Package 'scAnnotatR'

caret_model

Description

Returns the caret model of the scAnnotatR object

Usage

caret_model(classifier)

Arguments

classifier

scAnnotatR object

Value

Classifier is the object returned by caret SVM learning process. More information about the caret package: https://topepo.github.io/caret/

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
caret_model(classifier_b)

---

cell_type

Description

Returns the cell type for the given classifier.

Usage

cell_type(classifier)

## S4 replacement method for signature 'scAnnotatR'
cell_type(classifier) <- value

Arguments

classifier	scAnnotatR object. The object is returned from the train_classifier function.
value	the new cell type

Value

cell type of object

scAnnotatR object with the new cell type

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
cell_type(classifier_b)

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
cell_type(classifier_b) <- "B cell"

---

Setter for cell_type. Change cell type of a classifier

Description

Setter for cell_type. Change cell type of a classifier

Usage

cell_type(classifier) <- value

Arguments

classifier	the classifier whose cell type will be changed
value	the new cell type

Value

the classifier with the new cell type

---

Internal functions of scAnnotatR package

Description

Check if a scAnnotatR object is valid

Train a classifier for a new cell type If cell type has a parent, only available for scAnnotatR object as parent cell classifying model.

Train a classifier for a new cell type If cell type has a parent, only available for scAnnotatR object as parent cell classifying model.

Train a classifier for a new cell type from expression matrix and tag If cell type has a parent, only available for scAnnotatR object as parent cell classifying model.

Preprocess Seurat object to produce expression matrix, tag, parent cell tag.

Preprocess Seurat object to produce expression matrix, tag, parent cell tag.

Testing process when test object is of type Seurat

Testing process when test object is of type SCE

Testing process from matrix and tag

This function ensures that parent classifiers are also selected.

Usage

checkObjectValidity(object)

checkCellTypeValidity(cell_type)

checkMarkerGenesValidity(marker_genes)

checkParentValidity(parent)

checkPThresValidity(p_thres)

checkCaretModelValidity(caret_model)

parent(classifier) <- value

## S4 replacement method for signature 'scAnnotatR'
parent(classifier) <- value

caret_model(classifier) <- value

## S4 replacement method for signature 'scAnnotatR'
caret_model(classifier) <- value

marker_genes(classifier) <- value

## S4 replacement method for signature 'scAnnotatR'
marker_genes(classifier) <- value

train_classifier_seurat(
  train_obj,
  cell_type,
  marker_genes,
  parent_cell = NA_character_,
  parent_classifier = NULL,
  path_to_models = "default",
  zscore = TRUE,
  seurat_tag_slot,
  seurat_parent_tag_slot = "predicted_cell_type",
  seurat_assay,
  seurat_slot,
  ambiguous_chars
)

train_classifier_sce(
  train_obj,
  cell_type,
  marker_genes,
  parent_cell = NA_character_,
  parent_classifier = NULL,
  path_to_models = "default",
  zscore = TRUE,
  sce_tag_slot,
  sce_parent_tag_slot = "predicted_cell_type",
  sce_assay,
  ambiguous_chars = NULL
)

train_classifier_from_mat(
  mat,
  tag,
  cell_type,
  marker_genes,
  parent_tag,
  parent_cell,
  parent_classifier,
  path_to_models,
  zscore,
  ambiguous_chars = NULL
)

preprocess_seurat_object(
  seurat_obj,
  seurat_assay,
  seurat_slot,
  seurat_tag_slot,
  seurat_parent_tag_slot
)

preprocess_sce_object(sce_obj, sce_assay, sce_tag_slot, sce_parent_tag_slot)

test_classifier_seurat(
  test_obj,
  classifier,
  target_cell_type = NULL,
  parent_classifier = NULL,
  path_to_models = "default",
  zscore = TRUE,
  seurat_tag_slot,
  seurat_parent_tag_slot = "predicted_cell_type",
  seurat_assay,
  seurat_slot,
  ambiguous_chars = NULL
)

test_classifier_sce(
  test_obj,
  classifier,
  target_cell_type = NULL,
  parent_classifier = NULL,
  path_to_models = "default",
  zscore = TRUE,
  sce_tag_slot,
  sce_parent_tag_slot = "predicted_cell_type",
  sce_assay,
  ambiguous_chars = NULL
)

test_classifier_from_mat(
  mat,
  tag,
  classifier,
  parent_tag,
  target_cell_type,
  parent_classifier,
  path_to_models,
  zscore,
  ambiguous_chars = NULL
)

classify_cells_seurat(
  classify_obj,
  classifiers = NULL,
  cell_types = "all",
  chunk_size = 5000,
  path_to_models = "default",
  ignore_ambiguous_result = FALSE,
  cluster_slot,
  seurat_assay,
  seurat_slot
)

classify_cells_sce(
  classify_obj,
  classifiers = NULL,
  cell_types = "all",
  chunk_size = 5000,
  path_to_models = "default",
  ignore_ambiguous_result = FALSE,
  sce_assay,
  cluster_slot = NULL
)

balance_dataset(mat, tag)

train_func(mat, tag)

transform_to_zscore(mat)

subset_models(model_list, model_names)

select_marker_genes(mat, marker_genes)

check_parent_child_coherence(
  mat,
  tag,
  pos_parent,
  parent_cell,
  cell_type,
  target_cell_type
)

filter_cells(mat, tag, ambiguous_chars = NULL)

construct_tag_vect(tag, cell_type)

process_parent_classifier(
  mat,
  parent_tag,
  parent_cell_type,
  parent_classifier,
  path_to_models,
  zscore
)

make_prediction(mat, classifier, pred_cells, ignore_ambiguous_result = TRUE)

simplify_prediction(meta.data, full_pred, classifiers)

verify_parent(mat, classifier, meta.data)

test_performance(mat, classifier, tag)

classify_clust(clusts, most_probable_cell_type)

download_data_file(verbose = FALSE)

Arguments

object	The request classifier to check.
cell_type	name of cell type
marker_genes	list of selected marker genes
parent	Classifier parent to check.
p_thres	Classifier probability threshold to check.
caret_model	Classifier to check.
classifier	classifier
value	the new classifier
train_obj	SCE object
parent_cell	name of parent cell type
parent_classifier	scAnnotatR object corresponding to classification model for the parent cell type
path_to_models	path to databases, or by default
zscore	boolean indicating the transformation of gene expression in object to zscore or not
seurat_tag_slot	string, name of annotation slot indicating cell tag/label in the testing object. Strings indicating cell types are expected in this slot. Expected values are string (A-Z, a-z, 0-9, no special character accepted) or binary/logical, 0/"no"/F/FALSE: not being new cell type, 1/"yes"/T/TRUE: being new cell type.
seurat_parent_tag_slot	string, name of tag slot in cell meta data indicating pre-assigned/predicted parent cell type. Default field is "predicted_cell_type". The slot must contain only string values.
seurat_assay	name of assay to use in Seurat object
seurat_slot	type of expression data to use in Seurat object. Some available types are: "counts", "data" and "scale.data".
ambiguous_chars	Vector of character (sequences) that if contained within a cell type mark this cell type as being ambiguous. If NULL default values are used. Charactes with a meaning in REGEX must be enclosed by []. F.e. "[+]". Default value is "/", ",", " -", " [+]", "[.]", " and ", " or ", "_or_", "-or-", "[(]" ,"[)]", "ambiguous"
sce_tag_slot	string, name of annotation slot indicating cell tag/label in the testing object. Strings indicating cell types are expected in this slot. Expected values are string (A-Z, a-z, 0-9, no special character accepted) or binary/logical, 0/"no"/F/FALSE: not being new cell type, 1/"yes"/T/TRUE: being new cell type.
sce_parent_tag_slot	string, name of tag slot in cell meta data indicating pre-assigned/predicted parent cell type. Default field is "predicted_cell_type". The slot must contain only string values.
sce_assay	name of assay to use in SCE object
mat	expression matrix
tag	tag of data
parent_tag	vector, named list indicating pre-assigned/predicted parent cell type
seurat_obj	Seurat object
sce_obj	Seurat object
test_obj	SCE object used for testing
target_cell_type	alternative cell types (in case of testing classifier)
classify_obj	the SCE object containing cells to be classified
classifiers	classifiers
cell_types	list of cell types containing models to be used for classification, only applicable if the models have been saved to package.
chunk_size	size of data chunks to be predicted separately. This option is recommended for large datasets to reduce running time. Default value at 5000, because smaller datasets can be predicted rapidly.
ignore_ambiguous_result	whether ignore ambigouous result
cluster_slot	name of slot in meta data containing cluster information, in case users want to have additional cluster-level prediction
model_list	A list of models
model_names	The names of the models to retain
pos_parent	a vector indicating parent classifier prediction
parent_cell_type	name of parent cell type
pred_cells	a whole prediction for all cells
meta.data	object meta data
full_pred	full prediction
clusts	cluster info
most_probable_cell_type	predicted cell type
verbose	logical indicating downloading the file or not

Value

TRUE if the classifier is valid or the reason why it is not

TRUE if the cell type is valid or the reason why it is not.

TRUE if the marker_genes is valid or the reason why it is not.

TRUE if the parent is valid or the reason why it is not.

TRUE if the p_thres is valid or the reason why it is not.

TRUE if the classifier is valid or the reason why it is not.

the classifier with the new parent.

scAnnotatR object with the new parent

the classifier with the new core caret model.

scAnnotatR object with the new trained classifier.

the classifier with the new marker genes

scAnnotatR object with the new marker genes.

scAnnotatR object

scAnnotatR object

caret trained model

a list containing: expression matrix of size n x m, n: genes, m: cells; a vector indicating cell type, and a vector containing parent cell type.

a list containing: expression matrix of size n x m, n: genes, m: cells; a vector indicating cell type, and a vector containing parent cell type.

result of testing process in form of a list, including predicted values, prediction accuracy at a probability threshold, and roc curve information.

result of testing process in form of a list, including predicted values, prediction accuracy at a probability threshold, and roc curve information.

model performance statistics

the input object with new slots in cells meta data New slots are: predicted_cell_type, most_probable_cell_type, slots in form of [cell_type]_p, [cell_type]_class, and clust_pred (if cluster_slot was provided).

the input object with new slots in cells meta data New slots are: predicted_cell_type, most_probable_cell_type, slots in form of [cell_type]_p, [cell_type]_class, and clust_pred (if cluster_slot was provided).

a list of balanced count matrix and corresponding tags of balanced count matrix

the classification model (caret object)

row wise center-scaled count matrix

The list containing the selected models

filtered matrix

list of adjusted tag

filtered matrix and corresponding tag

a binary vector for cell tag

list of cells which are positive to parent classifier

prediction

simplified prediction

applicable matrix

classifier performance

model list object

---

Classify cells from multiple models

Description

Classify cells from multiple models

Usage

classify_cells(
  classify_obj,
  assay,
  slot = NULL,
  classifiers = NULL,
  cell_types = "all",
  chunk_size = 5000,
  path_to_models = "default",
  ignore_ambiguous_result = FALSE,
  cluster_slot = "clusters"
)

Arguments

classify_obj	the object containing cells to be classified
assay	name of assay to use in classify_object
slot	type of expression data to use in classify_object. For Seurat object, some available types are: "counts", "data" and "scale.data".
classifiers	list of classification models. The model is obtained from train_classifier function or available in current working space. Users may test the model using test_classifier before using this function. If classifiers contain classifiers for sub cell types, classifiers for parent cell type must be indicated first in order to be applied before children classifiers. If classifiers is NULL, the method will use all classifiers in database.
cell_types	list of cell types containing models to be used for classification, only applicable if the models have been saved to package.
chunk_size	size of data chunks to be predicted separately. This option is recommended for large datasets to reduce running time. Default value at 5000, because smaller datasets can be predicted rapidly.
path_to_models	path to the folder containing the list of models. As default value, the pretrained models in the package will be used. If user has trained new models, indicate the folder containing the new_models.rda file.
ignore_ambiguous_result	return all ambiguous predictions (multiple cell types) to empty When this parameter turns to TRUE, most probably predicted cell types will be ignored.
cluster_slot	name of slot in meta data containing cluster information, in case users want to have additional cluster-level prediction

Value

the input object with new slots in cells meta data New slots are: predicted_cell_type, most_probable_cell_type, slots in form of [cell_type]_p, [cell_type]_class, and clust_pred (if cluster_slot was provided).

Examples

# load small example dataset
data("tirosh_mel80_example")

# train one classifier for one cell type, for ex, B cell
# define genes to use to classify this cell type
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")

# train the classifier
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B,
cell_type = "b cells", tag_slot = 'active.ident')

# do the same thing with other cell types, for example, T cells
selected_marker_genes_T = c("CD4", "CD8A", "CD8B")
set.seed(123)
classifier_t <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_T,
cell_type = "T cells", tag_slot = 'active.ident')

# create a list of classifiers
classifier_ls <- list(classifier_b, classifier_t)

# classify cells with list of classifiers
seurat.obj <- classify_cells(classify_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', classifiers = classifier_ls)

---

Delete model/branch from package

Description

Delete model/branch from package

Usage

delete_model(cell_type, path_to_models = tempdir())

Arguments

cell_type	string indicating the cell type of which the model will be removed from package Attention: deletion of a parent model will also delete all of child model.
path_to_models	path to the folder containing the list of models in which the to-be-deleted model is.

Value

no return value, but the model is deleted from database

Examples

# load small example dataset
data("tirosh_mel80_example")

# train a classifier
set.seed(123)
selected_marker_genes_T = c("CD4", "CD8A", "CD8B")
classifier_t <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_T, 
cell_type = "t cells", tag_slot = 'active.ident')

# save a classifier to system
save_new_model(new_model = classifier_t, path_to_models = tempdir(), 
               include.default = FALSE)

# delete classifier from system
delete_model("t cells", path_to_models = tempdir())

---

Load classifiers from databases

Description

Load classifiers from databases

Usage

load_models(path_to_models)

Arguments

path_to_models

path to databases, or by default

Value

list of classifiers

---

marker_genes

Description

Returns the set of marker genes for the given classifier.

Usage

marker_genes(classifier)

Arguments

classifier

scAnnotatR object

Value

Applied marker genes of object

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
marker_genes(classifier_b)

---

p_thres

Description

Returns the probability threshold for the given classifier.

Usage

p_thres(classifier)

## S4 replacement method for signature 'scAnnotatR'
p_thres(classifier) <- value

Arguments

classifier	scAnnotatR object. The object is returned from the train_classifier function.
value	the new threshold

Value

Predicting probability threshold of object

scAnnotatR object with the new threshold.

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
p_thres(classifier_b)

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
classifier_b_test <- test_classifier(classifier = classifier_b, 
test_obj = tirosh_mel80_example, assay = 'RNA', slot = 'counts', 
tag_slot = 'active.ident')
# assign a new threhold probability for prediction
p_thres(classifier_b) <- 0.4

---

Setter for predicting probability threshold

Description

Setter for predicting probability threshold

Usage

p_thres(classifier) <- value

Arguments

classifier	the classifier whose predicting probability threshold will be changed
value	the new threshold

Value

classifier with the new threshold.

---

parent

Description

Returns the parent of the cell type corresponding to the given classifier.

Usage

parent(classifier)

Arguments

classifier

scAnnotatR object

Value

Parent model of object

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
parent(classifier_b)

---

Plant tree from list of models

Description

Plant tree from list of models

Usage

plant_tree(path_to_models = "default")

Arguments

path_to_models

list of models. If not provided, list of default pretrained models in the package will be used.

Value

tree structure and plot of tree

Examples

# to create the tree of classifiers 
# (in this example, based on default classifiers)
plant_tree()

---

Plot roc curve

Description

Plot roc curve

Usage

plot_roc_curve(test_result)

Arguments

test_result

result of test_classifier function

Value

ggplot2 roc curve

Examples

# load small example dataset
data("tirosh_mel80_example")

# train a classifier, for ex: B cell
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B,
cell_type = "b cells", tag_slot = 'active.ident')

classifier_b_test <- test_classifier(classifier = classifier_b,
test_obj = tirosh_mel80_example, assay = 'RNA', slot = 'counts',
tag_slot = 'active.ident', target_cell_type = c("B cell"))

# run plot curve on the test result
roc_curve <- plot_roc_curve(test_result = classifier_b_test)

---

Save a model to the package

Description

Save a model to the package

Usage

save_new_model(new_model, include.default = TRUE, path_to_models = tempdir())

Arguments

new_model	new model to be added into the classification tree
include.default	whether include the default models of the package in the list of new trained models or not. If users further want to classify cells, they can only use default pretrained model list or their new model list. Including default models in new trained models helps users using both of them once. In addition, default pretrained models of the package cannot be changed or removed. This can be done with the new trained model list.
path_to_models	path to the folder containing the list of new models.

Value

no return value, but the model is now saved to database

Examples

# load small example dataset
data("tirosh_mel80_example")

# train classifier
selected_marker_genes_T = c("CD4", "CD8A", "CD8B")
set.seed(123)
classifier_t <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_T, 
cell_type = "t cells", tag_slot = 'active.ident')

# save the trained classifier to system 
# test classifier can be used before this step
# Note: We do not include the default models here to runtime of the example
save_new_model(new_model = classifier_t, path_to_models = tempdir(), 
               include.default = FALSE)

# verify if new model has been saved
print(names(load(file.path(tempdir(), "new_models.rda"))))
delete_model("t cells")

---

scAnnotatR class.

Description

This class is returned by the train_classifier function. Generally, scAnnotatR objects are never created directly.

Usage

scAnnotatR(cell_type, caret_model, marker_genes, p_thres, parent)

scAnnotatR(cell_type, caret_model, marker_genes, p_thres, parent)

Arguments

cell_type	character. Name of the cell type.
caret_model	list. Trained model returned by caret train function.
marker_genes	vector/character containing marker genes used for the training.
p_thres	numeric. Probability threshold for the cell type to be assigned for a cell.
parent	character. Parent cell type.

Value

A scAnnotatR object.

Slots

cell_type

character. Name of the cell type.

caret_model

list. Trained model returned by caret train function.

marker_genes

vector/character containing marker genes used for the training.

p_thres

numeric. Probability threshold for the cell type to be assigned for a cell.

parent

character. Parent cell type.

Examples

# load small example dataset
data("tirosh_mel80_example")

# train a classifier, for ex: B cell
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')

classifier_b

---

Show object

Description

Show object

Usage

## S4 method for signature 'scAnnotatR'
show(object)

Arguments

object

scAnnotatR object

Value

print to console information about the object

Examples

data("tirosh_mel80_example")
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B, 
cell_type = "B cells", tag_slot = 'active.ident')
classifier_b

---

Testing process.

Description

Testing process.

Usage

test_classifier(
  classifier,
  test_obj,
  assay,
  slot = NULL,
  tag_slot,
  target_cell_type = NULL,
  parent_classifier = NULL,
  parent_tag_slot = "predicted_cell_type",
  path_to_models = "default",
  zscore = TRUE,
  ambiguous_chars = NULL
)

## S4 method for signature 'scAnnotatR'
test_classifier(
  classifier,
  test_obj,
  assay,
  slot = NULL,
  tag_slot,
  target_cell_type = NULL,
  parent_classifier = NULL,
  parent_tag_slot = "predicted_cell_type",
  path_to_models = "default",
  zscore = TRUE,
  ambiguous_chars = NULL
)

Arguments

classifier	scAnnotatR classification model
test_obj	object that can be used for testing
assay	name of assay to use in test_object
slot	type of expression data to use in test_object. For Seurat object, some available types are: "counts", "data" and "scale.data". Ignore this if test_obj is SingleCellExperiment object.
tag_slot	string, name of annotation slot indicating cell tag/label in the testing object. Strings indicating cell types are expected in this slot. Expected values are string (A-Z, a-z, 0-9, no special character accepted) or binary/logical, 0/"no"/F/FALSE: not being new cell type, 1/"yes"/T/TRUE: being new cell type.
target_cell_type	vector indicating other cell types than cell labels that can be considered as the main cell type in classifier, for example, c("plasma cell", "b cell", "b cells", "activating b cell"). Default as NULL.
parent_classifier	scAnnotatR object corresponding to classification model for the parent cell type
parent_tag_slot	string, name of tag slot in cell meta data indicating pre-assigned/predicted parent cell type. Default field is "predicted_cell_type". The slot must contain only string values.
path_to_models	path to the folder containing the list of models. As default, the pretrained models in the package will be used. If user has trained new models, indicate the folder containing the new_models.rda file.
zscore	boolean, whether gene expression is transformed to zscore
ambiguous_chars	List of characters to indicate ambiguous cells. For more details see filter_cells.

Value

result of testing process in form of a list, including predicted values, prediction accuracy at a probability threshold, and roc curve information.

Note

Only one cell type is expected for each cell. Ambiguous cell type, such as: "T cells/NK cells/ILC", will be ignored. Subtypes used in testing model for parent cell types can be indicated as parent cell type, or can be indicated in target_cell_type. For example, when testing for B cells, plasma cells can be annotated as B cells, or target_cell_type is set c("plasma cells").

Examples

# load small example dataset
data("tirosh_mel80_example")

# train the classifier
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B,
cell_type = "b cells", tag_slot = 'active.ident')

# test the classifier, target cell type can be in other formats or
# alternative cell type that can be considered as the classified cell type
classifier_b_test <- test_classifier(classifier = classifier_b,
test_obj = tirosh_mel80_example, assay = 'RNA', slot = 'counts',
tag_slot = 'active.ident', target_cell_type = c("B cell"))
classifier_b_test

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A Seurat Object Sample

Description

An example Seurat object shipped with the package as an example data. The expression data was originally from the dataset GSE72056, with samples corresponding to patient CY80. The Seurat object was then adapted to be used in scAnnotatR.

Usage

tirosh_mel80_example

Format

a Seurat object

Author(s)

Itay Tirosh, 2016-04-05

Source

WEIZMANN INSTITUTE OF SCIENCE

---

Train cell type classifier

Description

Train a classifier for a new cell type. If cell type has a parent, only available for scAnnotatR object as parent cell classifying model.

Usage

train_classifier(
  train_obj,
  assay,
  slot = NULL,
  cell_type,
  marker_genes,
  tag_slot,
  parent_cell = NA_character_,
  parent_tag_slot = "predicted_cell_type",
  parent_classifier = NULL,
  path_to_models = "default",
  zscore = TRUE,
  ambiguous_chars = NULL
)

Arguments

train_obj	object that can be used for training the new model. Seurat object or SingleCellExperiment object is supported. If the training model has parent, parent_tag_slot may have been indicated. This field would have been filled out automatically if user precedently run classify_cells function. If no (predicted) cell type annotation provided, the function can be run if 1- parent_cell or 2- parent_classifier is provided.
assay	name of assay to use in training object.
slot	type of expression data to use in training object, omitted if train_obj is SingleCellExperiment object.
cell_type	string indicating the name of the subtype This must exactly match cell tag/label if cell tag/label is a string.
marker_genes	list of marker genes used for the new training model
tag_slot	string, name of slot in cell meta data indicating cell tag/label in the training object. Strings indicating cell types are expected in this slot. For Seurat object, default value is "active.ident". Expected values are string (A-Z, a-z, 0-9, no special character accepted) or binary/logical, 0/"no"/F/FALSE: not being new cell type, 1/"yes"/T/TRUE: being new cell type.
parent_cell	string indicated the name of the parent cell type, if parent cell type classifier has already been saved in model database. Adjust path_to_models for exact database.
parent_tag_slot	string, name of a slot in cell meta data indicating assigned/predicted cell type. Default is "predicted_cell_type". This slot would have been filled automatically if user have called classify_cells function. The slot must contain only string values.
parent_classifier	classification model for the parent cell type
path_to_models	path to the folder containing the model database. As default, the pretrained models in the package will be used. If user has trained new models, indicate the folder containing the new_models.rda file.
zscore	whether gene expression in train_obj is transformed to zscore
ambiguous_chars	List of characters to indicate ambiguous cells. For more details see filter_cells.

Value

scAnnotatR object

Note

Only one cell type is expected for each cell in object. Ambiguous cell type, such as: "T cells/NK cells/ILC", will be ignored from training. Subtypes used in training model for parent cell types must be indicated as parent cell type. For example, when training for B cells, plasma cells must be annotated as B cells in order to be used.

Examples

# load small example dataset
data("tirosh_mel80_example")

# this dataset already contains pre-defined cell labels
table(Seurat::Idents(tirosh_mel80_example))

# define genes to use to classify this cell type (B cells in this example)
selected_marker_genes_B = c("CD19", "MS4A1", "CD79A")

# train the classifier, the "cell_type" argument must match
# the cell labels in the data, except upper/lower case
set.seed(123)
classifier_b <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', marker_genes = selected_marker_genes_B,
cell_type = "b cells", tag_slot = 'active.ident')

# classify cell types using B cell classifier,
# a test classifier process may be used before applying the classifier
tirosh_mel80_example <- classify_cells(classify_obj = tirosh_mel80_example,
classifiers = c(classifier_b), assay = 'RNA', slot = 'counts')

# tag all cells that are plasma cells (random example here)
tirosh_mel80_example[['plasma_cell_tag']] <- c(rep(1, 80), rep(0, 400))

# set new marker genes for the subtype
p_marker_genes = c("SDC1", "CD19", "CD79A")

# train the classifier, the "B cell" classifier is used as parent.
# This means, only cells already classified as "B cells" will be evaluated.
# the "tag_slot" parameter tells the classifier to use this cell meta data
# for the training process.
set.seed(123)
plasma_classifier <- train_classifier(train_obj = tirosh_mel80_example,
assay = 'RNA', slot = 'counts', cell_type = 'Plasma cell',
marker_genes = p_marker_genes, tag_slot = 'plasma_cell_tag',
parent_classifier = classifier_b)

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