From 922a8e3ca25b5218578e0133ba1fa5b176bd4a38 Mon Sep 17 00:00:00 2001 From: LuisHeinzlmeier Date: Tue, 2 Jun 2026 08:56:13 +0200 Subject: [PATCH 1/4] add a basic plot_bcv function --- .../_differential_gene_expression/_edger.py | 174 ++++++++++++++++++ 1 file changed, 174 insertions(+) diff --git a/pertpy/tools/_differential_gene_expression/_edger.py b/pertpy/tools/_differential_gene_expression/_edger.py index 63521a9b..1f2b26ab 100644 --- a/pertpy/tools/_differential_gene_expression/_edger.py +++ b/pertpy/tools/_differential_gene_expression/_edger.py @@ -1,7 +1,10 @@ from collections.abc import Sequence +import matplotlib.pyplot as plt import numpy as np import pandas as pd +from matplotlib.lines import Line2D +from matplotlib.pyplot import Figure from scipy.sparse import issparse from pertpy._logger import logger @@ -60,12 +63,183 @@ def fit(self, **kwargs): # adata, design, mask, layer logger.info("Estimating Dispersions") dge = edger.estimateDisp(dge, design=design_r) + self.dge = dge + logger.info("Fitting linear model") fit = edger.glmQLFit(dge, design=design_r, **kwargs) ro.globalenv["fit"] = fit self.fit = fit + def plot_bcv( + self, + *, + xlab: str = "Average log CPM", + ylab: str = "Biological coefficient of variation", + pch: str = "o", + cex: float = 0.2, + col_common: str = "red", + col_trend: str = "blue", + col_tagwise: str = "black", + legend: bool = True, + return_fig: bool = False, + ) -> Figure | None: + """Plot biological coefficient of variation (BCV) like edgeR::plotBCV. + + Must be called after `.fit()`. + + Args: + xlab: + X-axis label. + ylab: + Y-axis label. + pch: + Marker style (matplotlib-compatible). + cex: + Point size scaling (similar to R cex). + col_common: + Color for common dispersion line. + col_trend: + Color for trended dispersion line. + col_tagwise: + Color for tagwise dispersion points. + legend: + Whether to show legend. + return_fig: + If True, return matplotlib figure instead of showing it. + """ + if not hasattr(self, "dge"): + raise ValueError("Model not fitted yet. Call `.fit()` first.") + + # dge = self.dge + + try: + from rpy2 import robjects as ro + from rpy2.robjects import numpy2ri, pandas2ri + from rpy2.robjects.conversion import get_conversion, localconverter + from rpy2.robjects.packages import importr + + except ImportError: + raise ImportError("edger requires rpy2 to be installed.") from None + + with localconverter(get_conversion() + numpy2ri.converter): + A = np.asarray(self.dge.rx2("AveLogCPM")) + tagwise = np.asarray(self.dge.rx2("tagwise.dispersion")) if "tagwise.dispersion" in self.dge.names else None + trended = np.asarray(self.dge.rx2("trended.dispersion")) if "trended.dispersion" in self.dge.names else None + common = np.asarray(self.dge.rx2("common.dispersion"))[0] if "common.dispersion" in self.dge.names else None + + if common is None and trended is None and tagwise is None: + raise ValueError("No dispersions found in DGEList. Did you run estimateDisp()?") + + # ensure correct shapes + A = np.asarray(A) + + # ----------------------------- + # Figure setup + # ----------------------------- + fig, ax = plt.subplots(dpi=300) + + labels = [] + + # ----------------------------- + # Tagwise dispersion + # ----------------------------- + if tagwise is not None: + tagwise = np.asarray(tagwise) + + ax.scatter( + A, + np.sqrt(tagwise), + c=col_tagwise, + s=cex * 20, + marker=pch, + linewidths=0, + ) + labels.append("Tagwise") + + # ----------------------------- + # Common dispersion + # ----------------------------- + if common is not None: + common_val = float(np.asarray(common).reshape(-1)[0]) + ax.axhline( + np.sqrt(common_val), + color=col_common, + linewidth=2, + ) + labels.append("Common") + + # ----------------------------- + # Trended dispersion + # ----------------------------- + if trended is not None: + trended = np.asarray(trended) + order = np.argsort(A) + + ax.plot( + A[order], + np.sqrt(trended)[order], + color=col_trend, + linewidth=2, + ) + labels.append("Trend") + + # ----------------------------- + # Axes styling + # ----------------------------- + ax.set_xlabel(xlab) + ax.set_ylabel(ylab) + + # ----------------------------- + # Legend (R-style) + # ----------------------------- + if legend: + handles = [] + + if tagwise is not None: + handles.append( + Line2D( + [0], + [0], + marker=pch, + linestyle="", + color=col_tagwise, + label="Tagwise", + ) + ) + + if common is not None: + handles.append( + Line2D( + [0], + [0], + linestyle="-", + color=col_common, + label="Common", + ) + ) + + if trended is not None: + handles.append( + Line2D( + [0], + [0], + linestyle="-", + color=col_trend, + label="Trend", + ) + ) + + ax.legend(handles=handles, loc="upper right", frameon=True) + + plt.tight_layout() + + if return_fig: + return fig + + plt.show() + return None + def _test_single_contrast(self, contrast: Sequence[float], **kwargs) -> pd.DataFrame: # noqa: D417 """Conduct test for each contrast and return a data frame. From 33b92e6c88def4f1f6cf13ac58fec337c6e3c482 Mon Sep 17 00:00:00 2001 From: LuisHeinzlmeier Date: Wed, 3 Jun 2026 06:57:53 +0200 Subject: [PATCH 2/4] add _ensure_deps() --- .../_differential_gene_expression/_edger.py | 274 +++++++----------- 1 file changed, 105 insertions(+), 169 deletions(-) diff --git a/pertpy/tools/_differential_gene_expression/_edger.py b/pertpy/tools/_differential_gene_expression/_edger.py index 1f2b26ab..299f6c3a 100644 --- a/pertpy/tools/_differential_gene_expression/_edger.py +++ b/pertpy/tools/_differential_gene_expression/_edger.py @@ -7,6 +7,7 @@ from matplotlib.pyplot import Figure from scipy.sparse import issparse +from pertpy._doc import _doc_params, doc_common_plot_args from pertpy._logger import logger from ._base import LinearModelBase @@ -27,21 +28,9 @@ def fit(self, **kwargs): # adata, design, mask, layer Args: **kwargs: Keyword arguments specific to glmQLFit() """ - try: - from rpy2 import robjects as ro - from rpy2.robjects import numpy2ri, pandas2ri - from rpy2.robjects.conversion import get_conversion, localconverter - from rpy2.robjects.packages import importr - - except ImportError: - raise ImportError("edger requires rpy2 to be installed.") from None - - try: - edger = importr("edgeR") - except ImportError as e: - raise ImportError( - "edgeR requires a valid R installation with the following packages:\nedgeR, BiocParallel, RhpcBLASctl" - ) from e + ro, numpy2ri, pandas2ri, get_conversion, localconverter, edger = self._ensure_deps( + "ro", "numpy2ri", "pandas2ri", "get_conversion", "localconverter", "edger" + ) # Convert dataframe with localconverter(get_conversion() + numpy2ri.converter): @@ -71,165 +60,82 @@ def fit(self, **kwargs): # adata, design, mask, layer ro.globalenv["fit"] = fit self.fit = fit - def plot_bcv( + @_doc_params(common_plot_args=doc_common_plot_args) + def plot_bcv( # pragma: no cover # noqa: D417 self, *, - xlab: str = "Average log CPM", - ylab: str = "Biological coefficient of variation", - pch: str = "o", - cex: float = 0.2, - col_common: str = "red", - col_trend: str = "blue", - col_tagwise: str = "black", + xlabel: str | None = "Average log CPM", + ylabel: str | None = "Biological coefficient of variation", + marker: str = "o", + point_size: float = 0.2, + common_col: str = "red", + trend_col: str = "blue", + tagwise_col: str = "black", legend: bool = True, return_fig: bool = False, + **kwargs, ) -> Figure | None: """Plot biological coefficient of variation (BCV) like edgeR::plotBCV. - Must be called after `.fit()`. - Args: - xlab: - X-axis label. - ylab: - Y-axis label. - pch: - Marker style (matplotlib-compatible). - cex: - Point size scaling (similar to R cex). - col_common: - Color for common dispersion line. - col_trend: - Color for trended dispersion line. - col_tagwise: - Color for tagwise dispersion points. - legend: - Whether to show legend. - return_fig: - If True, return matplotlib figure instead of showing it. + xlabel: Label for the x-axis (default: "Average log CPM"). + ylabel: Label for the y-axis (default: "Biological coefficient of variation"). + marker: Marker style. + point_size: Scaling factor for point sizes. + common_col: Color for common dispersion line. + trend_col: Color for trended dispersion line. + tagwise_col: Color for tagwise dispersion points. + legend: Whether to draw a legend. + {common_plot_args} + **kwargs: Additional arguments for ax.scatter and ax.axhline. """ + # TODO if not hasattr(self, "dge"): raise ValueError("Model not fitted yet. Call `.fit()` first.") - # dge = self.dge - - try: - from rpy2 import robjects as ro - from rpy2.robjects import numpy2ri, pandas2ri - from rpy2.robjects.conversion import get_conversion, localconverter - from rpy2.robjects.packages import importr - - except ImportError: - raise ImportError("edger requires rpy2 to be installed.") from None + numpy2ri, get_conversion, localconverter = self._ensure_deps("numpy2ri", "get_conversion", "localconverter") with localconverter(get_conversion() + numpy2ri.converter): A = np.asarray(self.dge.rx2("AveLogCPM")) - tagwise = np.asarray(self.dge.rx2("tagwise.dispersion")) if "tagwise.dispersion" in self.dge.names else None - trended = np.asarray(self.dge.rx2("trended.dispersion")) if "trended.dispersion" in self.dge.names else None - common = np.asarray(self.dge.rx2("common.dispersion"))[0] if "common.dispersion" in self.dge.names else None + tagwise = np.asarray(self.dge.rx2("tagwise.dispersion")) + common = float(self.dge.rx2("common.dispersion")[0]) + trended = np.asarray(self.dge.rx2("trended.dispersion")) - if common is None and trended is None and tagwise is None: - raise ValueError("No dispersions found in DGEList. Did you run estimateDisp()?") + fig, ax = plt.subplots(dpi=300) - # ensure correct shapes - A = np.asarray(A) + ax.scatter( + A, + np.sqrt(tagwise), + c=tagwise_col, + s=point_size * 20, + marker=marker, + linewidths=0, + ) - # ----------------------------- - # Figure setup - # ----------------------------- - fig, ax = plt.subplots(dpi=300) + ax.axhline( + np.sqrt(common), + color=common_col, + linewidth=2, + ) - labels = [] - - # ----------------------------- - # Tagwise dispersion - # ----------------------------- - if tagwise is not None: - tagwise = np.asarray(tagwise) - - ax.scatter( - A, - np.sqrt(tagwise), - c=col_tagwise, - s=cex * 20, - marker=pch, - linewidths=0, - ) - labels.append("Tagwise") - - # ----------------------------- - # Common dispersion - # ----------------------------- - if common is not None: - common_val = float(np.asarray(common).reshape(-1)[0]) - ax.axhline( - np.sqrt(common_val), - color=col_common, - linewidth=2, - ) - labels.append("Common") - - # ----------------------------- - # Trended dispersion - # ----------------------------- - if trended is not None: - trended = np.asarray(trended) - order = np.argsort(A) - - ax.plot( - A[order], - np.sqrt(trended)[order], - color=col_trend, - linewidth=2, - ) - labels.append("Trend") - - # ----------------------------- - # Axes styling - # ----------------------------- - ax.set_xlabel(xlab) - ax.set_ylabel(ylab) - - # ----------------------------- - # Legend (R-style) - # ----------------------------- - if legend: - handles = [] - - if tagwise is not None: - handles.append( - Line2D( - [0], - [0], - marker=pch, - linestyle="", - color=col_tagwise, - label="Tagwise", - ) - ) - - if common is not None: - handles.append( - Line2D( - [0], - [0], - linestyle="-", - color=col_common, - label="Common", - ) - ) - - if trended is not None: - handles.append( - Line2D( - [0], - [0], - linestyle="-", - color=col_trend, - label="Trend", - ) - ) + order = np.argsort(A) + ax.plot( + A[order], + np.sqrt(trended)[order], + color=trend_col, + linewidth=2, + ) + + ax.set_xlabel(xlabel) + ax.set_ylabel(ylabel) + + if legend: + handles = [ + Line2D([0], [0], marker=marker, linestyle="", color=tagwise_col, label="Tagwise"), + Line2D([0], [0], linestyle="-", color=common_col, label="Common"), + Line2D([0], [0], linestyle="-", color=trend_col, label="Trend"), + ] ax.legend(handles=handles, loc="upper right", frameon=True) plt.tight_layout() @@ -255,21 +161,9 @@ def _test_single_contrast(self, contrast: Sequence[float], **kwargs) -> pd.DataF # parse **kwargs to R function # Fix mask for .fit() - try: - from rpy2 import robjects as ro - from rpy2.robjects import numpy2ri, pandas2ri - from rpy2.robjects.conversion import get_conversion, localconverter - from rpy2.robjects.packages import importr - - except ImportError: - raise ImportError("edger requires rpy2 to be installed.") from None - - try: - importr("edgeR") - except ImportError: - raise ImportError( - "edgeR requires a valid R installation with the following packages: edgeR, BiocParallel, RhpcBLASctl" - ) from None + ro, numpy2ri, pandas2ri, get_conversion, localconverter = self._ensure_deps( + "ro", "numpy2ri", "pandas2ri", "get_conversion", "localconverter" + ) # Convert vector to R, which drops a category like `self.design_matrix` to use the intercept for the left out. with localconverter(get_conversion() + numpy2ri.converter) as cv: @@ -300,3 +194,45 @@ def _test_single_contrast(self, contrast: Sequence[float], **kwargs) -> pd.DataF de_res = de_res.reset_index() return de_res.rename(columns={"PValue": "p_value", "logFC": "log_fc", "FDR": "adj_p_value"}) + + def _ensure_deps(self, *names): + """Lazy loader for rpy2 objects with per-instance caching. + + Example: + ro, numpy2ri, edger = self._ensure_deps("ro", "numpy2ri", "edger") + """ + if not hasattr(self, "_imports_cache"): + try: + from rpy2 import robjects as ro + from rpy2.robjects import numpy2ri, pandas2ri + from rpy2.robjects.conversion import get_conversion, localconverter + from rpy2.robjects.packages import importr + + except ImportError: + raise ImportError("edger requires rpy2 to be installed.") from None + + try: + edger = importr("edgeR") + except ImportError as e: + raise ImportError( + "edgeR requires a valid R installation with the following packages:\nedgeR, BiocParallel, RhpcBLASctl" + ) from e + + self._imports_cache = { + "ro": ro, + "numpy2ri": numpy2ri, + "pandas2ri": pandas2ri, + "get_conversion": get_conversion, + "localconverter": localconverter, + "edger": edger, + } + + results = {} + + for name in names: + if name in self._imports_cache: + results[name] = self._imports_cache[name] + else: + raise KeyError(f"Unknown import request: '{name}'") + + return tuple(results[name] for name in names) From 5e7e0f257fc8b4042e1fbc7dedf237f69519e30f Mon Sep 17 00:00:00 2001 From: LuisHeinzlmeier Date: Wed, 3 Jun 2026 07:19:51 +0200 Subject: [PATCH 3/4] finalize --- .../_differential_gene_expression/_edger.py | 83 +++++++++---------- 1 file changed, 37 insertions(+), 46 deletions(-) diff --git a/pertpy/tools/_differential_gene_expression/_edger.py b/pertpy/tools/_differential_gene_expression/_edger.py index 299f6c3a..6e82dd92 100644 --- a/pertpy/tools/_differential_gene_expression/_edger.py +++ b/pertpy/tools/_differential_gene_expression/_edger.py @@ -28,34 +28,13 @@ def fit(self, **kwargs): # adata, design, mask, layer Args: **kwargs: Keyword arguments specific to glmQLFit() """ - ro, numpy2ri, pandas2ri, get_conversion, localconverter, edger = self._ensure_deps( - "ro", "numpy2ri", "pandas2ri", "get_conversion", "localconverter", "edger" - ) + ro, edger = self._ensure_deps("ro", "edger") - # Convert dataframe - with localconverter(get_conversion() + numpy2ri.converter): - expr = self.adata.X if self.layer is None else self.adata.layers[self.layer] - expr = expr.T.toarray() if issparse(expr) else expr.T - - with localconverter(get_conversion() + pandas2ri.converter) as cv: - expr_r = cv.py2rpy(pd.DataFrame(expr, index=self.adata.var_names, columns=self.adata.obs_names)) - samples_r = cv.py2rpy(self.adata.obs) - - dge = edger.DGEList(counts=expr_r, samples=samples_r) - - logger.info("Calculating NormFactors") - dge = edger.calcNormFactors(dge) - - with localconverter(get_conversion() + numpy2ri.converter) as cv: - design_r = cv.py2rpy(self.design.values) - - logger.info("Estimating Dispersions") - dge = edger.estimateDisp(dge, design=design_r) - - self.dge = dge + if not hasattr(self, "dge") or not hasattr(self, "design_r"): + self._prepare_dge() logger.info("Fitting linear model") - fit = edger.glmQLFit(dge, design=design_r, **kwargs) + fit = edger.glmQLFit(self.dge, design=self.design_r, **kwargs) ro.globalenv["fit"] = fit self.fit = fit @@ -89,9 +68,8 @@ def plot_bcv( # pragma: no cover # noqa: D417 {common_plot_args} **kwargs: Additional arguments for ax.scatter and ax.axhline. """ - # TODO if not hasattr(self, "dge"): - raise ValueError("Model not fitted yet. Call `.fit()` first.") + self._prepare_dge() numpy2ri, get_conversion, localconverter = self._ensure_deps("numpy2ri", "get_conversion", "localconverter") @@ -103,29 +81,13 @@ def plot_bcv( # pragma: no cover # noqa: D417 fig, ax = plt.subplots(dpi=300) - ax.scatter( - A, - np.sqrt(tagwise), - c=tagwise_col, - s=point_size * 20, - marker=marker, - linewidths=0, - ) + ax.scatter(A, np.sqrt(tagwise), c=tagwise_col, s=point_size * 20, marker=marker, linewidths=0, **kwargs) - ax.axhline( - np.sqrt(common), - color=common_col, - linewidth=2, - ) + ax.axhline(np.sqrt(common), color=common_col, linewidth=2, **kwargs) order = np.argsort(A) - ax.plot( - A[order], - np.sqrt(trended)[order], - color=trend_col, - linewidth=2, - ) + ax.plot(A[order], np.sqrt(trended)[order], color=trend_col, linewidth=2, **kwargs) ax.set_xlabel(xlabel) ax.set_ylabel(ylabel) @@ -146,6 +108,35 @@ def plot_bcv( # pragma: no cover # noqa: D417 plt.show() return None + def _prepare_dge(self) -> None: + """Create DGEList, calculate normalization factors, and estimate dispersions.""" + numpy2ri, pandas2ri, get_conversion, localconverter, edger = self._ensure_deps( + "numpy2ri", "pandas2ri", "get_conversion", "localconverter", "edger" + ) + + # Convert dataframe + with localconverter(get_conversion() + numpy2ri.converter): + expr = self.adata.X if self.layer is None else self.adata.layers[self.layer] + expr = expr.T.toarray() if issparse(expr) else expr.T + + with localconverter(get_conversion() + pandas2ri.converter) as cv: + expr_r = cv.py2rpy(pd.DataFrame(expr, index=self.adata.var_names, columns=self.adata.obs_names)) + samples_r = cv.py2rpy(self.adata.obs) + + dge = edger.DGEList(counts=expr_r, samples=samples_r) + + logger.info("Calculating NormFactors") + dge = edger.calcNormFactors(dge) + + with localconverter(get_conversion() + numpy2ri.converter) as cv: + design_r = cv.py2rpy(self.design.values) + + logger.info("Estimating Dispersions") + dge = edger.estimateDisp(dge, design=design_r) + + self.dge = dge + self.design_r = design_r + def _test_single_contrast(self, contrast: Sequence[float], **kwargs) -> pd.DataFrame: # noqa: D417 """Conduct test for each contrast and return a data frame. 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adata.layers["counts"] = adata.X.copy() + >>> pdata = dc.pp.pseudobulk(adata, sample_col="Patient", groups_col="Cluster", layer="counts", mode="sum") + >>> dc.pp.filter_samples(pdata, inplace=True) + >>> edgr = pt.tl.EdgeR(pdata, design="~Efficacy+Treatment") + >>> edgr.plot_bcv() + + Preview: + .. image:: /_static/docstring_previews/de_plot_bcv.png """ if not hasattr(self, "dge"): self._prepare_dge()