From 6f9887a086554621d8834b6f10035f83b158374c Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Wed, 8 Jul 2020 16:06:02 -0700 Subject: [PATCH 01/10] Started trajectory --- genetools/__init__.py | 4 +- genetools/plots.py | 239 ++++++++++-------- genetools/trajectory.py | 530 +++++++++++++++++++++++++++++++++++++++ tests/test_trajectory.py | 77 ++++++ 4 files changed, 742 insertions(+), 108 deletions(-) create mode 100644 genetools/trajectory.py create mode 100644 tests/test_trajectory.py diff --git a/genetools/__init__.py b/genetools/__init__.py index 62b817a..127cfd2 100644 --- a/genetools/__init__.py +++ b/genetools/__init__.py @@ -5,6 +5,6 @@ __version__ = "0.4.0" # We want genetools.[submodule].[function] to be accessible simply by importing genetools, without having to import genetools.[submodule] -from . import helpers, plots, scanpy_helpers, stats +from . import helpers, plots, scanpy_helpers, stats, trajectory -__all__ = ["helpers", "plots", "scanpy_helpers", "stats"] +__all__ = ["helpers", "plots", "scanpy_helpers", "stats", "trajectory"] diff --git a/genetools/plots.py b/genetools/plots.py index 0c091a6..1f292f4 100644 --- a/genetools/plots.py +++ b/genetools/plots.py @@ -29,6 +29,11 @@ def umap_scatter( label_color="k", label_alpha=0.5, label_size=20, + highlight_cell_names=None, + highlight_marker_size=30, + highlight_marker_color="r", + highlight_zorder=5, + highlight_marker_style="s", ): """Simple umap scatter plot, with legend outside figure. @@ -68,6 +73,16 @@ def umap_scatter( :type label_alpha: float, optional :param label_size: size of cluster labels, defaults to 20 :type label_size: int, optional + :param highlight_cell_names: [description] + :type highlight_cell_names: [type] + :param highlight_marker_size: [description], defaults to 30 + :type highlight_marker_size: int, optional + :param highlight_marker_color: [description], defaults to 'r' + :type highlight_marker_color: str, optional + :param highlight_zorder: [description], defaults to 5 + :type highlight_zorder: int, optional + :param highlight_marker_style: [description], defaults to 's' + :type highlight_marker_style: str, optional :return: matplotlib figure and axes :rtype: (matplotlib.Figure, matplotlib.Axes) """ @@ -125,6 +140,17 @@ def umap_scatter( zorder=label_z_order, ) + # overlay: highlight cells + if highlight_cell_names is not None: + plt.scatter( + data.iloc[highlight_cell_names][umap_1_key], + data.iloc[highlight_cell_names][umap_2_key], + s=highlight_marker_size, + c=highlight_marker_color, + zorder=highlight_zorder, + marker=highlight_marker_style, + ) + sns.despine(ax=ax) plt.tight_layout() @@ -260,112 +286,113 @@ def horizontal_stacked_bar_plot( #### -# def stacked_density_plot( -# data, -# row_var, -# hue_var, -# value_var, -# col_var=None, -# overlap=False, -# suptitle=None, -# figsize=None, -# hue_order=None, -# row_order=None, -# palette=None, -# ): -# """ -# Multiple density plot. -# Adapted from old work at https://github.com/hammerlab/infino/blob/develop/analyze_cut.py#L912 - -# For row_order, consider row_order=reversed(list(range(data.ylevel.values.max()+1))) -# """ - -# with sns.plotting_context("notebook"): -# with sns.axes_style("white", rc={"axes.facecolor": (0, 0, 0, 0)}): -# g = sns.FacetGrid( -# data, -# row=row_var, -# hue=hue_var, -# col=col_var, -# row_order=row_order, -# hue_order=hue_order, -# aspect=15, -# height=0.5, -# palette=palette, -# sharey=False, # important -- they don't share y ranges. -# ) - -# ## Draw the densities in a few steps -# # this is the shaded area -# g.map(sns.kdeplot, value_var, clip_on=False, shade=True, alpha=0.8, lw=2) - -# # this is the dividing horizontal line -# g.map(plt.axhline, y=0, lw=2, clip_on=False, ls="dashed") - -# ### Add label for each facet. - -# def label(**kwargs): -# """ -# kwargs is e.g.: {'color': (0.4918017777777778, 0.25275644444444445, 0.3333333333333333), 'label': 'Name of the row'} -# """ -# color = kwargs["color"] -# label = kwargs["label"] -# ax = plt.gca() # map() changes current axis repeatedly -# # x=1 if plot_on_right else 0; ha="right" if plot_on_right else "left", -# ax.text( -# 1.25, -# 0.5, -# label, -# # fontweight="bold", -# color=color, -# # ha="right", -# ha="left", -# va="center", -# transform=ax.transAxes, -# fontsize="x-small", -# # fontsize='x-large', #15, -# # bbox=dict(facecolor='yellow', alpha=0.3) -# ) - -# g.map(label) - -# ## Beautify the plot. -# g.set(xlim=(-0.01, 1.01)) -# # seems to do the trick along with sharey=False -# g.set(ylim=(0, None)) - -# # Some `subplots_adjust` line is necessary. without this, nothing appears -# if not overlap: -# g.fig.subplots_adjust(hspace=0) - -# # Remove axes details that don't play will with overlap -# g.set_titles("") -# # g.set_titles(col_template="{col_name}", row_template="") -# g.set(yticks=[], ylabel="") -# g.despine(bottom=True, left=True) - -# # fix x axis -# g.set_xlabels("Pseudotime") - -# # resize -# if figsize: -# g.fig.set_size_inches(figsize[0], figsize[1]) -# else: -# cur_size = g.fig.get_size_inches() -# increase_vertical = 3 # 7 #4 # 3 -# g.fig.set_size_inches(cur_size[0], cur_size[1] + increase_vertical) - -# if suptitle is not None: -# g.fig.suptitle(suptitle, fontsize="medium") - -# # tighten -# g.fig.tight_layout() - -# # then reoverlap -# if overlap: -# g.fig.subplots_adjust(hspace=-0.1) - -# return g, g.fig +def stacked_density_plot( + data, + row_var, + hue_var, + value_var, + xlabel, + col_var=None, + overlap=False, + suptitle=None, + figsize=None, + hue_order=None, + row_order=None, + palette=None, +): + """ + Multiple density plot. + Adapted from old work at https://github.com/hammerlab/infino/blob/develop/analyze_cut.py#L912 + + For row_order, consider row_order=reversed(list(range(data.ylevel.values.max()+1))) + """ + + with sns.plotting_context("notebook"): + with sns.axes_style("white", rc={"axes.facecolor": (0, 0, 0, 0)}): + g = sns.FacetGrid( + data, + row=row_var, + hue=hue_var, + col=col_var, + row_order=row_order, + hue_order=hue_order, + aspect=15, + height=0.5, + palette=palette, + sharey=False, # important -- they don't share y ranges. + ) + + ## Draw the densities in a few steps + # this is the shaded area + g.map(sns.kdeplot, value_var, clip_on=False, shade=True, alpha=0.8, lw=2) + + # this is the dividing horizontal line + g.map(plt.axhline, y=0, lw=2, clip_on=False, ls="dashed") + + ### Add label for each facet. + + def label(**kwargs): + """ + kwargs is e.g.: {'color': (0.4918017777777778, 0.25275644444444445, 0.3333333333333333), 'label': 'Name of the row'} + """ + color = kwargs["color"] + label = kwargs["label"] + ax = plt.gca() # map() changes current axis repeatedly + # x=1 if plot_on_right else 0; ha="right" if plot_on_right else "left", + ax.text( + 1.25, + 0.5, + label, + # fontweight="bold", + color=color, + # ha="right", + ha="left", + va="center", + transform=ax.transAxes, + fontsize="x-small", + # fontsize='x-large', #15, + # bbox=dict(facecolor='yellow', alpha=0.3) + ) + + g.map(label) + + ## Beautify the plot. + g.set(xlim=(-0.01, 1.01)) + # seems to do the trick along with sharey=False + g.set(ylim=(0, None)) + + # Some `subplots_adjust` line is necessary. without this, nothing appears + if not overlap: + g.fig.subplots_adjust(hspace=0) + + # Remove axes details that don't play will with overlap + g.set_titles("") + # g.set_titles(col_template="{col_name}", row_template="") + g.set(yticks=[], ylabel="") + g.despine(bottom=True, left=True) + + # fix x axis + g.set_xlabels(xlabel) + + # resize + if figsize: + g.fig.set_size_inches(figsize[0], figsize[1]) + else: + cur_size = g.fig.get_size_inches() + increase_vertical = 3 # 7 #4 # 3 + g.fig.set_size_inches(cur_size[0], cur_size[1] + increase_vertical) + + if suptitle is not None: + g.fig.suptitle(suptitle, fontsize="medium") + + # tighten + g.fig.tight_layout() + + # then reoverlap + if overlap: + g.fig.subplots_adjust(hspace=-0.1) + + return g.fig # TODO: density umap plot diff --git a/genetools/trajectory.py b/genetools/trajectory.py new file mode 100644 index 0000000..e60dc14 --- /dev/null +++ b/genetools/trajectory.py @@ -0,0 +1,530 @@ +# run on all +import numpy as np +import pandas as pd +from genetools import plots, stats +import matplotlib.pyplot as plt +import seaborn as sns + + +def monte_carlo_pseudotime(adata_input, roots, id_vars): + import scanpy as sc + + """[summary] + + :param adata_input: [description] + :type adata_input: [type] + :param roots: [description] + :type roots: [type] + :param id_vars: Identifier variables in anndata obs to uniquely identify a cell. Will be passed through to output. + :type id_vars: [type] + :return: [description] + :rtype: [type] + """ + # TODO: can we avoid passing in id_vars, and just set 'full_barcode' from obs_names + + adata = adata_input.copy() + + if "root_cell" in id_vars or "pseudotime" in id_vars: + raise ValueError("Cannot include 'root_cell' or 'pseudotime' in id_vars") + + # root cell names + col_names = [] + + # each entry is one trajectory from one root cell: a Series, values are the pseudotime values per-cell, and name is the root cell + pseudotime_vals = [] + + for root_cell in roots: + # Set root cell + adata.uns["iroot"] = np.where(adata.obs.index == root_cell)[0][0] + + # Run DPT. Scanpy stores output in obs 'dpt_pseudotime + sc.tl.dpt(adata, n_dcs=min(adata.obsm["X_diffmap"].shape[1], 10), copy=False) + + # Save this trajectory + output = adata.obs["dpt_pseudotime"].copy() + output.name = root_cell + col_names.append(output.name) + pseudotime_vals.append(output) + + # percentile normalize each pseudotime trajectory + pseudotime_vals = pd.concat(pseudotime_vals, axis=1) + pseudotime_vals_percentile_normalized = pseudotime_vals.apply( + stats.percentile_normalize, axis=0 + ) + if pseudotime_vals_percentile_normalized.shape != pseudotime_vals.shape: + raise ValueError() + + # Create dataframe where each row is a cell's value in a trajectory from a specific root cell. + # i.e. convert to format: one value per cell, times # experiments + df = pd.concat([adata_input.obs] + pseudotime_vals_percentile_normalized, axis=1) + df = pd.melt( + df.reset_index(), + id_vars=id_vars, + value_vars=col_names, + var_name="root_cell", + value_name="pseudotime", + ) + + if df.shape[0] != adata_input.obs.shape[0] * len(roots): + raise ValueError( + "Should have one value per cell, times the number of experiments" + ) + + return df + + +def choose_roots(adata, n_roots, cluster_key, cluster_names): + """[summary] + + + + :param n_roots: [description] + :type n_roots: [type] + :param cluster_key: [description] + :type cluster_key: [type] + :param cluster_names: [description] + :type cluster_names: [type] + :return: [description] + :rtype: [type] + """ + return ( + adata.obs[adata.obs[cluster_key].isin(cluster_names)] + .index.to_series() + .sample(n_roots) + .values + ) + + +def get_end_points(trajectory_df, id_vars): + """[summary] + + :param trajectory_df: [description] + :type trajectory_df: [type] + :param id_vars: [description] + :type id_vars: [type] + :return: [description] + :rtype: [type] + """ + return trajectory_df.loc[ + trajectory_df.groupby("root_cell", sort=False)["pseudotime"].idxmax() + ][id_vars] + + +def stochasticity( + trajectory_df, + xlabel="Pseudotime", + ylabel="Number of unique barcodes in bin", + title="How stochastic are different parts of the trajectory?", +): + """[summary] + + :param trajectory_df: [description] + :type trajectory_df: [type] + :param xlabel: [description], defaults to "Pseudotime" + :type xlabel: str, optional + :param ylabel: [description], defaults to "Number of unique barcodes in bin" + :type ylabel: str, optional + :param title: [description], defaults to "How stochastic are different parts of the trajectory?" + :type title: str, optional + :return: [description] + :rtype: [type] + """ + df = trajectory_df.copy() + + # make bins + pt_bins = np.histogram_bin_edges( + df["pseudotime"], bins=50, range=(0, 1) + ) # try bins='auto' + + # cut data into bins + df["bin_id"] = pd.cut(df["pseudotime"], pt_bins) # alternative: np.digitize + + # change bin ID to be left-offset of bin instead + df["bin_id"] = df["bin_id"].apply(lambda interval: interval.left) + + # Histogram: number of values in each bin + # for this analysis, we want this to be roughly uniform. + # that's why we use percentile normalized pseudotime values above! otherwise it won't be uniform. + # df["bin_id"].hist() + + # group by bin ID, count unique barcodes in that bin + cells_by_bin = df.groupby("bin_id", sort=False)["full_barcode"].nunique() + + fig = plt.figure() + plt.scatter(cells_by_bin.index, cells_by_bin.values) + if xlabel is not None: + plt.xlabel(xlabel) + if ylabel is not None: + plt.ylabel(ylabel) + if title is not None: + plt.title(title) + return fig + + +def mean_order(trajectory_df, barcode_key): + """[summary] + ## Ensemble: Mean trajectory + + trajectories have different linear scales: + + - so percentile normalize each trajectory first: replace start value by 0, end by 1, median by 0.5. now they have same scale, so averaging sensible (already done) + - make sure nans→0s are being treated carefully, i.e. they are not being ranked ordinally. the pre-existing 0s (start cells) should be normalized to 0. + - actually there are no nans above. + + In tests: + # make sure concattable to obs + assert all(mean_trajectory.index == adata.obs.index) + + # adata.obs["mean_trajectory"] = ensemble_pt_trajectories( + # adata, col_names, pseudotime_vals, roots + # ) + + :param trajectory_df: [description] + :type trajectory_df: [type] + :param barcode_key: [description] + :type barcode_key: [type] + :return: [description] + :rtype: [type] + """ + if any(trajectory_df["pseudotime"].isna()): + raise ValueError("Some cells are unreachable (NaN pseudotimes)") + + # Take each cell's mean across all trajectories + # "project cells onto this ensemble medoid trajectory" + mean_trajectory = trajectory_df.groupby([barcode_key], sort=False)[ + "pseudotime" + ].mean() + if any(mean_trajectory.isna()): + raise ValueError() + + # renormalize + mean_trajectory = pd.Series( + stats.percentile_normalize(mean_trajectory), index=mean_trajectory.index + ) + + return mean_trajectory + + +def spectral_order( + trajectory_df, + root_cell_key, + barcode_key, + n_trajectories_sample=None, + n_cells_sample=None, +): + """[summary] + Expect percent normalized input. + + Use a kNN graph from scanpy instead of computing a dense similarity matrix: + + 1. Make P, the cells-x-cells binary comparisons matrix of 1s and -1s based on cells beating each other in pseudotime trajectory, averaged over pseudotime trajectories. + - Subsample cells at random. + - Subsample matches at random. + - At a point the basic order will pop out, no need to be perfect. + 2. Normalize each row to have same Euclidean norm + 3. Create Anndata from P. (View P as the usual cells x features matrix, where each cell is featurized in terms of the others.) + 4. Run scanpy `sc.pp.neighbors`. Now you have a kNN graph, like a compressed version of the full similarity graph above. + 5. Laplacian on `adata.uns['neighbors']['connectivities']`, the adjacency matrix of the kNN graph. + 6. 2nd smallest eigenvector + + Previously, we were creating dense similarity matrix, where the similarity measure between two cells `i, j` is the sum over all other cells `k` of $1 - abs(P_{i,j} - P_{j,k})$. The intuition is that this gives you the ranking similarity between any two cells by comparing their probabilities of being ranked higher than other reference cells. Now are are simplifying how to create this similarity matrix. + + + :param trajectory_df: [description] + :type trajectory_df: [type] + :param root_cell_key: [description] + :type root_cell_key: [type] + :param barcode_key: [description] + :type barcode_key: [type] + :param n_trajectories_sample: [description], defaults to None + :type n_trajectories_sample: [type], optional + :param n_cells_sample: [description], defaults to None + :type n_cells_sample: [type], optional + :raises ValueError: [description] + :return: [description] + :rtype: [type] + """ + + # TODO: confirm percentile normalized input + + import scipy.sparse + from scipy.sparse.csgraph import laplacian + from scipy.sparse.linalg import eigsh + from sklearn import preprocessing + import anndata + import scanpy as sc + + # Legacy code from when we passed around arrays of individual trajectories + # (We might bring that API back): + + # pseudotime_ranks = pd.concat(pseudotime_vals, axis=1) + # pseudotime_ranks = pseudotime_ranks.apply(percentile_normalize, axis=0) + # pseudotime_ranks = pseudotime_ranks.T # n_experiments x n_cells, percentile normalized + + # print(pseudotime_ranks.shape) + # pseudotime_ranks + + # cells_sample = np.random.choice( + # pseudotime_ranks.columns, size=n_cells_sample, replace=False + # ) + # cells_sample[:5] + + # matches_sample = np.random.choice( + # pseudotime_ranks.index, size=n_trajectories_sample, replace=False + # ) + # matches_sample[:5] + + # pseudotime_ranks_subsample = pseudotime_ranks.loc[matches_sample][cells_sample] + # pseudotime_ranks_subsample.shape, pseudotime_ranks.shape + + if not n_cells_sample: + # default to 20% of cells + n_cells_sample = int(trajectory_df[barcode_key].nunique() * 0.2) + if not n_trajectories_sample: + # default to 10% of trajectories + n_trajectories_sample = int(trajectory_df[root_cell_key].nunique() * 0.1) + + cells_sample = np.random.choice( + trajectory_df[barcode_key].unique(), size=n_cells_sample, replace=False + ) + matches_sample = np.random.choice( + trajectory_df[root_cell_key].unique(), size=n_trajectories_sample, replace=False + ) + # n_experiments x n_cells, percentile normalized + pseudotime_ranks_subsample = trajectory_df[ + (trajectory_df[root_cell_key].isin(matches_sample)) + & (trajectory_df[barcode_key].isin(cells_sample)) + ].pivot(index=root_cell_key, columns=barcode_key, values="pseudotime") + + # make P, the transition matrix + # # number of matches, as in a tournament where each match = all players compete against each other + # n_matches = pseudotime_ranks_subsample.shape[0] + # # number of players in the tournament + # n_cells = pseudotime_ranks_subsample.shape[1] + + matches = scipy.sparse.csr_matrix( + (pseudotime_ranks_subsample.shape[1], pseudotime_ranks_subsample.shape[1]) + ) + + # create lower triangular ones + template_mat = scipy.sparse.tril(np.ones(matches.shape), format="csr") + + # for every experiment, get C comparisons matrix + for _, row in pseudotime_ranks_subsample.iterrows(): + + # reorder template matrix to match real order + # basically, we want to move column #index to be column #argsort + # and same for rows + # where #index is an index into the lower-triangular matrix created above + # and #argsort is the argsort of the real order + + # e.g. suppose the values are [4,1,2,3] + # then the argsort is [2,3,4,1] + # the index (as always) into the lower-triangular matrix is [1,2,3,4] to start + # (in actuality the index and argsort would be zero-indexed. not the raw values) + # so you would move column 1 to be column 2, column 2 to be column 3, + # column 3 to be column 4, and column 4 to be column 1; and same for rows. + # i.e. column #index --> column #argsort + + # we achieve this by creating a series from the argsort, so we will have + # 1 2 + # 2 3 + # 3 4 + # 4 1 + + # then we sort the values + # that rearranges the index into 4,1,2,3 + # which is the order we put rows and columns of the lower-triangular matrix into. + + reorder_template = pd.Series(np.argsort(row.values)).sort_values().index.values + # note: this is a copy not a view unfortunately. + matches += template_mat[reorder_template, :][:, reorder_template] + + # maybe we can cut the below and just average? + + # # change 0s to -1s after the fact + # # x - y = 2x - (x+y) , where x is number of 1s, y is number of -1s + # # we have only been summing x, not x-y, so far + # matches *= 2 + # # Have to convert to dense here + # # subtracting a nonzero scalar from a sparse matrix is not supported + # matches = matches.toarray() - n_matches + + # # compute sample probabilities + # # Q = np.sum(matches, axis=0) + # Q = matches # already summed + # Q += 1 * n_matches + # Q *= 0.5 + # Q *= 1.0 / n_matches + Q = matches + + # normalize each row to have same euclidean norm + + Q_normalized = preprocessing.normalize(Q, norm="l2") + + Q_adata = anndata.AnnData(Q_normalized) + + # import scanpy as sc + sc.pp.neighbors(Q_adata, use_rep="X") + + # laplacian of the adjacency matrix of the kNN graph + laplacian_mat = laplacian(Q_adata.uns["neighbors"]["connectivities"], normed=False) + + _, eigenvecs = eigsh(laplacian_mat, 2, which="SM", tol=1e-8) + + # take second smallest eigenvalue + # order along it + labels = pseudotime_ranks_subsample.columns + order = [label for (_, label) in sorted(zip(eigenvecs[:, 1], labels))] + # assign orders 0 -> 1, normalize / num_cells + spectral_order_series = pd.Series( + np.arange(len(order)) / len(order), index=order, name="spectral_order" + ) + + # want no repeats in the order, i.e. all value counts should be 1 + if not all(spectral_order_series.reset_index()["index"].value_counts() == 1): + raise ValueError("Unexpected repeats in the spectral ordering") + + # sometimes eigenvector signs will be flipped, giving the reverse ordering. + # in these cases we should just reverse the ordering. + + # to determine whether this is the case: + # choose a cell we know to be closer to the start (or end), and ensure it is closer to the start (or end). + # for instance, run the mean ensembling, check whether the start or end point is ahead in the spectral ordering. + # flip order as needed to match. + + # another way to do this: + # look at mean spectral order of the root cells of all trajectories + # flip order if needed. + # (note: we subsampled the cells before running spectral ordering. + # so instead of using 0-position, use min-position cells from the trajectories.) + filtered_trajectories = trajectory_df[ + (trajectory_df[root_cell_key].isin(matches_sample)) + & (trajectory_df[barcode_key].isin(cells_sample)) + ] + if ( + spectral_order_series[ + filtered_trajectories.loc[ + filtered_trajectories.groupby("root_cell_key")["pseudotime"].idxmin() + ][barcode_key] + ].median() + > 0.5 + ): + spectral_order_series = 1 - spectral_order_series + + return spectral_order_series + + +def compare_trajectories( + trajectory_1, + trajectory_2, + trajectory_1_label="Trajectory 1", + trajectory_2_label="Trajectory 2", + title=None, +): + """[summary] + + # per-cell difference in ordering, hist + + # adata.obs["mean_trajectory"], adata.obs["spectral_order"] + # xlabel="Mean over percentile normalized trajectories" + # ylabel="Spectral order" + # title="Spectral vs mean pseudotime ensemble" + + :param trajectory_1: [description] + :type trajectory_1: [type] + :param trajectory_2: [description] + :type trajectory_2: [type] + :param trajectory_1_label: [description], defaults to 'Trajectory 1' + :type trajectory_1_label: str, optional + :param trajectory_2_label: [description], defaults to 'Trajectory 2' + :type trajectory_2_label: str, optional + :param title: [description], defaults to None + :type title: [type], optional + :return: [description] + :rtype: [type] + """ + + # merge indexes + compare_aggregates = pd.merge( + trajectory_1, trajectory_2, left_index=True, right_index=True, how="inner" + ) + trajectory_1, trajectory_2 = ( + compare_aggregates[trajectory_1.name], + compare_aggregates[trajectory_2.name], + ) + # print(compare_aggregates.shape) + + compare_aggregates["diff"] = trajectory_1 - trajectory_2 + compare_aggregates["abs_diff"] = compare_aggregates["diff"].abs() + # compare_aggregates.head() + + fig, ax = plt.subplots(figsize=(8, 8)) + plt.scatter(trajectory_1, trajectory_2, s=1, zorder=10) + + # plot y=x + plt.plot([0, 1], [0, 1], "k", alpha=0.75, zorder=0) + + ax.set_aspect("equal") + + plt.xlabel(trajectory_1_label) + plt.ylabel(trajectory_2_label) + if title is not None: + plt.title(title) + sns.despine() + + return fig, compare_aggregates["abs_diff"].median() + + +def plot_pseudotime_density( + adata, cluster_label_key, value_key, figsize=(6, 8.5), **kwargs +): + """[summary] + + # fig = plot_pseudotime_density(adata, suptitle="Pseudotime Coembed (mean trajectory)", cluster_label_key="cluster_label",value_key="mean_trajectory") + + :param adata: [description] + :type adata: [type] + :param cluster_label_key: [description] + :type cluster_label_key: [type] + :param value_key: [description] + :type value_key: [type] + :param figsize: [description], defaults to (6, 8.5) + :type figsize: tuple, optional + :return: [description] + :rtype: [type] + """ + ## First, remove without one-cell clusters because we can't draw a density for those + + clusters_to_keep = adata.obs[cluster_label_key].value_counts() + clusters_to_keep = clusters_to_keep[clusters_to_keep > 1].index.tolist() + adata_plt = adata[adata.obs[cluster_label_key].isin(clusters_to_keep)].copy() + # print("Removed 1-cell clusters") + # print(adata.shape, "-->", adata_plt.shape) + + ## Set a new ylevel that only contains the remaining clusters + + # problem with using cluster_label_key as ylevel: + # it's possibly a Categorical so it still contains the empty levels, so now those will have empty rows attached + # just convert back to strings from categorical + adata_plt.obs[cluster_label_key] = adata_plt.obs[cluster_label_key].astype("str") + + ## Set row order + # sort rows by median value + ylevel_order = ( + adata_plt.obs.groupby(cluster_label_key)[value_key] + .median() + .sort_values() + .index.tolist() + ) + + return plots.stacked_density_plot( + adata_plt.obs, + row_var=cluster_label_key, + hue_var=cluster_label_key, + value_var=value_key, + figsize=figsize, + row_order=ylevel_order, + xlabel="Pseudotime", + **kwargs + ) diff --git a/tests/test_trajectory.py b/tests/test_trajectory.py new file mode 100644 index 0000000..5fc439f --- /dev/null +++ b/tests/test_trajectory.py @@ -0,0 +1,77 @@ +#!/usr/bin/env python + +"""Tests for `genetools` package.""" + +import pytest +import numpy as np +import pandas as pd +import random +import matplotlib + +matplotlib.use("Agg") + +from genetools import plots, trajectory + +random_seed = 12345 +np.random.seed(random_seed) +random.seed(random_seed) + + +""" +TODO: +choose roots + + # afterwards: + # adata[roots].obs["cluster_label"].value_counts() + # plot_umap_highlight_cells + +=== + +- Plot root or end points. with umap_scatter + +=== +run + + # col_names, pseudotime_vals, many_pt = run_pseudotime(adata, roots, ["full_barcode", "cluster_label"]) + + +=== + +get end points + + # then look at adata[roots].obs["cluster_label"].value_counts() + # but first need to ['full_barcode'] it + +=== + +stochasticity plot + +=== + +mean order +spectral order: + n_trajectories_sample = 100 # number of trajectories to look at + n_cells_sample = 4400 # number of cells to look at + root_cell_key = 'root_cell' + barcode_key = 'full_barcode' + After running: left merge into the obs. +compare_trajectories + + + +# def plot_trajectory(): +# fig, ax = genetools.plots.umap_scatter( +# adata.obs, +# hue_key="mean_trajectory", +# continuous_hue=True, +# label_key="cluster_label", +# marker_size=0.1, +# umap_1_key="coembed_umap_1", +# umap_2_key="coembed_umap_2", +# figsize=(8, 8), +# ) +# ax.set_title("Mean pseudotime on Coembed") +# savefig(fig, "out/scanpy/coembed.pseudotime.16wk.rob_coembed.umap.png", dpi=300) + + +""" From 9b87b5e16858857be939da55fb8ec42de8b974e8 Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Fri, 10 Jul 2020 12:36:53 -0700 Subject: [PATCH 02/10] Remove legacy trajectory api --- genetools/trajectory.py | 65 +++++++---------------------------------- 1 file changed, 11 insertions(+), 54 deletions(-) diff --git a/genetools/trajectory.py b/genetools/trajectory.py index e60dc14..55b4a5b 100644 --- a/genetools/trajectory.py +++ b/genetools/trajectory.py @@ -142,10 +142,11 @@ def stochasticity( # change bin ID to be left-offset of bin instead df["bin_id"] = df["bin_id"].apply(lambda interval: interval.left) - # Histogram: number of values in each bin - # for this analysis, we want this to be roughly uniform. - # that's why we use percentile normalized pseudotime values above! otherwise it won't be uniform. + # Note: + # expects percentile normalized pseudotime values, + # so that we have a roughly uniform number of values in each bin # df["bin_id"].hist() + # TODO: confirm percentile normalized input # group by bin ID, count unique barcodes in that bin cells_by_bin = df.groupby("bin_id", sort=False)["full_barcode"].nunique() @@ -164,6 +165,8 @@ def stochasticity( def mean_order(trajectory_df, barcode_key): """[summary] ## Ensemble: Mean trajectory + # Take each cell's mean across all trajectories + # "project cells onto this ensemble medoid trajectory" trajectories have different linear scales: @@ -190,7 +193,6 @@ def mean_order(trajectory_df, barcode_key): raise ValueError("Some cells are unreachable (NaN pseudotimes)") # Take each cell's mean across all trajectories - # "project cells onto this ensemble medoid trajectory" mean_trajectory = trajectory_df.groupby([barcode_key], sort=False)[ "pseudotime" ].mean() @@ -213,7 +215,6 @@ def spectral_order( n_cells_sample=None, ): """[summary] - Expect percent normalized input. Use a kNN graph from scanpy instead of computing a dense similarity matrix: @@ -254,29 +255,6 @@ def spectral_order( import anndata import scanpy as sc - # Legacy code from when we passed around arrays of individual trajectories - # (We might bring that API back): - - # pseudotime_ranks = pd.concat(pseudotime_vals, axis=1) - # pseudotime_ranks = pseudotime_ranks.apply(percentile_normalize, axis=0) - # pseudotime_ranks = pseudotime_ranks.T # n_experiments x n_cells, percentile normalized - - # print(pseudotime_ranks.shape) - # pseudotime_ranks - - # cells_sample = np.random.choice( - # pseudotime_ranks.columns, size=n_cells_sample, replace=False - # ) - # cells_sample[:5] - - # matches_sample = np.random.choice( - # pseudotime_ranks.index, size=n_trajectories_sample, replace=False - # ) - # matches_sample[:5] - - # pseudotime_ranks_subsample = pseudotime_ranks.loc[matches_sample][cells_sample] - # pseudotime_ranks_subsample.shape, pseudotime_ranks.shape - if not n_cells_sample: # default to 20% of cells n_cells_sample = int(trajectory_df[barcode_key].nunique() * 0.2) @@ -297,11 +275,6 @@ def spectral_order( ].pivot(index=root_cell_key, columns=barcode_key, values="pseudotime") # make P, the transition matrix - # # number of matches, as in a tournament where each match = all players compete against each other - # n_matches = pseudotime_ranks_subsample.shape[0] - # # number of players in the tournament - # n_cells = pseudotime_ranks_subsample.shape[1] - matches = scipy.sparse.csr_matrix( (pseudotime_ranks_subsample.shape[1], pseudotime_ranks_subsample.shape[1]) ) @@ -340,42 +313,26 @@ def spectral_order( # note: this is a copy not a view unfortunately. matches += template_mat[reorder_template, :][:, reorder_template] - # maybe we can cut the below and just average? - - # # change 0s to -1s after the fact - # # x - y = 2x - (x+y) , where x is number of 1s, y is number of -1s - # # we have only been summing x, not x-y, so far - # matches *= 2 - # # Have to convert to dense here - # # subtracting a nonzero scalar from a sparse matrix is not supported - # matches = matches.toarray() - n_matches - - # # compute sample probabilities - # # Q = np.sum(matches, axis=0) - # Q = matches # already summed - # Q += 1 * n_matches - # Q *= 0.5 - # Q *= 1.0 / n_matches + # compute sample probabilities Q = matches # normalize each row to have same euclidean norm - Q_normalized = preprocessing.normalize(Q, norm="l2") + # kNN graph Q_adata = anndata.AnnData(Q_normalized) - - # import scanpy as sc sc.pp.neighbors(Q_adata, use_rep="X") # laplacian of the adjacency matrix of the kNN graph laplacian_mat = laplacian(Q_adata.uns["neighbors"]["connectivities"], normed=False) + # take second smallest eigenvalue _, eigenvecs = eigsh(laplacian_mat, 2, which="SM", tol=1e-8) - # take second smallest eigenvalue - # order along it + # order along second smallest eigenvalue labels = pseudotime_ranks_subsample.columns order = [label for (_, label) in sorted(zip(eigenvecs[:, 1], labels))] + # assign orders 0 -> 1, normalize / num_cells spectral_order_series = pd.Series( np.arange(len(order)) / len(order), index=order, name="spectral_order" From 7f76aa987e0f7e8efd6a34a606b7e032129b16f5 Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Sat, 8 Jan 2022 23:58:25 -0800 Subject: [PATCH 03/10] More trajectory work and start skeleton of test interface --- genetools/helpers.py | 22 +++++ genetools/plots.py | 4 +- genetools/trajectory.py | 94 +++++++-------------- tests/baseline/test_plot_roots.png | Bin 0 -> 184989 bytes tests/baseline/test_stochasticity_plot.png | Bin 0 -> 33533 bytes tests/test_trajectory.py | 89 +++++++++++++++---- 6 files changed, 124 insertions(+), 85 deletions(-) create mode 100644 tests/baseline/test_plot_roots.png create mode 100644 tests/baseline/test_stochasticity_plot.png diff --git a/genetools/helpers.py b/genetools/helpers.py index fe0675d..9c9c219 100644 --- a/genetools/helpers.py +++ b/genetools/helpers.py @@ -60,6 +60,28 @@ def merge_into_left(left, right, **kwargs): return df +def sample_cells_from_clusters(obs_df, n_cells, cluster_key, cluster_names): + """[summary] + + :param obs_df: [description] + :type obs_df: [type] + :param n_cells: [description] + :type n_cells: [type] + :param cluster_key: [description] + :type cluster_key: [type] + :param cluster_names: [description] + :type cluster_names: [type] + :return: [description] + :rtype: [type] + """ + return ( + obs_df[obs_df[cluster_key].isin(cluster_names)] + .index.to_series() + .sample(n_cells) + .values + ) + + def horizontal_concat(df_left, df_right): """Concatenate df_right horizontally to df_left, with no checks for whether the indexes match, but confirming final shape. diff --git a/genetools/plots.py b/genetools/plots.py index 1f292f4..a9633da 100644 --- a/genetools/plots.py +++ b/genetools/plots.py @@ -143,8 +143,8 @@ def umap_scatter( # overlay: highlight cells if highlight_cell_names is not None: plt.scatter( - data.iloc[highlight_cell_names][umap_1_key], - data.iloc[highlight_cell_names][umap_2_key], + data.loc[highlight_cell_names][umap_1_key], + data.loc[highlight_cell_names][umap_2_key], s=highlight_marker_size, c=highlight_marker_color, zorder=highlight_zorder, diff --git a/genetools/trajectory.py b/genetools/trajectory.py index 55b4a5b..f21c2b5 100644 --- a/genetools/trajectory.py +++ b/genetools/trajectory.py @@ -6,111 +6,76 @@ import seaborn as sns -def monte_carlo_pseudotime(adata_input, roots, id_vars): +def monte_carlo_pseudotime(adata_input, root_cells): import scanpy as sc """[summary] + TODO: document what must be done to adata before running dpt. i.e. PCA, neighbors, and diffmap. + :param adata_input: [description] :type adata_input: [type] - :param roots: [description] - :type roots: [type] + :param root_cells: [description] + :type root_cells: [type] :param id_vars: Identifier variables in anndata obs to uniquely identify a cell. Will be passed through to output. :type id_vars: [type] :return: [description] :rtype: [type] """ - # TODO: can we avoid passing in id_vars, and just set 'full_barcode' from obs_names adata = adata_input.copy() - if "root_cell" in id_vars or "pseudotime" in id_vars: - raise ValueError("Cannot include 'root_cell' or 'pseudotime' in id_vars") - - # root cell names - col_names = [] - - # each entry is one trajectory from one root cell: a Series, values are the pseudotime values per-cell, and name is the root cell + # each entry is one trajectory from one root cell: + # # a Series, values are the pseudotime values per-cell, and name is the root cell pseudotime_vals = [] - for root_cell in roots: - # Set root cell + for root_cell in root_cells: + # Set root cell ID adata.uns["iroot"] = np.where(adata.obs.index == root_cell)[0][0] - # Run DPT. Scanpy stores output in obs 'dpt_pseudotime + # Run DPT. Scanpy stores output in obs 'dpt_pseudotime' sc.tl.dpt(adata, n_dcs=min(adata.obsm["X_diffmap"].shape[1], 10), copy=False) # Save this trajectory - output = adata.obs["dpt_pseudotime"].copy() - output.name = root_cell - col_names.append(output.name) - pseudotime_vals.append(output) + pseudotime_vals.append(adata.obs["dpt_pseudotime"].rename(root_cell)) # percentile normalize each pseudotime trajectory pseudotime_vals = pd.concat(pseudotime_vals, axis=1) pseudotime_vals_percentile_normalized = pseudotime_vals.apply( stats.percentile_normalize, axis=0 ) - if pseudotime_vals_percentile_normalized.shape != pseudotime_vals.shape: - raise ValueError() # Create dataframe where each row is a cell's value in a trajectory from a specific root cell. # i.e. convert to format: one value per cell, times # experiments - df = pd.concat([adata_input.obs] + pseudotime_vals_percentile_normalized, axis=1) df = pd.melt( - df.reset_index(), - id_vars=id_vars, - value_vars=col_names, + pseudotime_vals_percentile_normalized.rename_axis("cell_barcode").reset_index(), + id_vars="cell_barcode", + value_vars=root_cells, var_name="root_cell", value_name="pseudotime", ) - if df.shape[0] != adata_input.obs.shape[0] * len(roots): - raise ValueError( - "Should have one value per cell, times the number of experiments" - ) - return df -def choose_roots(adata, n_roots, cluster_key, cluster_names): - """[summary] - - - - :param n_roots: [description] - :type n_roots: [type] - :param cluster_key: [description] - :type cluster_key: [type] - :param cluster_names: [description] - :type cluster_names: [type] - :return: [description] - :rtype: [type] - """ - return ( - adata.obs[adata.obs[cluster_key].isin(cluster_names)] - .index.to_series() - .sample(n_roots) - .values - ) - - -def get_end_points(trajectory_df, id_vars): - """[summary] +def get_cells_at_percentile(trajectory_df, percentile): + """get points at percentile along trajectories :param trajectory_df: [description] :type trajectory_df: [type] - :param id_vars: [description] - :type id_vars: [type] + :param percentile: [description] (should be between 0 and 1) + :type percentile: [type] :return: [description] :rtype: [type] """ - return trajectory_df.loc[ - trajectory_df.groupby("root_cell", sort=False)["pseudotime"].idxmax() - ][id_vars] + if not (0.0 <= percentile <= 1.0): + raise ValueError("percentile must be between 0 and 1") + df = trajectory_df.copy() + df["diff_pseudotime"] = (df["pseudotime"] - percentile).abs() + return df.loc[df.groupby("root_cell", sort=False)["diff_pseudotime"].idxmin()] -def stochasticity( +def plot_stochasticity( trajectory_df, xlabel="Pseudotime", ylabel="Number of unique barcodes in bin", @@ -149,10 +114,11 @@ def stochasticity( # TODO: confirm percentile normalized input # group by bin ID, count unique barcodes in that bin - cells_by_bin = df.groupby("bin_id", sort=False)["full_barcode"].nunique() + cells_by_bin = df.groupby("bin_id", sort=False)["cell_barcode"].nunique() fig = plt.figure() plt.scatter(cells_by_bin.index, cells_by_bin.values) + plt.xlim(0, 1) if xlabel is not None: plt.xlabel(xlabel) if ylabel is not None: @@ -162,7 +128,7 @@ def stochasticity( return fig -def mean_order(trajectory_df, barcode_key): +def mean_order(trajectory_df, barcode_key="cell_barcode"): """[summary] ## Ensemble: Mean trajectory # Take each cell's mean across all trajectories @@ -210,7 +176,7 @@ def mean_order(trajectory_df, barcode_key): def spectral_order( trajectory_df, root_cell_key, - barcode_key, + barcode_key="cell_barcode", n_trajectories_sample=None, n_cells_sample=None, ): @@ -372,7 +338,7 @@ def spectral_order( return spectral_order_series -def compare_trajectories( +def compare_orders( trajectory_1, trajectory_2, trajectory_1_label="Trajectory 1", @@ -438,8 +404,6 @@ def plot_pseudotime_density( ): """[summary] - # fig = plot_pseudotime_density(adata, suptitle="Pseudotime Coembed (mean trajectory)", cluster_label_key="cluster_label",value_key="mean_trajectory") - :param adata: [description] :type adata: [type] :param cluster_label_key: [description] diff --git a/tests/baseline/test_plot_roots.png b/tests/baseline/test_plot_roots.png new file mode 100644 index 0000000000000000000000000000000000000000..ddc290e7907f2129ea831bd370f64d01b27097b2 GIT binary patch literal 184989 zcmcG$2T)UM*fknJigZMJ6A%?as`MsBzyb&&y(1vK*U+RZO%&-xq$*vcmxPW|f+D>H zqy(fBAV4Vp8$I9u&)k_icjn%?8JV1L!rs~Yead>)T5n!G(Nv=#WhR9{AQTVORdgT_ zf&~Z!e~^R-{N~;V3l8vf$y4cpE(v%9l01(AuP?i)8+t+@RF=3Oyb^_Cd+>OsOy)tG3y(sYxD797m=RvW>{%8f{P~^awX!vqVz2;ooDw;aVsMB-&>QZ|Ia~nR+Nt>YwXfx{Y4~@ zfuh2~!i-VvGTSZhw#5WeV^;<|%iZuArqh9~};W$?IEs0xA(rq3h*mh*qx7VAB?4d@}6#`BZ)N6k|IU4NsAnB2m43z~Ae&e^&>jIeO_ z>VK#GQ1O~Ncx8a;hGtfub)uZAiHXU}3b~MT*=FDEBKwh|WIp|hG^1O%qwMc!IX+~QPtQLu=UPc3kB={(U_A<32Uya78u z;fto?AetZ1QW#`*k$0Jqyh2Osd0>T&F06m?gG>M3|Aa6F?k~ihnTaV``vS6)lT~zd zu0TLo^ogY!{}<5wOg8cPo6}R43rV`j>81KHIe(>OdQJRLz 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+# TODO: multiple root clusters +root_cluster = "B" -""" -TODO: -choose roots - # afterwards: - # adata[roots].obs["cluster_label"].value_counts() - # plot_umap_highlight_cells +@pytest.fixture(scope="module") +def roots(adata): + # Choose roots + return helpers.sample_cells_from_clusters( + adata.obs, n_roots, "louvain", [root_cluster] + ) -=== -- Plot root or end points. with umap_scatter +def test_roots(adata, roots): + # Confirm roots come from the right clusters + assert len(roots) == n_roots + assert all(adata[roots].obs["louvain"] == root_cluster) -=== -run - # col_names, pseudotime_vals, many_pt = run_pseudotime(adata, roots, ["full_barcode", "cluster_label"]) +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}) +def test_plot_roots(adata, roots): + # Plot root points over umap + fig, _ = plots.umap_scatter( + data=adata.obs, + umap_1_key="umap_1", + umap_2_key="umap_2", + hue_key="louvain", + label_key="louvain", + highlight_cell_names=roots, + ) + return fig -=== +@pytest.fixture(scope="module") +def trajectories(adata, roots): + # run pseudotime + return trajectory.monte_carlo_pseudotime(adata, roots) -get end points - # then look at adata[roots].obs["cluster_label"].value_counts() - # but first need to ['full_barcode'] it +def test_trajectory_shape(trajectories, adata, roots): + # Should have one value per cell, times the number of experiments + assert trajectories.shape[0] == adata.obs.shape[0] * len(roots) -=== -stochasticity plot +def test_trajectory_ranges(trajectories): + assert all(trajectories["pseudotime"] >= 0.0) + assert all(trajectories["pseudotime"] <= 1.0) + assert not any(trajectories["pseudotime"].isna()) + + +def test_trajectory_colnames(trajectories): + expected_cols = ["cell_barcode", "root_cell", "pseudotime"] + for col in expected_cols: + assert col in trajectories.columns + + +def test_get_end_points(trajectories, roots): + df = trajectory.get_cells_at_percentile(trajectories, 1.0) + assert df.shape[0] == len(roots) + assert all(df["pseudotime"]) == 1.0 + + +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}) +def test_stochasticity_plot(trajectories): + return trajectory.plot_stochasticity(trajectories) + + +""" +Tutorial order: +- roots choose, plot, get value counts by cluster +- run trajectories +- end points get value counts by cluster, and plot +- stochasticity plot + + +TODO: === @@ -58,6 +105,8 @@ compare_trajectories +left merge mean_trajectory back + # def plot_trajectory(): # fig, ax = genetools.plots.umap_scatter( @@ -74,4 +123,8 @@ # savefig(fig, "out/scanpy/coembed.pseudotime.16wk.rob_coembed.umap.png", dpi=300) +# plot density +# fig = plot_pseudotime_density(adata, suptitle="Pseudotime Coembed (mean trajectory)", cluster_label_key="cluster_label",value_key="mean_trajectory") + + """ From a8ac47c1be6a49e132ba7c7c4a2d8cac0a146094 Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Wed, 12 Aug 2020 21:42:34 -0700 Subject: [PATCH 04/10] Test mean and spectral orderings --- genetools/trajectory.py | 11 ++- .../test_mean_vs_spectral_orderings.png | Bin 0 -> 32830 bytes .../baseline/test_plot_trajectory_on_umap.png | Bin 0 -> 132641 bytes tests/test_trajectory.py | 91 ++++++++++++------ 4 files changed, 69 insertions(+), 33 deletions(-) create mode 100644 tests/baseline/test_mean_vs_spectral_orderings.png create mode 100644 tests/baseline/test_plot_trajectory_on_umap.png diff --git a/genetools/trajectory.py b/genetools/trajectory.py index f21c2b5..6439e6b 100644 --- a/genetools/trajectory.py +++ b/genetools/trajectory.py @@ -162,12 +162,12 @@ def mean_order(trajectory_df, barcode_key="cell_barcode"): mean_trajectory = trajectory_df.groupby([barcode_key], sort=False)[ "pseudotime" ].mean() - if any(mean_trajectory.isna()): - raise ValueError() # renormalize mean_trajectory = pd.Series( - stats.percentile_normalize(mean_trajectory), index=mean_trajectory.index + stats.percentile_normalize(mean_trajectory), + index=mean_trajectory.index, + name="mean_order", ) return mean_trajectory @@ -175,7 +175,7 @@ def mean_order(trajectory_df, barcode_key="cell_barcode"): def spectral_order( trajectory_df, - root_cell_key, + root_cell_key="root_cell", barcode_key="cell_barcode", n_trajectories_sample=None, n_cells_sample=None, @@ -306,6 +306,7 @@ def spectral_order( # want no repeats in the order, i.e. all value counts should be 1 if not all(spectral_order_series.reset_index()["index"].value_counts() == 1): + # TODO: try not any(spectral_order_series.index.duplicated()) for clarity instead? raise ValueError("Unexpected repeats in the spectral ordering") # sometimes eigenvector signs will be flipped, giving the reverse ordering. @@ -328,7 +329,7 @@ def spectral_order( if ( spectral_order_series[ filtered_trajectories.loc[ - filtered_trajectories.groupby("root_cell_key")["pseudotime"].idxmin() + filtered_trajectories.groupby(root_cell_key)["pseudotime"].idxmin() ][barcode_key] ].median() > 0.5 diff --git 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as pd @@ -83,45 +84,79 @@ def test_stochasticity_plot(trajectories): return trajectory.plot_stochasticity(trajectories) +@pytest.fixture(scope="module") +def mean_ordering(trajectories): + return trajectory.mean_order(trajectories, "cell_barcode") + + +@pytest.fixture(scope="module") +def spectral_ordering(trajectories): + return trajectory.spectral_order(trajectories, "root_cell", "cell_barcode") + + +@pytest.mark.mpl_image_compare +def test_mean_vs_spectral_orderings( + adata, trajectories, mean_ordering, spectral_ordering +): + # confirm basics about both series (use ", series.name" to emit series name in errors) + for series in [mean_ordering, spectral_ordering]: + # check range + assert all(series >= 0.0), series.name + assert all(series <= 1.0), series.name + # no NaNs + assert not any(series.isna()), series.name + # every cell barcode included (exception: spectral order takes a subset) + assert series.shape[0] <= trajectories["cell_barcode"].nunique(), series.name + # no duplicate barcodes, i.e. only one entry per cell barcode + assert not any(series.index.duplicated()), series.name + # test merge back into adata obs, to confirm mergeability and validate 1:1 + helpers.merge_into_left(adata.obs, series) + + # compare orderings + fig, median_abs_diff = trajectory.compare_orders( + mean_ordering, + spectral_ordering, + trajectory_1_label="Mean order", + trajectory_2_label="Spectral order", + title="Mean vs spectral", + ) + assert median_abs_diff <= 0.2 + return fig + + +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}) +def test_plot_trajectory_on_umap(adata, mean_ordering): + plot_data = helpers.merge_into_left(adata.obs, mean_ordering) + fig, ax = plots.umap_scatter( + data=plot_data, + umap_1_key="umap_1", + umap_2_key="umap_2", + hue_key="mean_order", + continuous_hue=True, + label_key="louvain", + ) + ax.set_title("Mean pseudotime trajectory") + return fig + + """ Tutorial order: - roots choose, plot, get value counts by cluster - run trajectories - end points get value counts by cluster, and plot - stochasticity plot - - -TODO: - -=== - -mean order -spectral order: +- mean order +- spectral order: n_trajectories_sample = 100 # number of trajectories to look at n_cells_sample = 4400 # number of cells to look at root_cell_key = 'root_cell' barcode_key = 'full_barcode' - After running: left merge into the obs. -compare_trajectories - - -left merge mean_trajectory back - - -# def plot_trajectory(): -# fig, ax = genetools.plots.umap_scatter( -# adata.obs, -# hue_key="mean_trajectory", -# continuous_hue=True, -# label_key="cluster_label", -# marker_size=0.1, -# umap_1_key="coembed_umap_1", -# umap_2_key="coembed_umap_2", -# figsize=(8, 8), -# ) -# ax.set_title("Mean pseudotime on Coembed") -# savefig(fig, "out/scanpy/coembed.pseudotime.16wk.rob_coembed.umap.png", dpi=300) + (After running: left merge into the obs?) +- compare_trajectories +- left merge mean_trajectory back +- plot trajectory on umap +TODO: # plot density # fig = plot_pseudotime_density(adata, suptitle="Pseudotime Coembed (mean trajectory)", cluster_label_key="cluster_label",value_key="mean_trajectory") From 7b586aeae2ac44f866cb02759892293f5a9af6dd Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Sun, 9 Jan 2022 00:00:46 -0800 Subject: [PATCH 05/10] Stacked density plot --- genetools/plots.py | 112 +++++++++++++++--- genetools/trajectory.py | 52 -------- tests/baseline/test_stacked_density_plot.png | Bin 0 -> 48152 bytes ...stacked_density_plot_overlap_no_labels.png | Bin 0 -> 34435 bytes tests/test_trajectory.py | 33 +++++- 5 files changed, 121 insertions(+), 76 deletions(-) create mode 100644 tests/baseline/test_stacked_density_plot.png create mode 100644 tests/baseline/test_stacked_density_plot_overlap_no_labels.png diff --git a/genetools/plots.py b/genetools/plots.py index a9633da..039a037 100644 --- a/genetools/plots.py +++ b/genetools/plots.py @@ -287,11 +287,89 @@ def horizontal_stacked_bar_plot( def stacked_density_plot( + data, + cluster_label_key, + value_key, + xlabel=None, + suptitle=None, + figsize=(6, 8.5), + palette=None, + overlap=False, + **kwargs +): + """[summary] + Plot probability densities. value_key should range from 0 to 1. + + :param data: [description] + :type data: [type] + :param cluster_label_key: [description] + :type cluster_label_key: [type] + :param value_key: [description] + :type value_key: [type] + :param xlabel: [description], defaults to None + :type xlabel: [type], optional + :param suptitle: [description], defaults to None + :type suptitle: [type], optional + :param figsize: [description], defaults to (6, 8.5) + :type figsize: tuple, optional + :param palette: [description], defaults to None + :type palette: [type], optional + :param overlap: [description], defaults to False + :type overlap: bool, optional + :return: [description] + :rtype: [type] + """ + ## 1. remove without one-cell clusters because we can't draw a density for those + + clusters_to_keep = data[cluster_label_key].value_counts() + clusters_to_keep = clusters_to_keep[clusters_to_keep > 1].index.tolist() + plot_df = data[data[cluster_label_key].isin(clusters_to_keep)].copy() + + # TODO: bring back debug logging: + # print("Removed 1-cell clusters") + # print(data.shape, "-->", plot_df.shape) + + ## 2. Set a new ylevel that only contains the remaining clusters + + # problem with using cluster_label_key as ylevel: + # it's possibly a Categorical so it still contains the empty levels, so now those will have empty rows attached + # just convert back to strings from categorical + plot_df[cluster_label_key] = plot_df[cluster_label_key].astype("str") + + ## 3. Set row order + + # sort rows by median value + # so we have nicely increasing densities as we go down the plot + ylevel_order = ( + plot_df.groupby(cluster_label_key)[value_key] + .median() + .sort_values() + .index.tolist() + ) + + ## 4. Plot. + + return _stacked_density_facetgrid( + plot_df, + row_var=cluster_label_key, + hue_var=cluster_label_key, + value_var=value_key, + xlabel=xlabel, + overlap=overlap, + suptitle=suptitle, + figsize=figsize, + row_order=ylevel_order, + palette=palette, + **kwargs + ) + + +def _stacked_density_facetgrid( data, row_var, hue_var, value_var, - xlabel, + xlabel=None, col_var=None, overlap=False, suptitle=None, @@ -301,10 +379,13 @@ def stacked_density_plot( palette=None, ): """ - Multiple density plot. + Customizable multiple rows+columns density plot. + Adapted from old work at https://github.com/hammerlab/infino/blob/develop/analyze_cut.py#L912 For row_order, consider row_order=reversed(list(range(data.ylevel.values.max()+1))) + + May want to do this wrapped in `with sns.plotting_context("notebook"):` ? """ with sns.plotting_context("notebook"): @@ -333,25 +414,26 @@ def stacked_density_plot( def label(**kwargs): """ - kwargs is e.g.: {'color': (0.4918017777777778, 0.25275644444444445, 0.3333333333333333), 'label': 'Name of the row'} + default kwargs are color (RGB tuple) and label (name of the row) + other text options to consider passing in as kwargs: + - fontweight="bold" + - ha="right" + - bbox=dict(facecolor='yellow', alpha=0.3) """ - color = kwargs["color"] - label = kwargs["label"] + color = kwargs.pop("color") + label = kwargs.pop("label") ax = plt.gca() # map() changes current axis repeatedly # x=1 if plot_on_right else 0; ha="right" if plot_on_right else "left", ax.text( 1.25, 0.5, label, - # fontweight="bold", color=color, - # ha="right", ha="left", va="center", transform=ax.transAxes, fontsize="x-small", - # fontsize='x-large', #15, - # bbox=dict(facecolor='yellow', alpha=0.3) + **kwargs ) g.map(label) @@ -372,23 +454,17 @@ def label(**kwargs): g.despine(bottom=True, left=True) # fix x axis - g.set_xlabels(xlabel) + if xlabel is not None: + g.set_xlabels(xlabel) # resize if figsize: g.fig.set_size_inches(figsize[0], figsize[1]) - else: - cur_size = g.fig.get_size_inches() - increase_vertical = 3 # 7 #4 # 3 - g.fig.set_size_inches(cur_size[0], cur_size[1] + increase_vertical) if suptitle is not None: g.fig.suptitle(suptitle, fontsize="medium") - # tighten - g.fig.tight_layout() - - # then reoverlap + # overlap if overlap: g.fig.subplots_adjust(hspace=-0.1) diff --git a/genetools/trajectory.py b/genetools/trajectory.py index 6439e6b..a9f45b4 100644 --- a/genetools/trajectory.py +++ b/genetools/trajectory.py @@ -398,55 +398,3 @@ def compare_orders( sns.despine() return fig, compare_aggregates["abs_diff"].median() - - -def plot_pseudotime_density( - adata, cluster_label_key, value_key, figsize=(6, 8.5), **kwargs -): - """[summary] - - :param adata: [description] - :type adata: [type] - :param cluster_label_key: [description] - :type cluster_label_key: [type] - :param value_key: [description] - :type value_key: [type] - :param figsize: [description], defaults to (6, 8.5) - :type figsize: tuple, optional - :return: [description] - :rtype: [type] - """ - ## First, remove without one-cell clusters because we can't draw a density for those - - clusters_to_keep = adata.obs[cluster_label_key].value_counts() - clusters_to_keep = clusters_to_keep[clusters_to_keep > 1].index.tolist() - adata_plt = adata[adata.obs[cluster_label_key].isin(clusters_to_keep)].copy() - # print("Removed 1-cell clusters") - # print(adata.shape, "-->", adata_plt.shape) - - ## Set a new ylevel that only contains the remaining clusters - - # problem with using cluster_label_key as ylevel: - # it's possibly a Categorical so it still contains the empty levels, so now those will have empty rows attached - # just convert back to strings from categorical - adata_plt.obs[cluster_label_key] = adata_plt.obs[cluster_label_key].astype("str") - - ## Set row order - # sort rows by median value - ylevel_order = ( - adata_plt.obs.groupby(cluster_label_key)[value_key] - .median() - .sort_values() - .index.tolist() - ) - - return plots.stacked_density_plot( - adata_plt.obs, - row_var=cluster_label_key, - hue_var=cluster_label_key, - value_var=value_key, - figsize=figsize, - row_order=ylevel_order, - xlabel="Pseudotime", - **kwargs - ) diff --git a/tests/baseline/test_stacked_density_plot.png b/tests/baseline/test_stacked_density_plot.png new file mode 100644 index 0000000000000000000000000000000000000000..e21d2e3a1dc6834ef8063c5003e1d563cd979330 GIT binary patch literal 48152 zcmeFZWn9!>^glR5cb9a^Agw6fDM+J~v`BaN&>^V^h#)0`(jnccpmaCV-3-m%`Tkz} z|L;E72fL4VUJA^7Zk&70x$pBn=U#}`P*cRmp~itgAo$Of(c0bJS%jDOe?RAOa<$^kI~JLNKo}v<<)n4IGj`{F zGRV#{CGRE+el+;`?ms{aU0M>(b5#)|`skhAZYuVy=%Lhczw#WdZM>7`(;WqiO6tk9 z@i_K|^%3vg9#TS`Lj_0TL={J>G&8u_+pE%*lgnGpdGXOd?Rcl|s)J2m5{Md#8jBXX z;V0>X8`=jX${gs-;MMow?+!{7cwtau@PYTav=$H+C2=g 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a/tests/test_trajectory.py +++ b/tests/test_trajectory.py @@ -8,6 +8,7 @@ import pandas as pd import random import matplotlib +import seaborn as sns matplotlib.use("Agg") @@ -139,6 +140,31 @@ def test_plot_trajectory_on_umap(adata, mean_ordering): return fig +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}) +def test_stacked_density_plot(adata, mean_ordering): + plot_data = helpers.merge_into_left(adata.obs, mean_ordering) + return plots.stacked_density_plot( + data=plot_data, + cluster_label_key="louvain", + value_key="mean_order", + xlabel="Pseudotime", + suptitle="Mean pseudotime trajectory", + ) + + +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}, backend="Agg") +def test_stacked_density_plot_overlap_no_labels(adata, mean_ordering): + plot_data = helpers.merge_into_left(adata.obs, mean_ordering) + return plots.stacked_density_plot( + data=plot_data, + cluster_label_key="louvain", + value_key="mean_order", + figsize=(6, 6), + palette=sns.color_palette("Set2"), + overlap=True, + ) + + """ Tutorial order: - roots choose, plot, get value counts by cluster @@ -155,11 +181,6 @@ def test_plot_trajectory_on_umap(adata, mean_ordering): - compare_trajectories - left merge mean_trajectory back - plot trajectory on umap - -TODO: - -# plot density -# fig = plot_pseudotime_density(adata, suptitle="Pseudotime Coembed (mean trajectory)", cluster_label_key="cluster_label",value_key="mean_trajectory") - +- plot stacked density """ From 0e560ab892d4dde335f44aa3665a1839590d63e2 Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Sun, 9 Jan 2022 00:03:51 -0800 Subject: [PATCH 06/10] fixup! Stacked density plot --- genetools/plots.py | 5 +++-- tests/baseline/test_stacked_density_plot.png | Bin 48152 -> 41872 bytes ...stacked_density_plot_overlap_no_labels.png | Bin 34435 -> 31139 bytes tests/test_trajectory.py | 2 +- 4 files changed, 4 insertions(+), 3 deletions(-) diff --git a/genetools/plots.py b/genetools/plots.py index 039a037..5a72b77 100644 --- a/genetools/plots.py +++ b/genetools/plots.py @@ -384,8 +384,6 @@ def _stacked_density_facetgrid( Adapted from old work at https://github.com/hammerlab/infino/blob/develop/analyze_cut.py#L912 For row_order, consider row_order=reversed(list(range(data.ylevel.values.max()+1))) - - May want to do this wrapped in `with sns.plotting_context("notebook"):` ? 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b/tests/test_trajectory.py @@ -152,7 +152,7 @@ def test_stacked_density_plot(adata, mean_ordering): ) -@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}, backend="Agg") +@pytest.mark.mpl_image_compare(savefig_kwargs={"bbox_inches": "tight"}) def test_stacked_density_plot_overlap_no_labels(adata, mean_ordering): plot_data = helpers.merge_into_left(adata.obs, mean_ordering) return plots.stacked_density_plot( From dee51179909b87e17ec6047d3c2e4561b4c6b167 Mon Sep 17 00:00:00 2001 From: Maxim Zaslavsky Date: Sun, 9 Jan 2022 00:05:04 -0800 Subject: [PATCH 07/10] Tests --- genetools/trajectory.py | 8 ++---- ...stacked_density_plot_overlap_no_labels.png | Bin 31139 -> 36596 bytes tests/test_trajectory.py | 26 +++++++++++++++++- 3 files changed, 28 insertions(+), 6 deletions(-) diff --git a/genetools/trajectory.py b/genetools/trajectory.py index a9f45b4..02eb6d0 100644 --- a/genetools/trajectory.py +++ b/genetools/trajectory.py @@ -221,6 +221,9 @@ def spectral_order( import anndata import scanpy as sc + if any(trajectory_df["pseudotime"].isna()): + raise ValueError("Some cells are unreachable (NaN pseudotimes)") + if not n_cells_sample: # default to 20% of cells n_cells_sample = int(trajectory_df[barcode_key].nunique() * 0.2) @@ -304,11 +307,6 @@ def spectral_order( np.arange(len(order)) / len(order), index=order, name="spectral_order" ) - # want no repeats in the order, i.e. all value counts should be 1 - if not all(spectral_order_series.reset_index()["index"].value_counts() == 1): - # TODO: try not any(spectral_order_series.index.duplicated()) for clarity instead? - raise ValueError("Unexpected repeats in the spectral ordering") - # sometimes eigenvector signs will be flipped, giving the reverse ordering. # in these cases we should just reverse the ordering. diff --git a/tests/baseline/test_stacked_density_plot_overlap_no_labels.png b/tests/baseline/test_stacked_density_plot_overlap_no_labels.png index 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