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                        v 2.0

Oncogenic Reversion via Attractor-guided Computational Landscape Engineering

Python PyTorch RDKit License Status Models

Can cancer be persuaded to forget it is cancer?


The Hypothesis

Every cancer cell is trapped in a pathological attractor — a stable gene expression state maintained by self-reinforcing epigenetic circuits. Normal tissue identity is a different attractor in the same cell's regulatory landscape, separated by an epigenetic barrier written by enzymes like EZH2 and BRD4.

ORACLE asks: what is the minimal set of transcription factor perturbations that, when applied simultaneously via bifunctional epigenetic molecules, would push a cancer cell over that barrier and lock it into the normal attractor?

This is not killing cancer. It is reverting it.

The approach is grounded in the KAIST REVERT proof-of-concept (Shin et al. 2025), which demonstrated transcriptional identity reversion in colorectal cancer. ORACLE is a computational framework to generalize that result to any cancer type, any patient, and produce drug-like molecule designs (TCIPs) to execute it.


What ORACLE Does

scRNA-seq / scATAC-seq                         Drug-like TCIP molecules
       ↓                                               ↑
[ MODULE 0: Preprocessing ]     →    [ MODULE 2: TCD (Molecule Design) ]
       ↓                                               ↑
[ MODULE 1A: CAM ] → Cancer Attractor              RSP Output
[ MODULE 1B: RSP ] → Reversion Switch Set  ────────────┘
Module Full Name What It Does
CAM Cancer Attractor Mapper Infers the GRN topology and identifies the gene expression attractor state from scRNA-seq. Uses GNN + pseudotime + Boolean ODE simulation.
RSP Reversion Switch Predictor Finds the minimum set of TF activations/repressions that drive the cancer attractor to the normal attractor. Combinatorial search over GRN perturbations.
TCD Transcriptional CIP Designer Designs PROTAC-like bifunctional molecules (TCIPs) that recruit epigenetic writers/erasers to each TF locus. Assembles warhead + linker + recruiter.

Architecture

oracle/
├── cam/                        # Cancer Attractor Mapping
│   ├── cam_pipeline.py         # Orchestrator: scRNA → attractor state
│   ├── grn_inference.py        # GRN reconstruction (GENIE3 / correlation)
│   ├── attractor_finder.py     # Energy landscape + basin identification
│   ├── attractor_classifier.py # ML classifier: cancer vs. normal cell state
│   ├── boolean_network.py      # Boolean ODE network simulation
│   ├── continuous_ode.py       # Continuous ODE for attractor convergence
│   ├── landscape_computer.py   # Waddington potential surface computation
│   ├── pseudotime.py           # Diffusion pseudotime / trajectory inference
│   └── preprocessing.py        # CAM-specific data prep
│
├── rsp/                        # Reversion Switch Prediction
│   ├── rsp_pipeline.py         # Orchestrator: attractor → switch set
│   ├── cancer_score.py         # Differentiability scoring (cancer vs. normal)
│   ├── combinatorial_search.py # Beam search over TF perturbation combos
│   ├── gnn_predictor.py        # GNN: predicts reversion probability
│   ├── perturbation_sim.py     # In-silico perturbation & trajectory sim
│   ├── switch_optimizer.py     # Optimizes the switch set size vs. efficacy
│   ├── druggability_filter.py  # Filters for TF druggability (pocket score)
│   └── trajectory_tracker.py  # Tracks cell trajectory under perturbation
│
├── tcd/                        # TCIP Molecule Design
│   ├── tcd_pipeline.py         # Orchestrator: switch set → TCIP molecules
│   ├── tcip_assembler.py       # 5-tier amide coupling (warhead+linker+recruiter)
│   ├── linker_designer.py      # PEG/alkyl linker library + scoring
│   ├── writer_selector.py      # Writer/eraser selection (p300/BRD4/HDAC2/EZH2/PRMT5)
│   ├── tf_structurer.py        # TF pocket prediction (PDB/AlphaFold)
│   ├── molecule_generator.py   # EGNN-based 3D molecule generation (DDPM)
│   ├── tcip_scorer.py          # Multi-objective molecule scoring
│   ├── ternary_validator.py    # Ternary complex clash/geometry validation
│   └── hard_constraints.py     # Lipinski/Veber/PAINS/Brenk/Ames hard gates
│
├── models/                     # Neural network architectures
│   ├── grn_transformer.py      # Transformer for GRN inference
│   ├── attractor_gnn.py        # GNN: attractor state classification
│   ├── switch_predictor_gnn.py # GNN: switch set prediction
│   ├── cancer_score_mlp.py     # MLP: cancer score function
│   ├── affinity_predictor.py   # MPNN: TF-warhead binding affinity
│   ├── tcip_diffusion.py       # EGNN-DDPM: 3D TCIP generation
│   ├── ternary_complex_predictor.py  # Ternary complex geometry model
│   └── shared/                 # Shared layers (attention, GAT, transformer, SE3)
│
├── preprocessing/              # scRNA-seq preprocessing pipeline
│   ├── scrna_preprocessor.py   # 13-step scRNA pipeline (QC→normalization→HVG)
│   ├── cnv_inference.py        # CNV scoring from expression
│   └── cell_annotator.py       # Cell type annotation
│
├── data/                       # Data loading & fetching
│   ├── fetchers/               # GEO, CellxGene, PDB, AlphaFold, ZINC, TCGA, ENCODE
│   ├── datasets.py             # PyTorch datasets
│   ├── loaders.py              # DataLoader wrappers
│   ├── collators.py            # Batch collation
│   └── samplers.py             # Stratified / balanced samplers
│
├── training/                   # Training infrastructure
│   ├── master_trainer.py       # End-to-end training orchestrator
│   ├── cam_trainer.py          # CAM training loop
│   ├── rsp_trainer.py          # RSP training loop
│   ├── tcd_trainer.py          # TCD training loop
│   ├── losses.py               # All loss functions
│   ├── callbacks.py            # Checkpointing, LR scheduling, early stop
│   └── data_leakage_protocols.py # Patient-level train/test split enforcement
│
├── evaluation/                 # Evaluation & benchmarking
│   ├── benchmarks.py           # End-to-end pipeline benchmarks
│   ├── cam_eval.py             # Attractor accuracy, ARI, NMI
│   ├── rsp_eval.py             # Switch set reversion probability
│   └── tcd_eval.py             # TCIP validity, QED, SA, docking proxy
│
├── visualization/              # Output visualization
│   ├── landscape_viz.py        # 3D Waddington surface
│   ├── trajectory_viz.py       # Cell trajectory plots (UMAP, diffusion map)
│   ├── network_viz.py          # GRN network visualization
│   ├── molecule_viz.py         # TCIP 2D/3D structure rendering
│   └── report_generator.py     # Full HTML/PDF report generation
│
└── interfaces.py               # Frozen dataclass contracts (all module I/O)

scripts/
├── run_inference.py            # Full pipeline: GSE accession → TCIP SMILES
├── run_gbm_pipeline.py         # GBM flagship demo (no training required)
├── run_luad_pipeline.py        # LUAD pipeline (GSE131907)
├── run_aml_pipeline.py         # AML pipeline
├── run_atc_pipeline.py         # Anaplastic thyroid carcinoma pipeline
├── train_all.py                # Train all three modules end-to-end
├── fetch_data.py               # Download & cache GEO/Census datasets
├── preprocess_all.py           # Preprocess all cached datasets
├── plot_gbm_landscape_3d.py    # 3D Waddington landscape visualization
└── plot_gbm_attractor.py       # 2D attractor map with TCIP table

Quick Start

Installation

# Clone
git clone https://github.com/aesenthilvanan-coder/ORACLE-v2.0.git
cd ORACLE-v2.0

# Create environment (conda recommended)
conda create -n oracle python=3.11 -y
conda activate oracle

# Core dependencies
pip install -r requirements.txt

# Biology stack (scanpy, anndata, cellxgene-census)
pip install -r requirements-bio.txt

# Verify installation
make smoke-test

Run the GBM Demo (No Training Required)

The GBM pipeline is fully self-contained and runs off literature-curated biology. It produces all 8 TCIP SMILES with properties and a 3D Waddington landscape in under 60 seconds.

# Design GBM TCIPs
python scripts/run_gbm_pipeline.py

# Render 3D Waddington landscape
python scripts/plot_gbm_landscape_3d.py

# Outputs appear in outputs/
ls outputs/
# gbm_landscape_3d.png   ← 3D epigenetic landscape
# gbm_attractor_map.png  ← attractor gene grid + TCIP table

Full Pipeline from GEO Accession

# Download + preprocess + infer + design — one command
python scripts/run_inference.py \
    --gse GSE131928 \
    --cancer_type GBM \
    --normal_gse GSE67835 \
    --output_dir outputs/gbm_full/

# With a trained model checkpoint
python scripts/run_inference.py \
    --gse GSE131928 \
    --cancer_type GBM \
    --checkpoint checkpoints/stage1a_best.pt \
    --output_dir outputs/gbm_full/

The GBM Demo: End-to-End Example

ORACLE ships with a complete Glioblastoma (GBM) analysis grounded in Neftel et al. 2019 (Cell, GSE131928). GBM was chosen as the flagship cancer because:

  • Median survival: 14.6 months. No second-line standard of care after recurrence.
  • Cannot be fully resected — a reversion approach is uniquely compelling vs. a kill approach.
  • The SOX2/NES stem-cell axis and GFAP/NEUROD1 mature-identity axis are the most well-validated in cancer biology.

Cancer Attractor vs. Normal Attractor

State GBM (MES+NPC stem) Normal Brain
HIGH SOX2, NES, MYC, TWIST1, EZH2, BRD4, CDK4, EGFR, STAT3, VIM, CDH2 GFAP, NEUROD1, RBFOX1, TUBB3, MAP2, S100B
LOW NEUROD1, RBFOX1, GFAP, TUBB3, MAP2, CDKN2A SOX2, NES, MYC, TWIST1, EZH2, BRD4

GBM oscillates between two co-dominant sub-attractors inside the cancer basin:

  MES sub-state ←─────────── GBM Cancer Basin ───────────→ NPC sub-state
  TWIST1↑ VIM↑ CDH2↑ ZEB1↑                               SOX2↑ NES↑ MYC↑ CDK4↑

Both must be targeted simultaneously — hitting only one allows the other to re-seed.

Reversion Switch Set (RSP Output)

ACTIVATE  →  NEUROD1  RBFOX1  GFAP
REPRESS   →  SOX2  MYC  TWIST1  EZH2  BRD4

Why these 8 genes: EZH2 and BRD4 form a self-reinforcing epigenetic loop that keeps the cancer attractor stable:

BRD4 ──reads──→ H3K27ac at SOX2/MYC SE ──drives──→ SOX2/MYC expression
  ↑                                                        ↓
EZH2 expression ←── MYC transcription ←──────────────────┘
  ↓
H3K27me3 at NEUROD1/GFAP ←── EZH2 activity (silences normal identity)

ORACLE attacks this loop at three points: repressing EZH2/BRD4 expression, flipping the super-enhancers from H3K27ac to H3K27me3, and activating NEUROD1/RBFOX1/GFAP to pull the cell into the normal basin.

TCIP Molecules (TCD Output) — 8/8 Pass · All Tier-1 Amide Assembly

Gene Effect Recruiter Ki (nM) MW logP QED SMILES
NEUROD1 ACTIVATE p300 A-485 10 664 5.06 0.116 CC(CNC(=O)CCOCCOCCNC(=O)Cc1cc2ccccc2[nH]1)COC(=O)N[C@H]1CC[C@@H](c2nc3ccccc3s2)CC1
RBFOX1 ACTIVATE p300 A-485 10 662 5.20 0.139 CC(CNC(=O)CCOCCOCCNC(=O)c1ccc2cnccc2c1)COC(=O)N[C@H]1CC[C@@H](c2nc3ccccc3s2)CC1
GFAP ACTIVATE BRD4 JQ1 77 684 2.68 0.168 Cc1nc2c(-c3cc(C(=O)N4CCCC4)nn3C)c(NC(=O)CCOCCOCCNC(=O)c3ccc(S(N)(=O)=O)cc3)ccc2s1
SOX2 REPRESS PRMT5 GSK 3 686 4.24 0.138 O=C(CCOCCOCCNC(=O)c1ccc(-c2nc3ccccc3[nH]2)cc1)Nc1ccc(S(=O)(=O)N2CCC(n3ccnc3)CC2)cc1
MYC REPRESS PRMT5 GSK 3 713 4.39 0.125 O=C(CCOCCOCCNC(=O)c1ccc(Nc2ncnc3ccccc23)cc1)Nc1ccc(S(=O)(=O)N2CCC(n3ccnc3)CC2)cc1
TWIST1 REPRESS HDAC2 entinostat 1.5 642 3.88 0.048 O=C(CCOCCOCCNC(=O)c1ccc2[nH]ccc2c1)Nc1ccc2[nH]c(C(=O)NCc3ccc(NC(=O)NO)cc3)cc2c1
EZH2 REPRESS PRMT5 GSK 3 669 3.24 0.193 O=C(CCOCCOCCNC(=O)c1cnc(NC2CCCCC2)nc1)Nc1ccc(S(=O)(=O)N2CCC(n3ccnc3)CC2)cc1
BRD4 REPRESS EZH2 EPZ-6438 2.5 681 2.07 0.193 O=C(CCOCCOCCNC(=O)Cc1nc2ccccc2s1)NCC(=O)Nc1ccc(C(=O)N2CCC(N3CCOCC3)CC2)cc1

All molecules: MW 640–713 Da · SA ≤ 3.5 · logP 2.1–5.2 · Connected single fragment · bRo5 PROTAC space


Module Deep Dive

Module 1A: CAM

from oracle.cam.cam_pipeline import CAMPipeline

cam = CAMPipeline(config={
    "cancer_type": "GBM",
    "n_hvg": 3000,
    "n_pcs": 50,
    "grn_method": "genie3",
    "n_attractors": 2,
})

cam_output = cam.run(adata, normal_adata=normal_adata)

print(cam_output.cancer_genes_on)   # genes HIGH in cancer attractor
print(cam_output.cancer_genes_off)  # genes LOW in cancer attractor
print(cam_output.attractor_score)   # per-cell cancer attractor score [0, 1]

Internally: 13-step scRNA preprocessing → GRN inference → pseudotime → Boolean ODE simulation → energy landscape → basin identification → normal attractor comparison.

Module 1B: RSP

from oracle.rsp.rsp_pipeline import RSPPipeline

rsp = RSPPipeline(config={
    "max_switch_size": 10,
    "reversion_threshold": 0.7,
    "beam_width": 50,
    "n_simulations": 1000,
})

rsp_output = rsp.run(cam_output)

print(rsp_output.genes_to_activate)          # ['NEUROD1', 'RBFOX1', 'GFAP']
print(rsp_output.genes_to_repress)           # ['SOX2', 'MYC', 'TWIST1', 'EZH2', 'BRD4']
print(rsp_output.validated_reversion_fraction)  # 0.83

Internally: Cancer score gradient → beam search over TF combos → 1000× ODE/Boolean simulation per candidate → reversion fraction scoring → druggability filter → minimal switch set.

Module 2: TCD

from oracle.tcd.tcd_pipeline import TCDPipeline

tcd = TCDPipeline(config={
    "linker_library": "full",
    "max_mw": 1000,
    "require_connected": True,
})

tcd_output = tcd.run(rsp_output)

for tcip in tcd_output.tcip_molecules:
    print(f"{tcip.target_tf:10s} | {tcip.perturbation_type:8s} | "
          f"MW={tcip.molecular_weight:.0f} | QED={tcip.qed:.3f} | "
          f"{'PASS' if tcip.validation_result.passed else 'FAIL'}")
    print(f"  {tcip.full_smiles}")

TCIP assembly architecture:

[TF warhead]─── amide ───[H₂N-Linker-COOH]─── amide ───[Epigenetic recruiter]
     ↑                          ↑                               ↑
Binds TF protein         PEG₂/PEG₃/alkyl             Writer (p300/BRD4/CDK9) or
(HMG/bHLH/SET/RRM)      5–20 heavy atoms             Eraser (HDAC2/EZH2/PRMT5)

5-tier assembly (no fallback for Tiers 1–4):

Tier Strategy
1 warhead-COOH + linker-NH₂ → product-COOH + recruiter-NH₂
2 linker-COOH + warhead-NH₂ → try both second-step orientations
3 Brute-force all 6 permutations of the three fragments
4 Add acetic-acid COOH arm to warhead, then retry Tiers 1–3
5 Force terminal-atom single bond (last resort)

Epigenetic recruiters:

Recruiter Scaffold Ki (nM) Mark Effect
p300 A-485 10 H3K27ac write Activation
BRD4 JQ1 77 H3K27ac amplify Activation
CDK9 AT7519 47 pSer2 RNAPII Activation
MED1 Cortistatin A 300 Super-enhancer Activation
HDAC2 Entinostat 1.5 −H3K27ac Repression
EZH2 EPZ-6438 2.5 H3K27me3 write Repression
PRMT5 GSK3326595 3.0 H4R3me2s Repression
LSD1 Tranylcypromine 243 −H3K4me1 Repression
DNMT3A RG108 115 5mC CpG Repression

Hard Constraints Filter

from oracle.tcd.hard_constraints import TCIPHardConstraints

hc = TCIPHardConstraints()
result = hc.check(smiles, linker_smiles=linker_smiles)

if result.passed:
    print("PASS")
else:
    for v in result.violations:
        print(f"  FAIL: {v}")
Constraint Threshold (bRo5 PROTAC space)
Lipinski MW ≤ 1000 Da
Lipinski logP ≤ 6.0
HBD / HBA ≤ 6 / ≤ 15
Veber RotBonds / TPSA ≤ 25 / ≤ 250 Ų
QED ≥ 0.04
SA Score ≤ 7.0
PAINS / Brenk / Ames 0 alerts

Dataset Guide

Recommended GBM Datasets

GSE Type Description Use
GSE131928 scRNA Neftel 2019 — 28 tumors, canonical MES/NPC/AC/OPC state map Primary GBM attractor
GSE84465 scRNA Darmanis 2017 — first GBM single-cell atlas GRN inference
GSE182109 scRNA Richards 2021 — stem cell hierarchy, 53K cells Sub-basin resolution
GSE162631 scRNA 8 samples, 120K cells (10x) High-density landscape
GSE67835 scRNA Zhang 2016 — normal human brain cell types Normal attractor (required)
GSE163120 scATAC Mack 2022 — GBM chromatin accessibility Enhancer targeting
GSE194329 Spatial GBM Visium — primary + recurrent, IDH-wt Tumor geography
GSE121719 Bulk Primary vs. recurrent paired Attractor shift under SOC

Fetching Data

# Fetch a GEO dataset
python scripts/fetch_data.py --gse GSE131928 --output data/raw/

# Stream from CellxGene Census (no download)
python scripts/fetch_data.py --census --disease "glioblastoma" --n_cells 50000

# Fetch normal brain reference
python scripts/fetch_data.py --gse GSE67835 --output data/raw/normal/

Training

# Stage 0: Molecular pretraining (ZINC22 + PubChem, ~5.6M molecules)
python scripts/run_stage0_only.py --config configs/base_config.yaml

# Stage 1a: Biological pretraining (CellxGene Census, ~2M cancer cells)
python scripts/run_stage1a_census.py --config configs/cam_config.yaml

# Stage 1b + 2: RSP and TCD training
python scripts/train_all.py --checkpoint checkpoints/stage1a_best.pt

Hardware requirements:

Stage Min RAM Runtime (M1 16GB)
Stage 0 (mol pretrain) 8 GB ~12h
Stage 1a (bio pretrain) 16 GB ~24h
Stage 1b (RSP) 8 GB ~4h
Stage 2 (TCD) 8 GB ~3h
Inference only 4 GB < 2 min

Cancer Types Supported

Cancer Script Primary Dataset Key Reversion Targets
GBM run_gbm_pipeline.py GSE131928 SOX2/MYC/TWIST1/EZH2/BRD4 → NEUROD1/RBFOX1/GFAP
LUAD run_luad_pipeline.py GSE131907 MYC/YAP1/ZEB1/EZH2 → FOXA2/NKX2-1
AML run_aml_pipeline.py CellxGene Census MYC/FLT3/EZH2 → CEBPA/PU.1
ATC run_atc_pipeline.py CellxGene Census BRAF effectors/SOX2 → PAX8/FOXE1

To add a new cancer type:

python scripts/run_inference.py \
    --gse YOUR_GSE_ID \
    --cancer_type YOUR_CANCER \
    --normal_gse MATCHED_NORMAL_GSE \
    --output_dir outputs/your_cancer/

Interfaces

All modules communicate through frozen dataclass contracts in oracle/interfaces.py:

from oracle.interfaces import (
    CAMOutput,         # Cancer attractor state + GRN + AnnData
    RSPOutput,         # Switch set (activate/repress) + reversion fraction
    TCDOutput,         # TCIP molecules + validation
    TCIPMolecule,      # Single TCIP: SMILES + properties + validation result
    ValidationResult,  # Hard constraint pass/fail + per-property scores
)

Configuration

# configs/base_config.yaml (excerpt)
model:
  max_params: 514_000_000    # M1 16GB upper bound
  hidden_dim: 512
  n_layers: 8

training:
  batch_size: 64
  learning_rate: 3e-4
  n_epochs: 50
  gradient_clip: 1.0

data:
  n_hvg: 3000
  min_cells: 200

tcd:
  linker_max_mw: 300
  require_connected: true
  max_assembly_tiers: 5      # 1 = strict Tier-1 amide only

Visualizations

# 3D Waddington epigenetic landscape
python scripts/plot_gbm_landscape_3d.py
# → outputs/gbm_landscape_3d.png

# 2D attractor gene grid + TCIP summary table
python scripts/plot_gbm_attractor.py
# → outputs/gbm_attractor_map.png

The 3D landscape renders:

  • GBM cancer basin (left well) with MES and NPC sub-basins
  • Normal brain basin (right well) with astrocyte and neuron sub-basins
  • Epigenetic barrier ridge (EZH2/BRD4 lock)
  • TCIP reversion trajectory (purple) with per-intervention markers

Project Background

ORACLE was built to explore a single scientific question: can we compute a drug-like molecule that forces a cancer cell to re-adopt normal tissue identity?

The key insight from KAIST REVERT (Shin et al. 2025) is that cancer is not only a genetic disease — it is a cellular identity disease. The cancer cell has not lost the code for normal function; it has been pushed into a different attractor in regulatory space and is held there by epigenetic locks. ORACLE's hypothesis is that those locks can be picked computationally.

The TCIP (Transcriptional Cancer Identity Perturbagen) concept is a PROTAC-like bifunctional molecule: one end binds a transcription factor, the other end recruits an epigenetic writer or eraser. Instead of degrading the TF (as PROTACs do), TCIPs rewrite chromatin state at TF-bound loci — converting active enhancers to silenced regions or vice versa, permanently redirecting cell identity.


Citation

@software{oracle2025,
  author  = {Senthilvanan, Aravind E.},
  title   = {{ORACLE}: Oncogenic Reversion via Attractor-guided
             Computational Landscape Engineering},
  year    = 2025,
  version = {2.0},
  url     = {https://github.com/aesenthilvanan-coder/ORACLE-v2.0},
}

Key references:

  • Shin et al. 2025 — REVERT: transcriptional identity reversion in colorectal cancer (KAIST)
  • Neftel et al. 2019 — Integrative model of cellular states in GBM (Cell)
  • Darmanis et al. 2017 — Single-cell characterization of GBM (Nature Neuroscience)
  • Zhang et al. 2016 — Purification of progenitor and mature cells from human brain (Neuron)
  • Filippakopoulos et al. 2010 — Selective inhibition of BET bromodomains (Nature)
  • Konze et al. 2013 — An orally bioavailable chemical probe of EZH2 (ACS Chemical Biology)

License

MIT License — see LICENSE.


"The cancer cell already knows how to be normal. We just need to remind it."

Built with Python · PyTorch · RDKit · scanpy · CellxGene Census

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End-to-end computational framework for designing cancer identity reversion therapies: scRNA-seq → Waddington attractor mapping → epigenetic switch set prediction → TCIP bifunctional molecule design

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