Open In Colab Open In nbviewer

Pancreatic endocrinogenesis

This tutorial uses raw data from scvelo package. Special thanks go to the scvelo team!

[ ]:

# get the latest version from pypi
# for other installations approaches, see https://dynamo-release.readthedocs.io/en/latest/ten_minutes_to_dynamo.html#how-to-install
!pip install dynamo-release --upgrade --quiet

[1]:

# from IPython.core.display import display, HTML
# display(HTML("<style>.container { width:90% !important; }</style>"))
# %matplotlib inline
import warnings
warnings.filterwarnings('ignore')

import dynamo as dyn

this is like R’s sessionInfo()

[2]:

dyn.get_all_dependencies_version()
package dynamo-release umap-learn anndata cvxopt hdbscan loompy matplotlib numba numpy pandas pynndescent python-igraph scikit-learn scipy seaborn setuptools statsmodels tqdm trimap numdifftools colorcet
version 0.95.2 0.4.6 0.7.4 1.2.3 0.8.26 3.0.6 3.3.0 0.51.0 1.19.1 1.1.1 0.4.8 0.8.2 0.23.2 1.5.2 0.9.0 49.6.0 0.11.1 4.48.2 1.0.12 0.9.39 2.0.2 
[3]:

# run dynamo to get RNA velocity

dyn.configuration.set_figure_params('dynamo', background='white')

adata = dyn.sample_data.pancreatic_endocrinogenesis()

dyn.pp.recipe_monocle(adata, n_top_genes=1000, fg_kwargs={'shared_count': 20})

dyn.tl.dynamics(adata, model='stochastic')

dyn.tl.reduceDimension(adata, n_pca_components=30)

dyn.tl.cell_velocities(adata, method='pearson', other_kernels_dict={'transform': 'sqrt'})
dyn.pl.streamline_plot(adata, color=['clusters'], basis='umap', show_legend='on data', show_arrowed_spines=True)


estimating gamma: 100%|██████████| 1000/1000 [00:27<00:00, 36.67it/s]
calculating transition matrix via pearson kernel with sqrt transform.: 100%|██████████| 3696/3696 [00:04<00:00, 799.84it/s]
projecting velocity vector to low dimensional embedding...: 100%|██████████| 3696/3696 [00:00<00:00, 4314.30it/s]

<Figure size 600x400 with 0 Axes>
_images/pancreatic_endocrinogenesis_5_2.png 
[4]:

dyn.pl.phase_portraits(adata, genes=adata.var_names[adata.var.use_for_dynamics][:4], figsize=(6, 4), color='clusters')
_images/pancreatic_endocrinogenesis_6_0.png 
[5]:

dyn.pl.umap(adata, color=['clusters', 'clusters_coarse', "S_score", "G2M_score"], ncols=4, alpha=0.1)
_images/pancreatic_endocrinogenesis_7_0.png 
[6]:

dyn.pl.streamline_plot(adata, color=['clusters'], basis='umap', show_legend='on data')


<Figure size 600x400 with 0 Axes>
_images/pancreatic_endocrinogenesis_8_1.png 
[7]:

dyn.pl.cell_wise_vectors(adata, color=['clusters'], basis='umap', show_legend='on data', quiver_length=6, quiver_size=6, figsize=(8, 6), show_arrowed_spines=False)


<Figure size 800x600 with 0 Axes>
_images/pancreatic_endocrinogenesis_9_1.png 
[8]:

# ok some exciting vector field analysis

# you can set `verbose = 1/2/3` to obtain different levels of running information of vector field reconstruction

dyn.vf.VectorField(adata, basis='umap', pot_curl_div=True) # , M=1000, MaxIter=1000


Constructing diffusion graph from reconstructed vector field: 3696it [00:58, 63.53it/s]
Calculating 2-D curl: 100%|██████████| 3696/3696 [00:00<00:00, 16788.15it/s]
Calculating divergence: 100%|██████████| 3696/3696 [00:00<00:00, 13627.22it/s]

[9]:

dyn.pl.topography(adata, color=['clusters'], basis='umap', background='white',
                  streamline_color='black', show_legend='on data', terms=("streamline"))


<Figure size 600x400 with 0 Axes>
_images/pancreatic_endocrinogenesis_11_1.png 
[10]:

dyn.pl.topography(adata, basis='umap', background='white', color=['ntr', 'clusters'], streamline_color='black', show_legend='on data')
_images/pancreatic_endocrinogenesis_12_0.png 
[11]:

dyn.tl.cell_velocities(adata, basis='pca')
dyn.vf.VectorField(adata, basis='pca')
dyn.vf.speed(adata)
dyn.vf.divergence(adata)
dyn.vf.acceleration(adata)
dyn.vf.curl(adata)

projecting velocity vector to low dimensional embedding...:  10%|█         | 378/3696 [00:00<00:00, 3769.11it/s]

Using existing pearson_transition_matrix found in .obsp.

projecting velocity vector to low dimensional embedding...: 100%|██████████| 3696/3696 [00:00<00:00, 4015.84it/s]
Calculating divergence: 100%|██████████| 3696/3696 [00:00<00:00, 7500.08it/s]
Calculating acceleration: 100%|██████████| 3696/3696 [00:00<00:00, 292963.20it/s]
Calculating 2-D curl: 100%|██████████| 3696/3696 [00:00<00:00, 17275.38it/s]

[12]:

adata

[12]:

AnnData object with n_obs × n_vars = 3696 × 27998
    obs: 'clusters_coarse', 'clusters', 'S_score', 'G2M_score', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'Size_Factor', 'initial_cell_size', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'ntr', 'cell_cycle_phase', 'umap_ddhodge_div', 'umap_ddhodge_potential', 'curl_umap', 'divergence_umap', 'speed_umap', 'divergence_pca', 'acceleration_umap'
    var: 'highly_variable_genes', 'pass_basic_filter', 'log_m', 'score', 'log_cv', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics', 'use_for_transition'
    uns: 'clusters_coarse_colors', 'clusters_colors', 'day_colors', 'neighbors', 'pca', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'grid_velocity_umap', 'VecFld_umap', 'VecFld', 'grid_velocity_pca', 'VecFld_pca'
    obsm: 'X_pca', 'X_umap', 'X', 'cell_cycle_scores', 'velocity_umap', 'velocity_umap_SparseVFC', 'X_umap_SparseVFC', 'velocity_pca', 'velocity_pca_SparseVFC', 'X_pca_SparseVFC', 'acceleration_umap'
    layers: 'spliced', 'unspliced', 'X_spliced', 'X_unspliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'distances', 'connectivities', 'moments_con', 'pearson_transition_matrix', 'umap_ddhodge'

this just shows how flexible dynamo’ plotting function can be.

[13]:

import matplotlib.pyplot as plt
fig1, f1_axes = plt.subplots(ncols=2, nrows=2, constrained_layout=True, figsize=(12, 8))
f1_axes
f1_axes[0, 0] = dyn.pl.cell_wise_vectors(adata, color='umap_ddhodge_potential', pointsize=0.1, alpha = 0.7, ax=f1_axes[0, 0], quiver_length=6, quiver_size=6, save_show_or_return='return')
f1_axes[0, 1] = dyn.pl.grid_vectors(adata, color='speed_umap', ax=f1_axes[0, 1], quiver_length=12, quiver_size=12, save_show_or_return='return')
f1_axes[1, 0] = dyn.pl.streamline_plot(adata, color='divergence_pca', ax=f1_axes[1, 0], save_show_or_return='return')
f1_axes[1, 1] = dyn.pl.streamline_plot(adata, color='acceleration_umap', ax=f1_axes[1, 1], save_show_or_return='return')
plt.show()
_images/pancreatic_endocrinogenesis_16_0.png 
[14]:

# emulate ggplot2 plotting styple with black background
dyn.configuration.set_figure_params('dynamo', background='black')

[15]:

fig1, f1_axes = plt.subplots(ncols=2, nrows=2, constrained_layout=True, figsize=(12, 8))
f1_axes
f1_axes[0, 0] = dyn.pl.cell_wise_vectors(adata, color='umap_ddhodge_potential', pointsize=0.1, alpha = 0.7, ax=f1_axes[0, 0], quiver_length=6, quiver_size=6, save_show_or_return='return', background='black')
f1_axes[0, 1] = dyn.pl.grid_vectors(adata, color='speed_umap', ax=f1_axes[0, 1], quiver_length=12, quiver_size=12, save_show_or_return='return', background='black')
f1_axes[1, 0] = dyn.pl.streamline_plot(adata, color='divergence_pca', ax=f1_axes[1, 0], save_show_or_return='return', background='black')
f1_axes[1, 1] = dyn.pl.streamline_plot(adata, color='acceleration_umap', ax=f1_axes[1, 1], save_show_or_return='return', background='black')
plt.show()
_images/pancreatic_endocrinogenesis_18_0.png 
[16]:

progenitor = adata.obs_names[adata.obs.clusters.isin(['Ductal'])]
len(progenitor)

[16]:

916

[17]:

import numpy as np
dyn.pd.fate(adata, basis='umap', init_cells=np.random.choice(progenitor, 100), interpolation_num=100,  direction='forward',
   inverse_transform=False, average=False, cores=3)

[17]:

AnnData object with n_obs × n_vars = 3696 × 27998
    obs: 'clusters_coarse', 'clusters', 'S_score', 'G2M_score', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'Size_Factor', 'initial_cell_size', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'ntr', 'cell_cycle_phase', 'umap_ddhodge_div', 'umap_ddhodge_potential', 'curl_umap', 'divergence_umap', 'speed_umap', 'divergence_pca', 'acceleration_umap'
    var: 'highly_variable_genes', 'pass_basic_filter', 'log_m', 'score', 'log_cv', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics', 'use_for_transition'
    uns: 'clusters_coarse_colors', 'clusters_colors', 'day_colors', 'neighbors', 'pca', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'grid_velocity_umap', 'VecFld_umap', 'VecFld', 'grid_velocity_pca', 'VecFld_pca', 'fate_umap'
    obsm: 'X_pca', 'X_umap', 'X', 'cell_cycle_scores', 'velocity_umap', 'velocity_umap_SparseVFC', 'X_umap_SparseVFC', 'velocity_pca', 'velocity_pca_SparseVFC', 'X_pca_SparseVFC', 'acceleration_umap'
    layers: 'spliced', 'unspliced', 'X_spliced', 'X_unspliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'distances', 'connectivities', 'moments_con', 'pearson_transition_matrix', 'umap_ddhodge'

[18]:

%%capture
fig, ax = plt.subplots()
ax = dyn.pl.topography(adata, color='clusters', ax=ax, save_show_or_return='return')

[19]:

%%capture
instance = dyn.mv.StreamFuncAnim(adata=adata, ax=ax, color='clusters')

[20]:

import matplotlib
matplotlib.rcParams['animation.embed_limit'] = 2**128 # Ensure all frames will be embedded.

from matplotlib import animation
import numpy as np

anim = animation.FuncAnimation(instance.fig, instance.update, init_func=instance.init_background,
                                frames=np.arange(100), interval=100, blit=True)
from IPython.core.display import display, HTML
HTML(anim.to_jshtml()) # embedding to jupyter notebook.

[20]:

[21]:

%%capture
fig, ax = plt.subplots()
ax = dyn.pl.topography(adata, color='clusters', ax=ax, save_show_or_return='return')
dyn.mv.animate_fates(adata, color='clusters', basis='umap', n_steps=200, fig=fig, ax=ax,
                     save_show_or_return='save', logspace=True, max_time=None, save_kwargs={"filename": 'pancreas.gif'})