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ontoProc: Ontology interfaces for Bioconductor, with focus on cell type identification
In ontoProc: processing of ontologies of anatomy, cell lines, and so on
Introduction
The ambitions of collaborative single cell biology
will only be achieved through the coordinated efforts
of many groups, to help clarify cell types
and dynamics in an array of functional and
environmental contexts. The use of formal ontology in
this pursuit is well-motivated and research progress has
already been substantial.
@Bakken2017 discuss "strategies for standardized cell type
representations based on the data outputs from [high-content
flow cytometry and single cell RNA sequencing], including
'context annotations' in the form of standardized experiment
metadata about the specimen source analyzed and marker
genes that serve as the most useful features
in machine learning-based cell type classification models."
@Aevermann2018 describe how the FAIR principles
can be implemented using statistical identification of necessary
and sufficient conditions for determining cell class membership.
They propose that Cell Ontology can be transformed
to a broadly usable knowledgebase through the incorporation
of accurate marker gene signatures for cell classes.
In this vignette, we review key concepts and tasks required to
make progress in the adoption and application of ontological discipline
in Bioconductor-oriented data analysis.
We'll start by setting up some package attachments and
ontology objects.
suppressPackageStartupMessages({
library(knitr)
library(ontoProc)
go = getGeneOnto()
cl = getCellOnto()
pr = getPROnto()
library(ontologyPlot)
library(BiocStyle)
})
Scope of package
The following table describes the resources available with get*
commands defined in ontoProc.
kable(packDesc2019)
Methods
Conceptual overview of ontology with cell types
Definitions, semantics. For concreteness, we provide some definitions and examples.
We use ontology to denote the systematic organization
of terminology used in a conceptual domain. The
Cell Ontology is a graphical data structure with
carefully annotated terms as nodes and conventionally defined
semantic relationships among terms serving as edges. As
an example, lung ciliated cell has URI \url{http://purl.obolibrary.org/obo/CL_1000271}.
This URI includes a fixed-length identifier CL_1000271 with
unambiguous interpretation wherever it is encountered. There is
a chain of relationships from lung ciliated cell
up through ciliated cell, then native cell, then
cell, each possessing its own URI and related
interpretive metadata. The relationship connecting the more precise
to the less precise term in this chain
is denoted SubclassOf. Ciliated cell is equivalent to
a native cell that has plasma membrane part
cilium. Semantic characteristics of terms and relationships are
used to infer relationships among terms that may
not have relations directly specified in available ontologies.
Barriers to broad adoption. Given the wealth of material available in biological
ontologies, it is somewhat surprising that formal annotation
is so seldom used in practice. Barriers to
more common use of ontology in data annotation
include: (i) Non-existence of exact matching between intended
term and terms available in ontologies of interest.
(ii) The practical problem of decoding ontology identifiers.
A GO tag or CL tag is excellent
for programming, but it is clumsy to co-locate
with the tag the associated natural language term
or phrase. (iii) Likelihood of disagreement of suitability
of terms for conditions observed at the boundaries
of knowledge. To help cope with the first
of these problems, Bioconductor's ontologyProc package
includes a function liberalMap
which will search an ontology for terms lexically
close to some target term or phrase. The
second problem can be addressed with more elaborate
data structures for variable annotation and programming in
R, and the third problem will diminish in
importance as the value of ontology adoption becomes
manifest in more applications.
Class vs. instance. It is important to distinguish the practice of
designing and maintaining ontologies from the use of
ontological class terms to annotate instances of the
concepts. The combination of an ontology and a
set of annotated instances is called a knowledge
base. To illustrate some of the salient distinctions
here, consider the cell line called A549, which
is established from a human lung adenocarcinoma sample.
There is no mention of A549 in the
Cell Ontology. However, A549 is present in the
EBI Experimental Factor Ontology as a subclass of
the "Homo sapiens cell line" class. Presumably this
is because A549 is a class of cells
that are widely used experimentally, and this cell
line constitutes a concept deserving of mapping in
the universe of experimental factors. In the universe
of concepts related to cell structure and function
per se, A549 is an individual that can
be characterized through possession of or lack of
properties enumerated in Cell Ontology, but it is
not deserving of inclusion in that ontology.
Illustration in a single-cell RNA-seq dataset
The 10X Genomics corporation has distributed
a dataset on results of sequencing 10000 PBMC from a healthy donor
\url{https://support.10xgenomics.com/single-cell-gene-expression/datasets}.
Subsets of the data are used in tutorials for the Seurat analytical
suite (@butler).
Labeling PBMC in the Seurat tutorial
One result of the tutorial analysis of the 3000 cell subset is a table
of cell types and expression-based markers of cell identity. The
first three columns of the table below are from
concluding material in the Seurat tutorial;
the remaining columns are created by "manual" matching
between the Seurat terms and terms found in Cell Ontology.
kable(stab <- seur3kTab())
Relationships asserted in the Cell Ontology
Given the informally selected tags in the table above, we can
sketch the Cell Ontology graph connecting the associated
cell types. The ontoProc package adds functionality to
ontologyPlot with make_graphNEL_from_ontology_plot. This
allows use of all Rgraphviz and igraph visualization facilities
for graphs derived from ontology structures.
Here we display the PBMC cell sets reported in the Seurat tutorial.
library(ontoProc)
cl = getCellOnto()
onto_plot2(cl, stab$tag)
Molecular features asserted in the Cell Ontology
The CLfeats function traces relationships and
properties from a given Cell Ontology class.
Briefly, each class can assert that it is the
intersection_of other classes, and
has_part, lacks_part, has_plasma_membrane_part,
lacks_plasma_membrane_part can be asserted as
relationships holding between cell type instances
and cell components. The components are often cross-referenced
to Protein Ontology or Gene Ontology. When the Protein Ontology
component has a synonym for which an HGNC symbol is provided, that
symbol is retrieved by CLfeats. Here we obtain the listing
for a mature CD1a-positive dermal dendritic cell.
kable(CLfeats(cl, "CL:0002531"))
The ctmarks function starts a shiny app that generates
tables of this sort for selected cell types.
Mapping from gene 'presence/role' to cell type
The sym2CellOnto function helps find mention of
given gene symbols in properties or parts of cell types.
kable(sdf <- as.data.frame(sym2CellOnto("ITGAM", cl, pr)))
table(sdf$cond)
kable(as.data.frame(sym2CellOnto("FOXP3", cl, pr)))
Adding terms to ontology_index structures to 'extend' Cell Ontology
The task of extending an ontology is partly bureaucratic in
nature and depends on a collection of endorsements and updates
to centralized information structures. In order to permit
experimentation with interfaces and new content that may
be quite speculative, we include an approach to combining new
ontology 'terms' of structure similar to those endorsed in
Cell Ontology, to ontologyIndex-based ontology_index
instances.
Use case: a set of cell types defined by "diagonal expression"
For a demonstration, we consider the discussion in
@Bakken2017, of a 'diagonal' expression pattern
defining a group of novel cell types. A set of genes
is identified and cells are distinguised by expressing
exactly one gene from the set.
The necessary information is collected in a vector.
The vector is the set of genes, the name of element i
is the tag to be associated with the type of cell that expresses gene i
and does not express any other gene in the set.
sigels = c("CL:X01"="GRIK3", "CL:X02"="NTNG1", "CL:X03"="BAGE2",
"CL:X04"="MC4R", "CL:X05"="PAX6", "CL:X06"="TSPAN12",
"CL:X07"="hSHISA8", "CL:X08"="SNCG", "CL:X09"="ARHGEF28",
"CL:X10"="EGF")
A data.frame defining the cell types and their properties
The cyclicSigset function produces a data.frame instance
connecting cell types with the genes expressed or unexpressed.
cs = cyclicSigset(sigels)
dim(cs)
cs[c(1:5,9:13),]
table(cs$cond)
It is expected that a tabular layout like this will suffice to
handle general situations of cell type definition.
Translating the data.frame elements to OBO Term instances
The most complicated aspect of novel OBO term construction is the
proper specifications of relationships with existing ontology components.
A prolog that is mostly shared by all terms is generated programmatically
for the diagonal pattern task.
makeIntnProlog = function(id, ...) {
# make type-specific prologs as key-value pairs
c(
sprintf("id: %s", id),
sprintf("name: %s-expressing cortical layer 1 interneuron, human", ...),
sprintf("def: '%s-expressing cortical layer 1 interneuron, human described via RNA-seq observations' [PMID 29322913]", ...),
"is_a: CL:0000099 ! interneuron",
"intersection_of: CL:0000099 ! interneuron")
}
The ldfToTerms API uses this to create a set of strings that can be parsed
as a term.
pmap = c("hasExp"="has_expression_of", lacksExp="lacks_expression_of")
head(unlist(tms <- ldfToTerms(cs, pmap, sigels, makeIntnProlog)), 20)
The content in tms can then be appended to the content of the Cell Ontology cl.obo as
text for import with ontologyIndex::get_OBO.
Subsetting SingleR resources using ontological mapping
A data.frame mapping from informal to formal terms
Aaron Lun has produced a mapping from informal terms used in the
Human Primary Cell Atlas to Cell Ontology tags. We provisionally
include a copy of this mapping in ontoProc:
hpca_map = read.csv(system.file("extdata/hpca.csv", package="ontoProc"), strings=FALSE)
head(hpca_map)
We will rename columns of this map for convenience of our bind_formal_tags method.
names(hpca_map) = c("informal", "formal") # obligatory for now
Binding formal tags to the HPCA data
I am turning this code off for now because there is no
standard approach to getting the mapping from the SummarizedExperment
yet. When SingleR merges the 'standardized' branch, this will come back.
Let's retrieve the HPCA data from SingleR:
library(SingleCellExperiment)
library(celldex)
hpca_sce = HumanPrimaryCellAtlasData()
Now bind the formal tags:
hpca_sce = bind_formal_tags(hpca_sce, "label.fine", hpca_map)
length(unique(hpca_sce$label.ont))
We don't check for failed mappings:
length(xx <- which(is.na(hpca_sce$label.ont)))
if (length(xx)>0) print(colData(hpca_sce)[xx,])
sum(hpca_sce$label.ont == "", na.rm=TRUE) # iPS and BM
Subsetting using the class hierarchy of Cell Ontology
cell_onto = ontoProc::getCellOnto()
hpca_mono = subset_descendants( hpca_sce, cell_onto, "^monocyte$" )
table(hpca_mono$label.fine)
table(hpca_mono$label.ont) # not much diversity
hpca_tcell = subset_descendants( hpca_sce, cell_onto, "^T cell$" )
table(hpca_tcell$label.fine)
table(hpca_tcell$label.ont) #
uu = unique(hpca_tcell$label.ont)
onto_plot2(cell_onto, uu)
Related tools
Inference on the identities of cells assayed in
a single cell transcriptomics experiment can be performed
using the Bioconductor celaref package. This package includes
a number of reference data resources providing whole-transcriptome
profiles of cells of known and unknown type.
An approach to systematically structuring data on cell-type
signatures, and conducting inference on cell types in
new experiments, is provided in the Hancock package,
under development.
A CRAN package that is very useful for
R programming with ontologies is ontologyIndex @Westbury2015. This
provides easily used functions for parsing ontologies in
the OBO format and for performing basic queries
on text fields and list structures.
References
Try the ontoProc package in your browser
Any scripts or data that you put into this service are public.
ontoProc documentation built on Nov. 8, 2020, 4:49 p.m.
R Package Documentation
Browse R Packages
We want your feedback!
Note that we can't provide technical support on individual packages. You should contact the package authors for that.
Introduction
The ambitions of collaborative single cell biology will only be achieved through the coordinated efforts of many groups, to help clarify cell types and dynamics in an array of functional and environmental contexts. The use of formal ontology in this pursuit is well-motivated and research progress has already been substantial.
@Bakken2017 discuss "strategies for standardized cell type representations based on the data outputs from [high-content flow cytometry and single cell RNA sequencing], including 'context annotations' in the form of standardized experiment metadata about the specimen source analyzed and marker genes that serve as the most useful features in machine learning-based cell type classification models." @Aevermann2018 describe how the FAIR principles can be implemented using statistical identification of necessary and sufficient conditions for determining cell class membership. They propose that Cell Ontology can be transformed to a broadly usable knowledgebase through the incorporation of accurate marker gene signatures for cell classes.
In this vignette, we review key concepts and tasks required to make progress in the adoption and application of ontological discipline in Bioconductor-oriented data analysis.
We'll start by setting up some package attachments and ontology objects.
suppressPackageStartupMessages({ library(knitr) library(ontoProc) go = getGeneOnto() cl = getCellOnto() pr = getPROnto() library(ontologyPlot) library(BiocStyle) })
Scope of package
The following table describes the resources available with get*
commands defined in ontoProc.
kable(packDesc2019)
Methods
Conceptual overview of ontology with cell types
Definitions, semantics. For concreteness, we provide some definitions and examples.
We use ontology to denote the systematic organization
of terminology used in a conceptual domain. The
Cell Ontology is a graphical data structure with
carefully annotated terms as nodes and conventionally defined
semantic relationships among terms serving as edges. As
an example, lung ciliated cell has URI \url{http://purl.obolibrary.org/obo/CL_1000271}.
This URI includes a fixed-length identifier CL_1000271 with
unambiguous interpretation wherever it is encountered. There is
a chain of relationships from lung ciliated cell
up through ciliated cell, then native cell, then
cell, each possessing its own URI and related
interpretive metadata. The relationship connecting the more precise
to the less precise term in this chain
is denoted SubclassOf. Ciliated cell is equivalent to
a native cell that has plasma membrane part
cilium. Semantic characteristics of terms and relationships are
used to infer relationships among terms that may
not have relations directly specified in available ontologies.
Barriers to broad adoption. Given the wealth of material available in biological
ontologies, it is somewhat surprising that formal annotation
is so seldom used in practice. Barriers to
more common use of ontology in data annotation
include: (i) Non-existence of exact matching between intended
term and terms available in ontologies of interest.
(ii) The practical problem of decoding ontology identifiers.
A GO tag or CL tag is excellent
for programming, but it is clumsy to co-locate
with the tag the associated natural language term
or phrase. (iii) Likelihood of disagreement of suitability
of terms for conditions observed at the boundaries
of knowledge. To help cope with the first
of these problems, Bioconductor's ontologyProc package
includes a function liberalMap
which will search an ontology for terms lexically
close to some target term or phrase. The
second problem can be addressed with more elaborate
data structures for variable annotation and programming in
R, and the third problem will diminish in
importance as the value of ontology adoption becomes
manifest in more applications.
Class vs. instance. It is important to distinguish the practice of designing and maintaining ontologies from the use of ontological class terms to annotate instances of the concepts. The combination of an ontology and a set of annotated instances is called a knowledge base. To illustrate some of the salient distinctions here, consider the cell line called A549, which is established from a human lung adenocarcinoma sample. There is no mention of A549 in the Cell Ontology. However, A549 is present in the EBI Experimental Factor Ontology as a subclass of the "Homo sapiens cell line" class. Presumably this is because A549 is a class of cells that are widely used experimentally, and this cell line constitutes a concept deserving of mapping in the universe of experimental factors. In the universe of concepts related to cell structure and function per se, A549 is an individual that can be characterized through possession of or lack of properties enumerated in Cell Ontology, but it is not deserving of inclusion in that ontology.
Illustration in a single-cell RNA-seq dataset
The 10X Genomics corporation has distributed a dataset on results of sequencing 10000 PBMC from a healthy donor \url{https://support.10xgenomics.com/single-cell-gene-expression/datasets}. Subsets of the data are used in tutorials for the Seurat analytical suite (@butler).
Labeling PBMC in the Seurat tutorial
One result of the tutorial analysis of the 3000 cell subset is a table of cell types and expression-based markers of cell identity. The first three columns of the table below are from concluding material in the Seurat tutorial; the remaining columns are created by "manual" matching between the Seurat terms and terms found in Cell Ontology.
kable(stab <- seur3kTab())
Relationships asserted in the Cell Ontology
Given the informally selected tags in the table above, we can
sketch the Cell Ontology graph connecting the associated
cell types. The ontoProc package adds functionality to
ontologyPlot with make_graphNEL_from_ontology_plot. This
allows use of all Rgraphviz and igraph visualization facilities
for graphs derived from ontology structures.
Here we display the PBMC cell sets reported in the Seurat tutorial.
library(ontoProc) cl = getCellOnto() onto_plot2(cl, stab$tag)
Molecular features asserted in the Cell Ontology
The CLfeats function traces relationships and
properties from a given Cell Ontology class.
Briefly, each class can assert that it is the
intersection_of other classes, and
has_part, lacks_part, has_plasma_membrane_part,
lacks_plasma_membrane_part can be asserted as
relationships holding between cell type instances
and cell components. The components are often cross-referenced
to Protein Ontology or Gene Ontology. When the Protein Ontology
component has a synonym for which an HGNC symbol is provided, that
symbol is retrieved by CLfeats. Here we obtain the listing
for a mature CD1a-positive dermal dendritic cell.
kable(CLfeats(cl, "CL:0002531"))
The ctmarks function starts a shiny app that generates
tables of this sort for selected cell types.
Mapping from gene 'presence/role' to cell type
The sym2CellOnto function helps find mention of
given gene symbols in properties or parts of cell types.
kable(sdf <- as.data.frame(sym2CellOnto("ITGAM", cl, pr))) table(sdf$cond) kable(as.data.frame(sym2CellOnto("FOXP3", cl, pr)))
Adding terms to ontology_index structures to 'extend' Cell Ontology
The task of extending an ontology is partly bureaucratic in
nature and depends on a collection of endorsements and updates
to centralized information structures. In order to permit
experimentation with interfaces and new content that may
be quite speculative, we include an approach to combining new
ontology 'terms' of structure similar to those endorsed in
Cell Ontology, to ontologyIndex-based ontology_index
instances.
Use case: a set of cell types defined by "diagonal expression"
For a demonstration, we consider the discussion in @Bakken2017, of a 'diagonal' expression pattern defining a group of novel cell types. A set of genes is identified and cells are distinguised by expressing exactly one gene from the set.
The necessary information is collected in a vector. The vector is the set of genes, the name of element i is the tag to be associated with the type of cell that expresses gene i and does not express any other gene in the set.
sigels = c("CL:X01"="GRIK3", "CL:X02"="NTNG1", "CL:X03"="BAGE2", "CL:X04"="MC4R", "CL:X05"="PAX6", "CL:X06"="TSPAN12", "CL:X07"="hSHISA8", "CL:X08"="SNCG", "CL:X09"="ARHGEF28", "CL:X10"="EGF")
A data.frame defining the cell types and their properties
The cyclicSigset function produces a data.frame instance
connecting cell types with the genes expressed or unexpressed.
cs = cyclicSigset(sigels) dim(cs) cs[c(1:5,9:13),] table(cs$cond)
It is expected that a tabular layout like this will suffice to handle general situations of cell type definition.
Translating the data.frame elements to OBO Term instances
The most complicated aspect of novel OBO term construction is the proper specifications of relationships with existing ontology components. A prolog that is mostly shared by all terms is generated programmatically for the diagonal pattern task.
makeIntnProlog = function(id, ...) { # make type-specific prologs as key-value pairs c( sprintf("id: %s", id), sprintf("name: %s-expressing cortical layer 1 interneuron, human", ...), sprintf("def: '%s-expressing cortical layer 1 interneuron, human described via RNA-seq observations' [PMID 29322913]", ...), "is_a: CL:0000099 ! interneuron", "intersection_of: CL:0000099 ! interneuron") }
The ldfToTerms API uses this to create a set of strings that can be parsed
as a term.
pmap = c("hasExp"="has_expression_of", lacksExp="lacks_expression_of") head(unlist(tms <- ldfToTerms(cs, pmap, sigels, makeIntnProlog)), 20)
The content in tms can then be appended to the content of the Cell Ontology cl.obo as
text for import with ontologyIndex::get_OBO.
Subsetting SingleR resources using ontological mapping
A data.frame mapping from informal to formal terms
Aaron Lun has produced a mapping from informal terms used in the Human Primary Cell Atlas to Cell Ontology tags. We provisionally include a copy of this mapping in ontoProc:
hpca_map = read.csv(system.file("extdata/hpca.csv", package="ontoProc"), strings=FALSE) head(hpca_map)
We will rename columns of this map for convenience of our bind_formal_tags method.
names(hpca_map) = c("informal", "formal") # obligatory for now
Binding formal tags to the HPCA data
I am turning this code off for now because there is no standard approach to getting the mapping from the SummarizedExperment yet. When SingleR merges the 'standardized' branch, this will come back.
Let's retrieve the HPCA data from SingleR:
library(SingleCellExperiment) library(celldex) hpca_sce = HumanPrimaryCellAtlasData()
Now bind the formal tags:
hpca_sce = bind_formal_tags(hpca_sce, "label.fine", hpca_map) length(unique(hpca_sce$label.ont))
We don't check for failed mappings:
length(xx <- which(is.na(hpca_sce$label.ont))) if (length(xx)>0) print(colData(hpca_sce)[xx,]) sum(hpca_sce$label.ont == "", na.rm=TRUE) # iPS and BM
Subsetting using the class hierarchy of Cell Ontology
cell_onto = ontoProc::getCellOnto() hpca_mono = subset_descendants( hpca_sce, cell_onto, "^monocyte$" ) table(hpca_mono$label.fine) table(hpca_mono$label.ont) # not much diversity hpca_tcell = subset_descendants( hpca_sce, cell_onto, "^T cell$" ) table(hpca_tcell$label.fine) table(hpca_tcell$label.ont) # uu = unique(hpca_tcell$label.ont) onto_plot2(cell_onto, uu)
Related tools
Inference on the identities of cells assayed in
a single cell transcriptomics experiment can be performed
using the Bioconductor celaref package. This package includes
a number of reference data resources providing whole-transcriptome
profiles of cells of known and unknown type.
An approach to systematically structuring data on cell-type
signatures, and conducting inference on cell types in
new experiments, is provided in the Hancock package,
under development.
A CRAN package that is very useful for
R programming with ontologies is ontologyIndex @Westbury2015. This
provides easily used functions for parsing ontologies in
the OBO format and for performing basic queries
on text fields and list structures.
References
Try the ontoProc package in your browser
Any scripts or data that you put into this service are public.
R Package Documentation
Browse R Packages
We want your feedback!
Note that we can't provide technical support on individual packages. You should contact the package authors for that.
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