run.CSIDE.regions: Runs CSIDE on a 'RCTD' object for DE across multiple discrete...

View source: R/CSIDE.R

run.CSIDE.regionsR Documentation

Runs CSIDE on a RCTD object for DE across multiple discrete regions

Description

Identifies cell type specific differential expression (DE) across multiple discrete regions The design matrix contains for each region a column of 0s and 1s representing membership in that region. Uses maximum likelihood estimation to estimate DE and standard errors for each gene and each cell type. Selects genes with significant nonzero DE. Tests for differences in gene expression across regions.

Usage

run.CSIDE.regions(
  myRCTD,
  region_list,
  cell_types = NULL,
  cell_type_threshold = 125,
  gene_threshold = 5e-05,
  doublet_mode = T,
  weight_threshold = NULL,
  sigma_gene = T,
  PRECISION.THRESHOLD = 0.05,
  cell_types_present = NULL,
  fdr = 0.01,
  test_genes_sig = T,
  logs = F,
  log_fc_thresh = 0.4,
  test_error = F
)

Arguments

myRCTD

an RCTD object with annotated cell types e.g. from the run.RCTD function.

region_list

a list of character vectors, where each vector contains pixel names, or barcodes, for a single region. These pixel names should be a subset of the pixels in the SpatialRNA object

cell_types

the cell types used for CSIDE. If null, cell types will be chosen with aggregate occurences of at least 'cell_type_threshold', as aggregated by aggregate_cell_types

cell_type_threshold

(default 125) min occurence of number of cells for each cell type to be used, as aggregated by aggregate_cell_types

gene_threshold

(default 5e-5) minimum average normalized expression required for selecting genes

doublet_mode

(default TRUE) if TRUE, uses RCTD doublet mode weights. Otherwise, uses RCTD full mode weights

weight_threshold

(default NULL) the threshold of total normalized weights across all cell types in cell_types per pixel to be included in the model. Default 0.99 for doublet_mode or 0.8 for full_mode.

sigma_gene

(default TRUE) if TRUE, fits gene specific overdispersion parameter. If FALSE, overdispersion parameter is same across all genes.

PRECISION.THRESHOLD

(default 0.05) for checking for convergence, the maximum parameter change per algorithm step

cell_types_present

cell types (a superset of 'cell_types') to be considered as occuring often enough to consider for gene expression contamination during the step filtering out marker genes of other cell types.

fdr

(default 0.01) false discovery rate for hypothesis testing

test_genes_sig

(default TRUE) logical controlling whether genes will be tested for significance

logs

(default FALSE) if TRUE, writes progress to logs/de_logs.txt

log_fc_thresh

(default 0.4) the natural log fold change cutoff for differential expression

test_error

(default FALSE) if TRUE, exits after testing for error messages without running CSIDE. If set to TRUE, this can be used to quickly evaluate if CSIDE will run without error.

Value

an RCTD object containing the results of the CSIDE algorithm. Contains objects de_results, which contain the results of the CSIDE algorithm including 'gene_fits', which contains the results of fits on individual genes, in addition 'sig_gene_list', a list, for each cell type, of significant genes detected by CSIDE. Additionally, the object contains 'internal_vars_de' a list of variables that are used internally by CSIDE


dmcable/RCTD documentation built on Feb. 24, 2024, 11:03 p.m.