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{
 "cells": [
  {
   "cell_type": "markdown",
   "id": "70438228",
   "metadata": {},
   "source": [
    "# Data Preprocessing (Interactive Notebook)\n",
    "\n",
    "This notebook runs the repository's Step1→Step9 data-preprocessing steps interactively so you can execute, inspect and debug individual stages locally.\n",
    "\n",
    "Outputs from these steps will be written under `example/data_preprocessing` and downstream under `example/machine_learning` by default."
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "2348a749",
   "metadata": {},
   "outputs": [],
   "source": [
    "%%bash\n",
    "git clone https://github.com/hlnicholls/GenePrioritiser\n",
    "cd GenePrioritiser\n",
    "\n",
    "conda env create -f GenePrioritiser_env.yml\n",
    "conda activate GenePrioritiser_env\n",
    "pip install --force-reinstall scikit-learn==1.4.2\n",
    "pip install --force-reinstall scipy==1.11.4\n",
    "pip install --force-reinstall numpy==1.23.0\n",
    "pip install scikit-optimize\n",
    "conda install -c bioconda bedtools htslib\n",
    "conda install -c conda-forge parallel\n",
    "pip install pybedtools intervaltree requests"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 1,
   "id": "74317926",
   "metadata": {},
   "outputs": [],
   "source": [
    "import os\n",
    "os.chdir(\"/Users/hannahnicholls/GitHub/GenePrioritiser\")"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 2,
   "id": "d9113571",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Using cached gene bed: /Users/hannahnicholls/GitHub/GenePrioritiser/utils/gencode_v19.genes.ext10000.sorted.bed\n",
      "Processing /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_DBP.txt.gz in /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_DBP.txt_27680\n",
      "Parsing GWAS input and extracting SNP positions (Python)...\n",
      "Extracted 7160657 SNP positions from /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_DBP.txt.gz\n",
      "Running bedtools closest (this is the fast step, but can be I/O heavy on large GWAS)...\n",
      "Building gene map (line -> gene, distance)...\n",
      "Joining original rows with gene map and writing outputs via Python...\n",
      "Wrote annotated 7160657 SNPs to ./example/data_preprocessing/output/variants/Annotated_GWAS_DBP.csv\n",
      "Wrote variant data to ./example/data_preprocessing/output/variants/variant_data_DBP.csv\n",
      "Cleaning up temporary files: /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_DBP.txt_27680\n",
      "Done for /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_DBP.txt.gz\n",
      "Processing /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_SBP.txt.gz in /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_SBP.txt_27680\n",
      "Parsing GWAS input and extracting SNP positions (Python)...\n",
      "Extracted 7088121 SNP positions from /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_SBP.txt.gz\n",
      "Running bedtools closest (this is the fast step, but can be I/O heavy on large GWAS)...\n",
      "Building gene map (line -> gene, distance)...\n",
      "Joining original rows with gene map and writing outputs via Python...\n",
      "Wrote annotated 7088121 SNPs to ./example/data_preprocessing/output/variants/Annotated_GWAS_SBP.csv\n",
      "Wrote variant data to ./example/data_preprocessing/output/variants/variant_data_SBP.csv\n",
      "Cleaning up temporary files: /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_SBP.txt_27680\n",
      "Done for /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_SBP.txt.gz\n",
      "Processing /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_PP.txt.gz in /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_PP.txt_27680\n",
      "Parsing GWAS input and extracting SNP positions (Python)...\n",
      "Extracted 7088842 SNP positions from /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_PP.txt.gz\n",
      "Running bedtools closest (this is the fast step, but can be I/O heavy on large GWAS)...\n",
      "Building gene map (line -> gene, distance)...\n",
      "Joining original rows with gene map and writing outputs via Python...\n",
      "Wrote annotated 7088842 SNPs to ./example/data_preprocessing/output/variants/Annotated_GWAS_PP.csv\n",
      "Wrote variant data to ./example/data_preprocessing/output/variants/variant_data_PP.csv\n",
      "Cleaning up temporary files: /Users/hannahnicholls/GitHub/GenePrioritiser/tmp_annotation_Evangelou_30224653_PP.txt_27680\n",
      "Done for /Users/hannahnicholls/GitHub/GenePrioritiser/./example/data_preprocessing/input/Evangelou_30224653_PP.txt.gz\n"
     ]
    }
   ],
   "source": [
    "!bash /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step1_annotate_genes_bedtools.sh"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 3,
   "id": "9ff8bfe6",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Repo root         : /Users/hannahnicholls/GitHub/GenePrioritiser\n",
      "Variant directory : example/data_preprocessing/output/variants\n",
      "HGNC file         : /Users/hannahnicholls/GitHub/GenePrioritiser/utils/hgnc_complete_set.txt\n",
      "Building HGNC synonym map...\n",
      "Loaded 102319 synonym mappings\n",
      "\n",
      "=== Harmonising Annotated_GWAS_* files ===\n",
      "  Processing Annotated_GWAS_DBP.csv ... [OK, 428054 gene names changed]\n",
      "  Processing Annotated_GWAS_PP.csv ... [OK, 423689 gene names changed]\n",
      "  Processing Annotated_GWAS_SBP.csv ... [OK, 423690 gene names changed]\n",
      "\n",
      "=== Harmonising variant_data_* files ===\n",
      "  Processing variant_data_DBP.csv ... [OK, 428054 gene names changed]\n",
      "  Processing variant_data_PP.csv ... [OK, 423689 gene names changed]\n",
      "  Processing variant_data_SBP.csv ... [OK, 423690 gene names changed]\n",
      "\n",
      "Done. All applicable files overwritten with HGNC-harmonised Gene symbols.\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step2_harmonise_genes.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 4,
   "id": "8a20f2dd",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "All files processed and merged. Output saved to: ./databases/variant_level/merged_gene_median_variant_measures.csv\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step3_process_variant_level_data.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 5,
   "id": "1ef583bf",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Number of unmapped entries: 0\n",
      "Sample unmapped entries:\n",
      "Empty DataFrame\n",
      "Columns: [protein1, protein2, coexpression, experiments, database, Gene1, Gene2]\n",
      "Index: []\n",
      "Interactors collected with max_depth=3: 19430 genes\n",
      "Direct only interactors (depth=1): 17554\n",
      "One-level interactors (depth=2): 19428\n",
      "Found 3 annotated GWAS file(s) to load:\n",
      " - ./example/data_preprocessing/output/variants/Annotated_GWAS_DBP.csv\n",
      " - ./example/data_preprocessing/output/variants/Annotated_GWAS_PP.csv\n",
      " - ./example/data_preprocessing/output/variants/Annotated_GWAS_SBP.csv\n",
      "Wrote intermediate least-likely genes to /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_intermediate.tsv (will not overwrite ./example/data_preprocessing/input/least_likely_genes.tsv)\n",
      "Applying additional filtering based on: ./example/data_preprocessing/input/BP_loci_Apr2020_LDr2-8_500kb.csv\n",
      "Filtered out 65 genes based on additional criteria (no genes with SNPs in any LD with any BP loci).\n",
      "Updated intermediate least-likely genes at /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_intermediate.tsv\n",
      "Least likely genes identified and saved.\n",
      "Number of least likely genes: 571\n",
      "Number of probable genes: 145\n",
      "Number of most likely genes: 51\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step4_least_likely_gene_selection.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 6,
   "id": "e42781c2",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Using intermediate least-likely genes file: /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_intermediate.tsv\n",
      "Identified 13758 genes with at least one P<0.01 SNP in EACH Annotated_GWAS file (per-file intersection)\n",
      "Probable genes: 254 -> 145 after applying per-file P<0.01 intersection filter\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step6_identify_training_genes.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 7,
   "id": "33902de9",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Found 5116 duplicate Gene entries in merged data - aggregating numeric features by mean to collapse duplicates.\n",
      "All merged databases saved to ./example/data_preprocessing/output/all_genes_merged_all_data.csv\n",
      "Training genes counts per label:\n",
      "  least likely: 571\n",
      "  probable: 145\n",
      "  most likely: 51\n",
      "Joined data saved to ['./example/data_preprocessing/output/training_data_all_features.csv', './example/machine_learning/eda/input/training_data_all_features.csv']\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step7_merge_all_databases_and_get_training_data.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 8,
   "id": "f2f179e1",
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Using intermediate least-likely genes file: /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_intermediate.tsv\n",
      "/Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step8_downsample_least_likely_genes.py:311: UserWarning: The palette list has more values (2) than needed (1), which may not be intended.\n",
      "  sns.boxplot(data=[combined_pos['variant_count'], least_df['variant_count']], palette=['C0', 'C1'])\n",
      "Wrote diagnostic plots: /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/variant_count_hist_seed42.png /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/variant_count_box_seed42.png\n",
      "Wrote updated training_genes to ./example/data_preprocessing/input/training_genes.txt\n",
      "Wrote sampled least file: /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_sampled_seed42.tsv\n",
      "Wrote audit: /Users/hannahnicholls/GitHub/GenePrioritiser/example/data_preprocessing/input/sampling/least_likely_sampling_audit_seed42.json\n",
      "Sampled size: 51 target: 51\n",
      "Wrote training features to: ./example/data_preprocessing/output/training_data_all_features.csv\n",
      "Wrote EDA training input to: ./example/machine_learning/eda/input/training_data_all_features.csv\n"
     ]
    }
   ],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step8_downsample_least_likely_genes.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 9,
   "id": "26b264fa",
   "metadata": {},
   "outputs": [],
   "source": [
    "!python /Users/hannahnicholls/GitHub/GenePrioritiser/src/data_preprocessing/Step9_subset_genes_to_prioritise.py"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "58784add",
   "metadata": {},
   "outputs": [],
   "source": []
  }
 ],
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