos.
Measure H3K27me3 enrichment at nine pre-registered NCC migration driver loci.
Use a minimum enrichment ratio criterion and include negative control loci.
At least 4 of 9 target loci are expected to show significant enrichment in domestic compared to wild boar embryos.
Negative controls (GAPDH, ACTB, MYF5, HBB) should not show differential enrichment.
RNA-seq validation aims for functional silencing consistency in at least 3 confirmed loci.

Abstract

This document is a pre-registered experimental protocol deposited before any sequencing data has been generated. It is locked at the time of Zenodo deposit and may not be modified after that point. The field-arrest model of domestication (Lawson, 2026, doi:10.5281/zenodo.19094935) proposes that the domestication syndrome is produced by EZH2-mediated H3K27me3 deposition at neural crest cell (NCC) migration driver loci during embryogenesis, arresting NCC migration at the juvenile-proximate developmental stage. This is the same molecular mechanism that maintains cancer attractor arrest in triple-negative breast cancer (TNBC), where EZH2 deposits H3K27me3 at FOXA1, GATA3, and ESR1, silencing the luminal identity program. This pre-registration specifies the ChIP-seq experiment that tests the molecular mechanism prediction. Domestic Sus scrofa embryos at gestational day E14--E18 (peak NCC active migration window) are predicted to show significantly elevated H3K27me3 enrichment at the following pre-registered NCC migration driver loci relative to wild boar embryos at the same developmental stage: BMP4, BMP7, WNT1, WNT3A, CXCL12, CXCR4, EDNRB, SOX10, SNAI2. Primary success criterion: minimum 4 of 9 target loci reach enrichment ratio ≥ 1.5 (domestic/wild boar) at q < 0.05 (FDR-corrected), with negative control loci (GAPDH, ACTB, MYF5, HBB) showing no differential enrichment, and RNA-seq concordance confirming functional silencing at ≥ 3 confirmed loci. The genetic model predicts no differential H3K27me3 at NCC loci between domestic and wild boar embryos at equivalent developmental stages. The field-arrest model predicts specific locus-directed enrichment. The models are distinguished by this experiment. P9 is the gateway experiment for P11 (tazemetostat in pregnant domestic sow, shifting F1 morphology toward wild-type). P11 is not justified until P9 confirms EZH2 as the arrest maintenance enzyme. If P9 confirms, the same drug (tazemetostat) that de-arrests cancer cells by removing H3K27me3 at identity loci is predicted to de-arrest domestic pig embryos by removing H3K27me3 at NCC migration loci. Same enzyme. Same mark. Same drug. Different scale. Pre-specified failure diagnoses (F1--F4) are included. A null result is informative and will be published in full. Theory paper: doi:10.5281/zenodo.19094935 P7 pre-registration: doi:10.5281/zenodo.19096128 Repository: github.com/Eric-Robert-Lawson/attractor-oncology

Pre-Registration P9: H3K27me3 Enrichment at Neural Crest Migration Loci in Domestic Sus scrofa vs. Wild Boar Embryos — The ChIP-seq Experiment

Key Points

Abstract

Cite This Study