Extract context-specific region embeddings from ChromBERT

This notebook shows how to extract region embeddings using the ChromBERT-tools Python API.

embed_region API: Generate region embeddings for input regions.

umap_plot API: Visualize region embeddings with UMAP using user-defined annotations.

For the bash command-line usage, see `examples/cli/embed_region.ipynb <../cli/embed_region.ipynb>`__

For more details, please refer to the `embed_region <https://chrombert-tools.readthedocs.io/en/latest/commands/embed_region.html>`__ command documentation

[1]:
from chrombert_tools import embed_region, umap_plot

generate region embedding

[2]:
import numpy as np
import pandas as pd
[3]:
# Returns:
# - region_emb: Region embedding array with shape ``(n_regions, 768)``. Each row corresponds to one ChromBERT bin overlapping the input regions.
# - overlap_region_bed: Input regions that overlap ChromBERT bins.
#   build_region_index maps to ChromBERT's reference regions

results = embed_region(
    region="../data/umap_region_1kb_downsample.bed", # your region
    odir="./output_emb_region_1kb", # output directory
    genome="hg38",        # Options: "hg38", "mm10"
    resolution="1kb",     # Options: "1kb", "2kb", "4kb", "200bp"
)
region_emb=results.region_emb
overlap_region_bed=results.overlap_region_bed
Region summary - total: 3000, overlapping with ChromBERT: 3000 (one region may overlap multiple ChromBERT regions, we keep overlaps with ≥50% coverage of either the ChromBERT bin or the input region), non-overlapping: 0
Using cached region embeddings...

Finished!
Focus region summary - total: 3000, overlapping with ChromBERT: 3000, non-overlapping: 0
Note: It is possible for a single region to overlap multiple ChromBERT regions.
Overlapping regions BED file: ./output_emb_region_1kb/overlap_region.bed
Non-overlapping regions BED file: ./output_emb_region_1kb/no_overlap_region.bed
Region embeddings saved to: ./output_emb_region_1kb/region_emb_emb.npy
Embedding type: general
[4]:
# Region embedding array with shape ``(n_regions, 768)``.
# Each row corresponds to one ChromBERT bin overlapping the input regions.
region_emb
[4]:
array([[ 3.547 ,  0.828 ,  1.625 , ..., -0.578 , -1.422 , -1.406 ],
       [-0.7812, -0.4355,  0.621 , ..., -0.59  , -0.8867,  0.6367],
       [-0.9375, -0.2031,  1.016 , ..., -0.377 , -0.2637,  1.547 ],
       ...,
       [-0.336 , -0.9375,  1.07  , ..., -1.234 , -0.953 , -0.5117],
       [ 0.0398,  0.287 , -0.2012, ...,  0.2812,  0.0908,  1.781 ],
       [ 0.1543, -0.4746,  1.281 , ..., -0.711 , -0.668 , -0.252 ]],
      dtype=float16)
[5]:
# Input regions that overlap ChromBERT bins.
# DataFrame columns: [chrom, start, end, build_region_index, start_input, end_input].
# The region order matches the order of embeddings in `region_emb`.
overlap_region_bed
[5]:
chrom start end build_region_index start_input end_input
0 chr1 54000 55000 6 54000 55000
1 chr1 916000 917000 213 916000 917000
2 chr1 960000 961000 255 960000 961000
3 chr1 1004000 1005000 296 1004000 1005000
4 chr1 1013000 1014000 305 1013000 1014000
... ... ... ... ... ... ...
2995 chr1 248839000 248840000 183893 248839000 248840000
2996 chr1 248857000 248858000 183907 248857000 248858000
2997 chr1 248861000 248862000 183911 248861000 248862000
2998 chr1 248873000 248874000 183923 248873000 248874000
2999 chr1 248945000 248946000 183981 248945000 248946000

3000 rows × 6 columns

[6]:
# Map each region to its user-defined annotation class

region_anno = pd.read_csv("../data/umap_region_1kb_downsample.bed",sep="\t",header=None,names=["chrom","start_input","end_input","build_region_index","anno"])
overlap_region_bed = overlap_region_bed.merge(region_anno[["chrom","start_input","end_input","anno"]],on=["chrom","start_input","end_input"])
overlap_region_bed

[6]:
chrom start end build_region_index start_input end_input anno
0 chr1 54000 55000 6 54000 55000 repressive
1 chr1 916000 917000 213 916000 917000 promoter
2 chr1 960000 961000 255 960000 961000 promoter
3 chr1 1004000 1005000 296 1004000 1005000 promoter
4 chr1 1013000 1014000 305 1013000 1014000 promoter
... ... ... ... ... ... ... ...
2995 chr1 248839000 248840000 183893 248839000 248840000 promoter
2996 chr1 248857000 248858000 183907 248857000 248858000 promoter
2997 chr1 248861000 248862000 183911 248861000 248862000 repressive
2998 chr1 248873000 248874000 183923 248873000 248874000 promoter
2999 chr1 248945000 248946000 183981 248945000 248946000 repressive

3000 rows × 7 columns

[7]:
# umap plot
umap_plot(region_emb, overlap_region_bed["anno"], "./output_emb_region_1kb")
/mnt/Storage/home/chenqianqian/miniconda3/envs/chrombert/lib/python3.9/site-packages/umap/umap_.py:1952: UserWarning: n_jobs value 1 overridden to 1 by setting random_state. Use no seed for parallelism.
  warn(
../../_images/examples_api_embed_region_8_1.png

generate gene (promoter region) embeddings

[8]:
# gene_emb_dict: Python dictionary mapping each matched gene to a 768-dimensional embedding.
results = embed_region(
    gene="ENSG00000170921;TANC2;ENSG00000200997;DPYD;SNORA70;tp53;brd4", # your genes
    odir="./output_emb_genes", # output directory
    genome="hg38",        # Options: "hg38", "mm10"
    resolution="1kb",     # Options: "1kb", "2kb", "4kb", "200bp"
)
gene_emb_dict = results.gene_emb_dict
Using cached region embeddings for gene pooling...

Finished!
Note: All gene names were converted to lowercase for matching.
Gene count summary - requested: 7, matched: 7, not found: 0
Matched gene meta saved to: ./output_emb_genes/overlap_genes_meta.tsv
Gene embeddings saved to: ./output_emb_genes/gene_emb_emb.pkl
Embedding type: general
[9]:
# Python dictionary mapping each matched gene to a 768-dimensional embedding.
for key, value in gene_emb_dict.items():
    print(key,value.shape)
ensg00000170921 (768,)
tanc2 (768,)
ensg00000200997 (768,)
dpyd (768,)
snora70 (768,)
tp53 (768,)
brd4 (768,)

Generate cell-type-specific embeddings

We use cell-type-specific chromatin accessibility peak and signal files as input.

The embed_region API uses these data to build a cell-type-specific model and generate cell-type-specific embeddings.

[10]:
import os
os.environ["CUDA_VISIBLE_DEVICES"]='0'
[ ]:
# download example data
# Myoblast cell-type-specific chromatin accessibility peak and signal files


import subprocess
if not os.path.exists('../data/myoblast_ENCFF647RNC_peak.bed'):
    cmd = f'wget https://www.encodeproject.org/files/ENCFF647RNC/@@download/ENCFF647RNC.bed.gz -O ../data/myoblast_ENCFF647RNC_peak.bed.gz'
    subprocess.run(cmd, shell=True)
    cmd = f"gzip -d ../data/myoblast_ENCFF647RNC_peak.bed.gz"
    subprocess.run(cmd, shell=True)

# import subprocess
if not os.path.exists('../data/myoblast_ENCFF149ERN_signal.bigwig'):
    cmd = f'wget https://www.encodeproject.org/files/ENCFF149ERN/@@download/ENCFF149ERN.bigWig -O ../data/myoblast_ENCFF149ERN_signal.bigwig'
    subprocess.run(cmd, shell=True)
[ ]:
# We only use the first 100 lines of the peak file for demonstration
!head -n 100 ../data/myoblast_ENCFF647RNC_peak.bed > ../data/myoblast_ENCFF647RNC_peak_100.bed
[13]:
# Generate cell-type-specific embeddings using chromatin accessibility peak and signal files as input
# Build a cell-type-specific model from the input data
# Generate embeddings using the fine-tuned model
results_myoblast_specific = embed_region(
    region = "../data/myoblast_ENCFF647RNC_peak_100.bed", # your focus region
    odir = "./output_cell_specific_emb_train", # output directory
    cell_type_bw = "../data/myoblast_ENCFF149ERN_signal.bigwig", # your focus cell-type accessibility data
    cell_type_peak = "../data/myoblast_ENCFF647RNC_peak.bed", # your focus cell-type peak data
    genome = "hg38", # genome
    resolution = "1kb", # resolution
)

Preparing dataset ...
Region summary - total: 373422, overlapping with ChromBERT: 368260 (one region may overlap multiple ChromBERT regions, we keep overlaps with ≥50% coverage of either the ChromBERT bin or the input region), non-overlapping: 7920
Total regions: 324690
Fast mode: downsampling to 20k regions
Fine-tuning cell-specific model...

[Attempt 0/2] seed=55
Load pretrained ckpt /mnt/Storage/home/chenqianqian/.cache/chrombert/data/checkpoint/hg38_6k_1kb_pretrain.ckpt successfully!
/mnt/Storage/home/chenqianqian/miniconda3/envs/chrombert/lib/python3.9/site-packages/torchmetrics/utilities/prints.py:43: UserWarning: Metric `SpearmanCorrcoef` will save all targets and predictions in the buffer. For large datasets, this may lead to large memory footprint.
  warnings.warn(*args, **kwargs)
Using bfloat16 Automatic Mixed Precision (AMP)
GPU available: True (cuda), used: True
TPU available: False, using: 0 TPU cores
HPU available: False, using: 0 HPUs
You are using a CUDA device ('NVIDIA A100-PCIE-40GB') that has Tensor Cores. To properly utilize them, you should set `torch.set_float32_matmul_precision('medium' | 'high')` which will trade-off precision for performance. For more details, read https://pytorch.org/docs/stable/generated/torch.set_float32_matmul_precision.html#torch.set_float32_matmul_precision
LOCAL_RANK: 0 - CUDA_VISIBLE_DEVICES: [0]
Loading `train_dataloader` to estimate number of stepping batches.
/mnt/Storage/home/chenqianqian/miniconda3/envs/chrombert/lib/python3.9/site-packages/lightning/pytorch/utilities/model_summary/model_summary.py:231: Precision bf16-mixed is not supported by the model summary.  Estimated model size in MB will not be accurate. Using 32 bits instead.

  | Name  | Type             | Params | Mode
---------------------------------------------------
0 | model | ChromBERTGeneral | 62.8 M | train
---------------------------------------------------
18.9 M    Trainable params
43.9 M    Non-trainable params
62.8 M    Total params
251.095   Total estimated model params size (MB)
153       Modules in train mode
0         Modules in eval mode
/mnt/Storage/home/chenqianqian/miniconda3/envs/chrombert/lib/python3.9/site-packages/lightning/pytorch/trainer/connectors/data_connector.py:484: Your `val_dataloader`'s sampler has shuffling enabled, it is strongly recommended that you turn shuffling off for val/test dataloaders.
Metric default_validation/pcc improved. New best score: 0.269
Metric default_validation/pcc improved by 0.166 >= min_delta = 0.01. New best score: 0.435
Metric default_validation/pcc improved by 0.089 >= min_delta = 0.01. New best score: 0.524
Metric default_validation/pcc improved by 0.085 >= min_delta = 0.01. New best score: 0.609
Metric default_validation/pcc improved by 0.133 >= min_delta = 0.01. New best score: 0.742
Metric default_validation/pcc improved by 0.022 >= min_delta = 0.01. New best score: 0.764
Metric default_validation/pcc improved by 0.068 >= min_delta = 0.01. New best score: 0.832
Metric default_validation/pcc improved by 0.010 >= min_delta = 0.01. New best score: 0.842
Metric default_validation/pcc improved by 0.018 >= min_delta = 0.01. New best score: 0.860
Metric default_validation/pcc improved by 0.012 >= min_delta = 0.01. New best score: 0.872
Metric default_validation/pcc improved by 0.013 >= min_delta = 0.01. New best score: 0.885
Metric default_validation/pcc improved by 0.017 >= min_delta = 0.01. New best score: 0.902
Monitored metric default_validation/pcc did not improve in the last 5 records. Best score: 0.902. Signaling Trainer to stop.
`Trainer.fit` stopped: `max_epochs=5` reached.
Evaluating the finetuned model performance
Load pretrained ckpt /mnt/Storage/home/chenqianqian/.cache/chrombert/data/checkpoint/hg38_6k_1kb_pretrain.ckpt successfully!
Loading checkpoint from /mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt
Loading from pl module, remove prefix 'model.'
Loading from pl module, replace 'pretrain_model' with 'pretrain_model.chrombert'
Loaded 111/111 parameters
/mnt/Storage/home/chenqianqian/miniconda3/envs/chrombert/lib/python3.9/site-packages/torchmetrics/utilities/prints.py:43: UserWarning: Metric `SpearmanCorrcoef` will save all targets and predictions in the buffer. For large datasets, this may lead to large memory footprint.
  warnings.warn(*args, **kwargs)
ft_ckpt: /mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt, test_metrics: {'pearsonr': 0.8920528888702393, 'spearmanr': 0.8251720666885376, 'mse': 0.028753146529197693, 'mae': 0.0986708402633667, 'r2': 0.7648238203557352}
Attempt metrics: pearsonr=0.8920528888702393
Accepted run (pearsonr=0.8921 >= 0.4).

Finished stage 2: obtained a fine-tuned ChromBERT
Best pearsonr=0.8920528888702393, metrics={'pearsonr': 0.8920528888702393, 'spearmanr': 0.8251720666885376, 'mse': 0.028753146529197693, 'mae': 0.0986708402633667, 'r2': 0.7648238203557352, 'ft_ckpt': '/mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt'}
Region summary - total: 100, overlapping with ChromBERT: 101 (one region may overlap multiple ChromBERT regions, we keep overlaps with ≥50% coverage of either the ChromBERT bin or the input region), non-overlapping: 0
Your supervised_file does not contain the 'label' column. Please verify whether ground truth column ('label') is required. If it is not needed, you may disregard this message.
100%|██████████| 26/26 [00:02<00:00, 12.59it/s]

Finished!
Focus region summary - total: 100, overlapping with ChromBERT: 101, non-overlapping: 0
Note: It is possible for a single region to overlap multiple ChromBERT regions.
Overlapping regions BED file: ./output_cell_specific_emb_train/overlap_region.bed
Non-overlapping regions BED file: ./output_cell_specific_emb_train/no_overlap_region.bed
Region embeddings saved to: ./output_cell_specific_emb_train/region_emb_emb.npy
Embedding type: cell-specific

[14]:
region_emb = results_myoblast_specific.region_emb # cell-type-specific region embeddings
region_emb
[14]:
array([[ 1.2109375 , -0.06298828,  0.66015625, ..., -0.19921875,
        -1.0234375 , -0.453125  ],
       [-0.19433594, -0.02001953,  1.21875   , ..., -0.38476562,
        -0.5625    ,  0.45507812],
       [ 2.171875  ,  0.3515625 ,  0.38867188, ..., -0.41796875,
        -1.390625  , -0.68359375],
       ...,
       [-1.265625  , -0.6328125 ,  0.59375   , ..., -0.58203125,
        -1.2109375 ,  0.33984375],
       [-1.5546875 , -0.68359375,  0.58984375, ..., -0.35546875,
        -1.        ,  0.6328125 ],
       [-1.1875    , -0.58203125,  0.66015625, ..., -0.45898438,
        -0.6875    ,  0.6328125 ]], dtype=float32)

Generate cell-type-specific embeddings using a fine-tuned checkpoint

We use a fine-tuned checkpoint as input.

The embed_region API uses this checkpoint to generate cell-type-specific embeddings.

[ ]:
# import glob
# ft_ckpt_dir = "./output_cell_specific_emb_train/train/**/*.ckpt"

# ft_ckpt = glob.glob(ft_ckpt_dir, recursive=True)[0]
ft_ckpt = results_myoblast_specific.model_ckpt  # Path to the cell-type-specific model checkpoint generated in the previous step
[16]:
from chrombert_tools import embed_region
[17]:
# Generate cell-type-specific embeddings using a fine-tuned checkpoint as input
# Loading fine-tuned model using the checkpoint
# Generate embeddings using the fine-tuned model

results_myoblast_specific_2 = embed_region(
    region = "../data/myoblast_ENCFF647RNC_peak_100.bed", # your focus region
    gene="ENSG00000170921;TANC2;ENSG00000200997;DPYD;SNORA70;tp53;brd4", # your genes
    odir = "./output_cell_specific_emb_cistrome_2", # output directory
    ft_ckpt = ft_ckpt, # fine-tuned checkpoint
    genome = "hg38", # genome
    resolution = "1kb", # resolution
)

Using provided fine-tuned checkpoint: /mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt
Load pretrained ckpt /mnt/Storage/home/chenqianqian/.cache/chrombert/data/checkpoint/hg38_6k_1kb_pretrain.ckpt successfully!
Loading checkpoint from /mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt
Loading from pl module, remove prefix 'model.'
Loading from pl module, replace 'pretrain_model' with 'pretrain_model.chrombert'
Loaded 111/111 parameters
Region summary - total: 100, overlapping with ChromBERT: 101 (one region may overlap multiple ChromBERT regions, we keep overlaps with ≥50% coverage of either the ChromBERT bin or the input region), non-overlapping: 0
Your supervised_file does not contain the 'label' column. Please verify whether ground truth column ('label') is required. If it is not needed, you may disregard this message.
100%|██████████| 26/26 [00:02<00:00, 12.52it/s]

Finished!
Focus region summary - total: 100, overlapping with ChromBERT: 101, non-overlapping: 0
Note: It is possible for a single region to overlap multiple ChromBERT regions.
Overlapping regions BED file: ./output_cell_specific_emb_cistrome_2/overlap_region.bed
Non-overlapping regions BED file: ./output_cell_specific_emb_cistrome_2/no_overlap_region.bed
Region embeddings saved to: ./output_cell_specific_emb_cistrome_2/region_emb_emb.npy
Embedding type: cell-specific
Your supervised_file does not contain the 'label' column. Please verify whether ground truth column ('label') is required. If it is not needed, you may disregard this message.
100%|██████████| 7/7 [00:00<00:00,  8.87it/s]

Finished!
Note: All gene names were converted to lowercase for matching.
Gene count summary - requested: 7, matched: 7, not found: 0
Matched gene meta saved to: ./output_cell_specific_emb_cistrome_2/overlap_genes_meta.tsv
Gene embeddings saved to: ./output_cell_specific_emb_cistrome_2/gene_emb_emb.pkl
Embedding type: cell-specific

[18]:
results_myoblast_specific_2.region_emb
[18]:
array([[ 1.2109375 , -0.06298828,  0.66015625, ..., -0.19921875,
        -1.0234375 , -0.453125  ],
       [-0.19433594, -0.02001953,  1.21875   , ..., -0.38476562,
        -0.5625    ,  0.45507812],
       [ 2.171875  ,  0.3515625 ,  0.38867188, ..., -0.41796875,
        -1.390625  , -0.68359375],
       ...,
       [-1.265625  , -0.6328125 ,  0.59375   , ..., -0.58203125,
        -1.2109375 ,  0.33984375],
       [-1.5546875 , -0.68359375,  0.58984375, ..., -0.35546875,
        -1.        ,  0.6328125 ],
       [-1.1875    , -0.58203125,  0.66015625, ..., -0.45898438,
        -0.6875    ,  0.6328125 ]], dtype=float32)
[19]:
results_myoblast_specific_2.model_ckpt
[19]:
'/mnt/Storage2/home/chenqianqian/projects/chrombert/chrombert_tools/ChromBERT-tools/examples/api/output_cell_specific_emb_train/train/try_00_seed_55/lightning_logs/lightning_logs/version_0/checkpoints/epoch=3-step=239.ckpt'
[20]:
results_myoblast_specific_2.gene_emb_dict
[20]:
{'ensg00000170921': array([-7.18750000e-01, -3.10058594e-02,  7.07031250e-01, -1.67236328e-02,
        -9.88281250e-01, -3.06640625e-01,  8.16406250e-01,  3.82812500e-01,
         1.01562500e+00,  5.74218750e-01, -4.95605469e-02,  6.21093750e-01,
        -6.40625000e-01, -2.98828125e-01,  6.32812500e-01,  3.14941406e-02,
         1.46875000e+00,  7.81250000e-01, -5.31250000e-01, -9.92187500e-01,
         6.79687500e-01,  5.42968750e-01, -9.10156250e-01,  1.36718750e-01,
        -4.31640625e-01,  8.90625000e-01, -4.35546875e-01,  7.38281250e-01,
         1.73828125e-01, -3.84765625e-01, -2.98828125e-01, -8.51562500e-01,
         9.02343750e-01, -2.57812500e-01, -5.29785156e-02,  6.52343750e-01,
         1.00000000e+00,  1.40625000e+00,  3.94531250e-01, -5.22460938e-02,
         1.11718750e+00,  4.55078125e-01, -9.13085938e-02,  2.89062500e-01,
        -3.78906250e-01, -9.47265625e-02, -1.23291016e-02,  1.42578125e-01,
         1.85546875e-01, -1.26953125e-01, -1.61132812e-01, -2.81250000e-01,
         3.06640625e-01, -5.62500000e-01,  3.49121094e-02,  2.32421875e-01,
         8.32031250e-01,  2.38281250e-01,  3.45703125e-01,  3.73046875e-01,
        -2.33398438e-01,  2.65625000e-01,  8.32031250e-01,  3.33984375e-01,
        -2.87109375e-01, -6.01562500e-01, -4.88281250e-01,  1.01562500e+00,
         5.82031250e-01, -8.49609375e-02,  3.22265625e-01,  3.37890625e-01,
        -3.51562500e-01,  4.19921875e-01, -4.12109375e-01,  3.60107422e-03,
         4.57031250e-01,  5.35156250e-01, -8.78906250e-01,  8.39843750e-01,
         2.65625000e-01,  9.80468750e-01,  6.40625000e-01, -5.54687500e-01,
         1.92187500e+00,  8.98437500e-02, -3.75000000e-01,  5.07812500e-01,
         2.50000000e-01,  9.42382812e-02, -2.79541016e-02, -6.34765625e-02,
        -8.75000000e-01, -2.46093750e-01, -1.96533203e-02,  9.25781250e-01,
        -7.38281250e-01, -1.12500000e+00,  1.02343750e+00,  1.84570312e-01,
        -2.55859375e-01, -3.33984375e-01,  1.16406250e+00,  2.00195312e-01,
        -5.35156250e-01,  1.84570312e-01, -6.13281250e-01, -9.92187500e-01,
         1.19531250e+00,  2.69531250e-01, -2.24609375e-02,  7.42187500e-01,
        -4.43359375e-01, -1.17968750e+00,  1.65625000e+00,  1.37695312e-01,
        -1.94531250e+00,  1.12304688e-01,  4.31640625e-01,  3.51562500e-01,
         1.26562500e+00,  1.70898438e-01, -5.89843750e-01, -7.76367188e-02,
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[ ]:

[ ]: