smooth_gene

Returns the smoothed expression of the given gene, based on a GAM fit to the normalised expression.

Usage

smooth_gene(sce, gene, pseudotime_slot = "slingPseudotime_1", knots = 10)

Arguments

sce

SCE to do the calculations on.

gene

The name of the gene to smooth

pseudotime_slot

The slot in the SCE object containing pseudotime

knots

The number of knots to use when fitting the GAM

Value

Smoothed Gene Expression over pseudotime

Examples

ncells <- 70
ngenes <- 100
# Each gene should have mean around its gene number
counts <- c()
for (i in seq_len(ngenes)) {
    counts <- c(counts, dnorm(seq_len(ncells), mean = (ncells / i), sd = 1))
}

counts_matrix <- matrix(
    counts,
    ncol = ncells,
    nrow = ngenes
)
sce <- SingleCellExperiment::SingleCellExperiment(assays = list(
    counts = counts_matrix * 3,
    normcounts = counts_matrix,
    logcounts = log(counts_matrix)
))
colnames(sce) <- paste0("cell", seq_len(ncells))
rownames(sce) <- paste0("gene", seq_len(ngenes))
sce$cell_type <- c(
    rep("celltype_1", ncells / 2),
    rep("celltype_2", ncells / 2)
)

sce$pseudotime <- seq_len(ncells) - 1
genelist <- rownames(sce)

# calculate_gene_peakedness
gene_peakedness <- calculate_gene_peakedness(
    sce,
    pseudotime_slot = "pseudotime"
)
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Fitting terminated with step failure - check results carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Fitting terminated with step failure - check results carefully
#> Warning: Fitting terminated with step failure - check results carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully
#> Warning: Iteration limit reached without full convergence - check carefully

head(gene_peakedness)
#>      gene peak_pseudotime mean_in_window mean_out_window        ratio
#> 100 gene1           69.00   1.060616e-24     0.037863935 2.801125e-23
#> 51  gene2           35.19   4.680824e-02     0.029379725 1.593216e+00
#> 27  gene3           18.63   4.327921e-02     0.014445463 2.996042e+00
#> 20  gene4           13.80   4.563543e-02     0.006531685 6.986777e+00
#> 1   gene5            0.69   7.978846e-02     0.004138929 1.927756e+01
#> 5   gene6            3.45   3.456725e-02     0.006114805 5.653041e+00
#>     window_start window_end deviance_explained
#> 100        65.55      72.45         0.02680534
#> 51         31.74      38.64         0.01912609
#> 27         15.18      22.08         0.04505152
#> 20         10.35      17.25         0.58156238
#> 1          -2.76       4.14         0.21032129
#> 5           0.00       6.90         0.14902599

# plot_gene_peakedness
plot_gene_peakedness(sce, gene_peakedness, "gene20",
    pseudotime_slot = "pseudotime"
)
#> Warning: Iteration limit reached without full convergence - check carefully


# smooth_gene
smoothed_gene20 <- smooth_gene(
    sce, "gene20",
    pseudotime_slot = "pseudotime"
)
#> Warning: Iteration limit reached without full convergence - check carefully
head(smoothed_gene20)
#>            1            2            3            4            5            6 
#> 2.220446e-16 2.220446e-16 2.220446e-16 2.220446e-16 2.220446e-16 2.220446e-16 

# Select best spread of genes
genes_to_use <- gene_peakedness_spread_selection(sce, gene_peakedness,
    genes_per_bin = 2, n_gene_bins = 1, pseudotime_slot = "pseudotime"
)

print(genes_to_use)
#> [1] "gene30" "gene40"
plot(
    x = gene_peakedness[
        gene_peakedness$gene %in% genes_to_use, "peak_pseudotime"
    ],
    y = gene_peakedness[gene_peakedness$gene %in% genes_to_use, "ratio"]
)