import logging from typing import Any import anndata as ad import numpy as np import polars as pl from numpy.typing import NDArray from pdex import parallel_differential_expression from scipy.sparse import issparse from ._evaluator import _build_pdex_kwargs, _convert_to_normlog logger = logging.getLogger(__name__) def build_base_mean_adata( adata: ad.AnnData | str, counts_df: pl.DataFrame | str | None = None, pert_col: str = "target_gene", control_pert: str = "non-targeting", counts_col: str = "n_cells", as_delta: bool = False, allow_discrete: bool = False, output_path: str | None = None, output_de_path: str | None = None, batch_size: int = 1000, num_threads: int = 1, de_method: str = "wilcoxon", pdex_kwargs: dict[str, Any] = {}, ) -> ad.AnnData: if isinstance(adata, str): logger.info(f"Reading adata from path: {adata}") adata = ad.read_h5ad(adata) # Convert to normalized log space if necessary _convert_to_normlog(adata=adata, allow_discrete=allow_discrete) counts = ( _load_counts_df( counts_df=counts_df, pert_col=pert_col, control_pert=control_pert, counts_col=counts_col, ) if counts_df is not None else _build_counts_df_from_adata( adata=adata, pert_col=pert_col, control_pert=control_pert, counts_col=counts_col, ) ) baseline = _build_pert_baseline( adata=adata, pert_col=pert_col, control_pert=control_pert, as_delta=as_delta ) obs = ( counts.select([pl.col(pert_col).repeat_by(counts_col)]) .explode(pert_col) .to_pandas() ) obs.index = obs.index.astype(str).str.replace("^", "p.", regex=True) logger.info("Assembling baseline adata from perturbation mean") baseline_adata = ad.AnnData( X=np.full( (int(counts[counts_col].sum()), baseline.size), baseline, ), var=adata.var, obs=obs, ) logger.info("Concatenating baseline adata with controls from original adata") baseline_adata = ad.concat( [baseline_adata, adata[adata.obs[pert_col] == control_pert]] ) if output_path is not None: logger.info(f"Saving baseline data to {output_path}") baseline_adata.write_h5ad(output_path) if output_de_path is not None: logger.info("Calculating differential expression") pdex_kwargs = _build_pdex_kwargs( groupby_key=pert_col, reference=control_pert, num_workers=num_threads, metric=de_method, batch_size=batch_size, allow_discrete=allow_discrete, pdex_kwargs=pdex_kwargs, ) frame = parallel_differential_expression( adata=baseline_adata, **pdex_kwargs, ) logger.info(f"Saving differential expression results to {output_de_path}") frame.write_csv(output_de_path) return baseline_adata def _load_counts_df( counts_df: pl.DataFrame | str, pert_col: str = "target_gene", control_pert: str = "non-targeting", counts_col: str = "n_cells", ) -> pl.DataFrame: if isinstance(counts_df, str): logger.info(f"Loading counts from {counts_df}") counts_df = pl.read_csv(counts_df) if pert_col not in counts_df.columns: raise ValueError( f"Column '{pert_col}' not found in counts_df: {counts_df.columns}" ) if counts_col not in counts_df.columns: raise ValueError( f"Column '{counts_col}' not found in counts_df: {counts_df.columns}" ) logger.info(f"Filtering out counts from {control_pert}") return counts_df.filter( pl.col(pert_col) != control_pert # drop control pert ) def _build_counts_df_from_adata( adata: ad.AnnData, pert_col: str = "target_gene", control_pert: str = "non-targeting", counts_col: str = "n_cells", ) -> pl.DataFrame: if pert_col not in adata.obs.columns: raise ValueError( f"Column '{pert_col}' not found in adata.obs: {adata.obs.columns}" ) if control_pert not in adata.obs[pert_col].unique(): raise ValueError( f"Control pert '{control_pert}' not found in adata.obs[{pert_col}]: {adata.obs[pert_col].unique()}" ) logger.info("Building counts DataFrame from adata") return ( pl.DataFrame(adata.obs) .group_by(pert_col) .len() .rename({"len": counts_col}) .filter(pl.col(pert_col) != control_pert) ) def _build_pert_baseline( adata: ad.AnnData, pert_col: str = "target_gene", control_pert: str = "non-targeting", as_delta: bool = False, ) -> NDArray[np.float64]: if pert_col not in adata.obs.columns: raise ValueError( f"Column '{pert_col}' not found in adata.obs: {adata.obs.columns}" ) unique_perts = adata.obs[pert_col].unique() if control_pert not in unique_perts: raise ValueError( f"Control pert '{control_pert}' not found in unique_perts: {unique_perts}" ) logger.info("Building perturbation-level means") pert_means = ( pl.DataFrame( adata.X if not issparse(adata.X) else adata.X.toarray() # type: ignore ) .with_columns(pl.Series(pert_col, adata.obs[pert_col])) .group_by(pert_col) .mean() ) names = pert_means.drop_in_place(pert_col).to_numpy() pert_mask = names != control_pert pert_matrix = pert_means.to_numpy() if as_delta: logger.info("Calculating delta from control means") delta = pert_matrix[pert_mask] - pert_matrix[~pert_mask] logger.info("Calculating mean delta") mean_delta = delta.mean(axis=0) return mean_delta else: logger.info("Calculating mean of perturbation-level means") mean_pert = pert_matrix.mean(axis=0) return mean_pert