Run CellRank analysis

CellRank is a toolkit for studying cellular dynamics using Markov state modeling.

Usage

RunCellRank(
  srt = NULL,
  assay_x = "RNA",
  layer_x = "counts",
  assay_y = c("spliced", "unspliced"),
  layer_y = "counts",
  adata = NULL,
  group.by = NULL,
  cores = 1,
  linear_reduction = NULL,
  nonlinear_reduction = NULL,
  basis = NULL,
  mode = "stochastic",
  fitting_by = "stochastic",
  magic_impute = FALSE,
  knn = 5,
  t = 2,
  min_shared_counts = 30,
  n_pcs = 30,
  n_neighbors = 30,
  stream_smooth = NULL,
  stream_density = 2,
  arrow_size = 5,
  arrow_length = 5,
  arrow_density = 0.5,
  calculate_velocity_genes = FALSE,
  denoise = FALSE,
  kinetics = FALSE,
  kernel_type = c("velocity", "pseudotime", "cytotrace"),
  time_key = "dpt_pseudotime",
  estimator_type = c("GPCCA", "CFLARE"),
  use_connectivity_kernel = TRUE,
  velocity_weight = 0.8,
  connectivity_weight = 0.2,
  softmax_scale = 4,
  n_macrostates = NULL,
  schur_method = c("krylov", "brandts"),
  n_cells_terminal = 10,
  show_plot = TRUE,
  save_plot = FALSE,
  plot_format = c("pdf", "png", "svg"),
  plot_dpi = 300,
  plot_prefix = "cellrank",
  legend.position = "on data",
  palette = "Chinese",
  palcolor = NULL,
  dirpath = "./cellrank",
  return_seurat = !is.null(srt),
  verbose = TRUE
)

Arguments

srt: A Seurat object. Default is NULL. If provided, adata will be ignored.
assay_x: Assay to convert as the main data matrix in the anndata object. Default is "RNA".
layer_x: Layer name for assay_x in the Seurat object. Default is "counts".
assay_y: Assays to convert as layers in the anndata object. Default is c("spliced", "unspliced").
layer_y: Layer names for the assay_y in the Seurat object. Default is "counts".
adata: An anndata object. Default is NULL.
group.by: Name of one or more meta.data columns to group (color) cells by.
cores: The number of cores to use for cellrank.
linear_reduction: The linear dimensionality reduction method to use. Options are "pca", "svd", "ica", "nmf", "mds", or "glmpca". Default is "pca".
nonlinear_reduction: The nonlinear dimensionality reduction method to use. Options are "umap", "umap-naive", "tsne", "dm", "phate", "pacmap", "trimap", "largevis", or "fr". Default is "umap".
basis: The basis to use for reduction, e.g., "UMAP".
mode: Velocity estimation models to use. Can be "deterministic", "stochastic", or "dynamical".
fitting_by: Method used to fit gene velocities for dynamical modeling. Default is "stochastic".
magic_impute: Flag indicating whether to perform magic imputation. Default is FALSE.
knn: The number of nearest neighbors for magic.MAGIC. Default is 5.
t: Power to which the diffusion operator is powered for magic.MAGIC. Default is 2.
min_shared_counts: Minimum number of counts (both unspliced and spliced) required for a gene. Default is 30.
n_pcs: Number of principal components to use for linear reduction. Default is 30.
n_neighbors: Number of neighbors to use for constructing the KNN graph. Default is 30.
stream_smooth: Multiplication factor for scale in Gaussian kernel around grid point.
stream_density: Controls the closeness of streamlines. When density = 2 (default), the domain is divided into a 60x60 grid, whereas density linearly scales this grid. Each cell in the grid can have, at most, one traversing streamline. Default is 2.
arrow_size: Size of arrows. Default is 5.
arrow_length: Length of arrows.
arrow_density: Amount of velocities to show.
calculate_velocity_genes: Boolean flag indicating whether to calculate velocity genes.
denoise: Boolean flag indicating whether to denoise.
kinetics: Boolean flag indicating whether to estimate RNA kinetics.
kernel_type: Type of kernel to use: "velocity" (default, requires spliced/unspliced), "pseudotime" (requires pre-computed pseudotime or auto-computes DPT), or "cytotrace" (auto-computes CytoTRACE score, suitable for RNA-only data).
time_key: Key in metadata for pseudotime. Used when kernel_type = "pseudotime". If the key doesn't exist, DPT pseudotime will be computed automatically. Default is "dpt_pseudotime".
estimator_type: Type of estimator to use: "GPCCA" (default) or "CFLARE". GPCCA provides coarse-grained analysis and Schur decomposition.
use_connectivity_kernel: Whether to combine the main kernel with ConnectivityKernel. Default is TRUE.
velocity_weight: Weight for the VelocityKernel when combining with ConnectivityKernel. Default is 0.8.
connectivity_weight: Weight for the ConnectivityKernel when combining with VelocityKernel. Default is 0.2. Weights are automatically normalized to sum to 1.0.
softmax_scale: Scaling parameter for softmax transformation of velocity kernel. Default is 4.
n_macrostates: Number of macrostates to compute. If NULL (default), automatically determined based on eigenvalue spectrum.
schur_method: Method for Schur decomposition: "krylov" or "brandts". Only used for GPCCA estimator.
n_cells_terminal: Minimum number of cells required for a state to be considered terminal. Default is 10.
show_plot: Whether to show the plot. Default is FALSE.
save_plot: Whether to save plots to files. Default is FALSE.
plot_format: Format for saved plots: "png" (default), "pdf", or "svg".
plot_dpi: Resolution (DPI) for saved plots. Default is 300.
plot_prefix: Prefix for saved plot filenames. Default is "cellrank".
legend.position: Position of legend in plots. Can be "on data", "right margin", "bottom right", etc. Default is "on data".
palette: Color palette name. Available palettes can be found in thisplot::show_palettes. Default is "Chinese".
palcolor: Custom colors used to create a color palette. Default is NULL.
dirpath: The directory to save the plots. Default is "./cellrank".
return_seurat: Whether to return a Seurat object instead of an anndata object. Default is TRUE.
verbose: Whether to print the message. Default is TRUE.

Value

Returns a Seurat object if return_seurat = TRUE or an anndata object with CellRank results stored in obsm, obs, and varm slots. The estimator and kernel objects are stored in srt@misc$cellrank.

Examples

if (FALSE) { # \dontrun{
data(pancreas_sub)
pancreas_sub <- standard_scop(pancreas_sub)
pancreas_sub <- RunCellRank(
  srt = pancreas_sub,
  group.by = "SubCellType"
)

CellDimPlot(
  pancreas_sub,
  group.by = "term_states_fwd",
  reduction = "umap",
  label = TRUE
)

FeatureDimPlot(
  pancreas_sub,
  features = "latent_time",
  reduction = "umap"
)

FeatureDimPlot(
  pancreas_sub,
  features = c("stochastic_confidence", "stochastic_length"),
  reduction = "umap"
)

CellDimPlot(
  pancreas_sub,
  group.by = "SubCellType",
  reduction = "UMAP",
  lineages = "cellrank_pseudotime",
  lineages_span = 0.1,
  lineages_trim = c(0.05, 0.95)
)

DynamicPlot(
  pancreas_sub,
  lineages = "cellrank_pseudotime",
  features = c("Arxes1", "Ncoa2"),
  group.by = "SubCellType"
)
} # }

Usage

Arguments

Value

See also

Examples