slingshot: Perform trajectory inference with Slingshot

In kstreet13/slingshot: Tools for ordering single-cell sequencing

Perform trajectory inference with Slingshot

Description

Perform trajectory inference with Slingshot

Perform trajectory inference by (1) identifying lineage structure with a cluster-based minimum spanning tree, and (2) constructing smooth representations of each lineage using simultaneous principal curves. This function wraps the getLineages and getCurves functions and is the primary function of the slingshot package.

Usage

slingshot(data, clusterLabels, ...)

## S4 method for signature 'matrix,character'
slingshot(
  data,
  clusterLabels,
  reducedDim = NULL,
  start.clus = NULL,
  end.clus = NULL,
  dist.method = "slingshot",
  use.median = FALSE,
  omega = FALSE,
  omega_scale = 1.5,
  times = NULL,
  shrink = TRUE,
  extend = "y",
  reweight = TRUE,
  reassign = TRUE,
  thresh = 0.001,
  maxit = 15,
  stretch = 2,
  approx_points = NULL,
  smoother = "smooth.spline",
  shrink.method = "cosine",
  allow.breaks = TRUE,
  ...
)

## S4 method for signature 'matrix,matrix'
slingshot(
  data,
  clusterLabels,
  reducedDim = NULL,
  start.clus = NULL,
  end.clus = NULL,
  dist.method = "slingshot",
  use.median = FALSE,
  omega = FALSE,
  omega_scale = 1.5,
  times = NULL,
  shrink = TRUE,
  extend = "y",
  reweight = TRUE,
  reassign = TRUE,
  thresh = 0.001,
  maxit = 15,
  stretch = 2,
  approx_points = NULL,
  smoother = "smooth.spline",
  shrink.method = "cosine",
  allow.breaks = TRUE,
  ...
)

## S4 method for signature 'SlingshotDataSet,ANY'
slingshot(data, clusterLabels, ...)

## S4 method for signature 'data.frame,ANY'
slingshot(data, clusterLabels, ...)

## S4 method for signature 'matrix,numeric'
slingshot(data, clusterLabels, ...)

## S4 method for signature 'matrix,factor'
slingshot(data, clusterLabels, ...)

## S4 method for signature 'matrix,ANY'
slingshot(data, clusterLabels, ...)

## S4 method for signature 'ClusterExperiment,ANY'
slingshot(
  data,
  clusterLabels,
  reducedDim = NULL,
  start.clus = NULL,
  end.clus = NULL,
  dist.method = "slingshot",
  use.median = FALSE,
  omega = FALSE,
  omega_scale = 1.5,
  times = NULL,
  shrink = TRUE,
  extend = "y",
  reweight = TRUE,
  reassign = TRUE,
  thresh = 0.001,
  maxit = 15,
  stretch = 2,
  approx_points = NULL,
  smoother = "smooth.spline",
  shrink.method = "cosine",
  allow.breaks = TRUE,
  ...
)

## S4 method for signature 'SingleCellExperiment,ANY'
slingshot(
  data,
  clusterLabels,
  reducedDim = NULL,
  start.clus = NULL,
  end.clus = NULL,
  dist.method = "slingshot",
  use.median = FALSE,
  omega = FALSE,
  omega_scale = 1.5,
  times = NULL,
  shrink = TRUE,
  extend = "y",
  reweight = TRUE,
  reassign = TRUE,
  thresh = 0.001,
  maxit = 15,
  stretch = 2,
  approx_points = NULL,
  smoother = "smooth.spline",
  shrink.method = "cosine",
  allow.breaks = TRUE,
  ...
)

Arguments

data	a data object containing the matrix of coordinates to be used for lineage inference. Supported types include matrix, SingleCellExperiment, SlingshotDataSet, and PseudotimeOrdering.
clusterLabels	each cell's cluster assignment. This can be a single vector of labels, or a #cells by #clusters matrix representing weighted cluster assignment. Either representation may optionally include a "-1" group meaning "unclustered."
...	Additional parameters to pass to scatter plot smoothing function, smoother.
reducedDim	(optional) the dimensionality reduction to be used. Can be a matrix or a character identifying which element of reducedDim(data) is to be used. If multiple dimensionality reductions are present and this argument is not provided, the first element will be used by default.
start.clus	(optional) character, indicates the starting cluster(s) from which lineages will be drawn.
end.clus	(optional) character, indicates which cluster(s) will be forced to be leaf nodes in the graph.
dist.method	(optional) character, specifies the method for calculating distances between clusters. Default is "slingshot", see createClusterMST for details.
use.median	logical, whether to use the median (instead of mean) when calculating cluster centroid coordinates.
omega	(optional) numeric, this granularity parameter determines the distance between every real cluster and the artificial cluster, .OMEGA. In practice, this makes omega the maximum allowable distance between two connected clusters. By default, omega = Inf. If omega = TRUE, the maximum edge length will be set to the median edge length of the unsupervised MST times a scaling factor (omega_scale, default = 3). This value is provided as a potentially useful rule of thumb for datasets with outlying clusters or multiple, distinct trajectories. See outgroup in createClusterMST.
omega_scale	(optional) numeric, scaling factor to use when omega = TRUE. The maximum edge length will be set to the median edge length of the unsupervised MST times omega_scale (default = 1.5). See outscale in createClusterMST.
times	numeric, vector of external times associated with either clusters or cells. See defineMSTPaths for details.
shrink	logical or numeric between 0 and 1, determines whether and how much to shrink branching lineages toward their average prior to the split (default = TRUE).
extend	character, how to handle root and leaf clusters of lineages when constructing the initial, piece-wise linear curve. Accepted values are 'y' (default), 'n', and 'pc1'. See 'Details' for more.
reweight	logical, whether to allow cells shared between lineages to be reweighted during curve fitting. If TRUE (default), cells shared between lineages will be iteratively reweighted based on the quantiles of their projection distances to each curve. See 'Details' for more.
reassign	logical, whether to reassign cells to lineages at each iteration. If TRUE (default), cells will be added to a lineage when their projection distance to the curve is less than the median distance for all cells currently assigned to the lineage. Additionally, shared cells will be removed from a lineage if their projection distance to the curve is above the 90th percentile and their weight along the curve is less than 0.1.
thresh	numeric, determines the convergence criterion. Percent change in the total distance from cells to their projections along curves must be less than thresh. Default is 0.001, similar to principal_curve.
maxit	numeric, maximum number of iterations (default = 15), see principal_curve.
stretch	numeric factor by which curves can be extrapolated beyond endpoints. Default is 2, see principal_curve.
approx_points	numeric, whether curves should be approximated by a fixed number of points. If FALSE (or 0), no approximation will be performed and curves will contain as many points as the input data. If numeric, curves will be approximated by this number of points (default = 150 or #cells, whichever is smaller). See 'Details' and principal_curve for more.
smoother	choice of scatter plot smoother. Same as principal_curve, but "lowess" option is replaced with "loess" for additional flexibility.
shrink.method	character denoting how to determine the appropriate amount of shrinkage for a branching lineage. Accepted values are the same as for kernel in density (default is "cosine"), as well as "tricube" and "density". See 'Details' for more.
allow.breaks	logical, determines whether curves that branch very close to the origin should be allowed to have different starting points.

Details

Given a reduced-dimensional data matrix n by p and a vector of cluster labels (or matrix of soft cluster assignments, potentially including a -1 label for "unclustered"), this function performs trajectory inference using a cluster-based minimum spanning tree on the clusters and simultaneous principal curves for smooth, branching paths.

The graph of this structure is learned by fitting a (possibly constrained) minimum-spanning tree on the clusters, plus the artificial cluster, .OMEGA, which is a fixed distance away from every real cluster. This effectively limits the maximum branch length in the MST to the chosen distance, meaning that the output may contain multiple trees.

Once the graph is known, lineages are identified in any tree with at least two clusters. For a given tree, if there is an annotated starting cluster, every possible path out of a starting cluster and ending in a leaf that isn't another starting cluster will be returned. If no starting cluster is annotated, one will be chosen by a heuristic method, but this is not recommended.

When there is only a single lineage, the curve-fitting algorithm is nearly identical to that of principal_curve. When there are multiple lineages and shrink > 0, an additional step is added to the iterative procedure, forcing curves to be similar in the neighborhood of shared points (ie., before they branch).

The approx_points argument, which sets the number of points to be used for each curve, can have a large effect on computation time. Due to this consideration, we set the default value to 150 whenever the input dataset contains more than that many cells. This setting should help with exploratory analysis while having little to no impact on the final curves. To disable this behavior and construct curves with the maximum number of points, set approx_points = FALSE.

The extend argument determines how to construct the piece-wise linear curve used to initiate the recursive algorithm. The initial curve is always based on the lines between cluster centers and if extend = 'n', this curve will terminate at the center of the endpoint clusters. Setting extend = 'y' will allow the first and last segments to extend beyond the cluster center to the orthogonal projection of the furthest point. Setting extend = 'pc1' is similar to 'y', but uses the first principal component of the cluster to determine the direction of the curve beyond the cluster center. These options typically have limited impact on the final curve, but can occasionally help with stability issues.

When shink = TRUE, we compute a percent shrinkage curve, w_l(t), for each lineage, a non-increasing function of pseudotime that determines how much that lineage should be shrunk toward a shared average curve. We set w_l(0) = 1 (complete shrinkage), so that the curves will always perfectly overlap the average curve at pseudotime 0. The weighting curve decreases from 1 to 0 over the non-outlying pseudotime values of shared cells (where outliers are defined by the 1.5*IQR rule). The exact shape of the curve in this region is controlled by shrink.method, and can follow the shape of any standard kernel function's cumulative density curve (or more precisely, survival curve, since we require a decreasing function). Different choices of shrink.method to have no discernable impact on the final curves, in most cases.

When reweight = TRUE, weights for shared cells are based on the quantiles of their projection distances onto each curve. The distances are ranked and converted into quantiles between 0 and 1, which are then transformed by 1 - q^2. Each cell's weight along a given lineage is the ratio of this value to the maximum value for this cell across all lineages.

Value

An object of class PseudotimeOrdering containing the pseudotime estimates and lineage assignment weights in the assays. The reducedDim and clusterLabels matrices will be stored in the cellData. Additionally, the metadata slot will contain an igraph object named mst, a list of parameter values named slingParams, a list of lineages (ordered sets of clusters) named lineages, and a list of principal_curve objects named curves.

References

Hastie, T., and Stuetzle, W. (1989). "Principal Curves." Journal of the American Statistical Association, 84:502-516.

Street, K., et al. (2018). "Slingshot: cell lineage and pseudotime inference for single-cell transcriptomics." BMC Genomics, 19:477.

Examples

data("slingshotExample")
rd <- slingshotExample$rd
cl <- slingshotExample$cl
pto <- slingshot(rd, cl, start.clus = '1')

# plotting
sds <- as.SlingshotDataSet(pto)
plot(rd, col = cl, asp = 1)
lines(sds, type = 'c', lwd = 3)

kstreet13/slingshot documentation built on April 6, 2023, 11:12 p.m.