22,572 research outputs found
Regulatory T cells in melanoma revisited by a computational clustering of FOXP3+ T cell subpopulations
CD4+ T cells that express the transcription factor FOXP3 (FOXP3+ T cells) are commonly regarded as immunosuppressive regulatory T cells (Treg). FOXP3+ T cells are reported to be increased in tumour-bearing patients or animals, and considered to suppress anti-tumour immunity, but the evidence is often contradictory. In addition, accumulating evidence indicates that FOXP3 is induced by antigenic stimulation, and that some non-Treg FOXP3+ T cells, especially memory-phenotype FOXP3low cells, produce proinflammatory cytokines. Accordingly, the subclassification of FOXP3+ T cells is fundamental for revealing the significance of FOXP3+ T cells in tumour immunity, but the arbitrariness and complexity of manual gating have complicated the issue. Here we report a computational method to automatically identify and classify FOXP3+ T cells into subsets using clustering algorithms. By analysing flow cytometric data of melanoma patients, the proposed method showed that the FOXP3+ subpopulation that had relatively high FOXP3, CD45RO, and CD25 expressions was increased in melanoma patients, whereas manual gating did not produce significant results on the FOXP3+ subpopulations. Interestingly, the computationally-identified FOXP3+ subpopulation included not only classical FOXP3high Treg but also memory-phenotype FOXP3low cells by manual gating. Furthermore, the proposed method successfully analysed an independent dataset, showing that the same FOXP3+ subpopulation was increased in melanoma patients, validating the method. Collectively, the proposed method successfully captured an important feature of melanoma without relying on the existing criteria of FOXP3+ T cells, revealing a hidden association between the T cell profile and melanoma, and providing new insights into FOXP3+ T cells and Treg
Essential guidelines for computational method benchmarking
In computational biology and other sciences, researchers are frequently faced
with a choice between several computational methods for performing data
analyses. Benchmarking studies aim to rigorously compare the performance of
different methods using well-characterized benchmark datasets, to determine the
strengths of each method or to provide recommendations regarding suitable
choices of methods for an analysis. However, benchmarking studies must be
carefully designed and implemented to provide accurate, unbiased, and
informative results. Here, we summarize key practical guidelines and
recommendations for performing high-quality benchmarking analyses, based on our
experiences in computational biology.Comment: Minor update
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Mapping lung cancer epithelial-mesenchymal transition states and trajectories with single-cell resolution.
Elucidating the spectrum of epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) states in clinical samples promises insights on cancer progression and drug resistance. Using mass cytometry time-course analysis, we resolve lung cancer EMT states through TGFβ-treatment and identify, through TGFβ-withdrawal, a distinct MET state. We demonstrate significant differences between EMT and MET trajectories using a computational tool (TRACER) for reconstructing trajectories between cell states. In addition, we construct a lung cancer reference map of EMT and MET states referred to as the EMT-MET PHENOtypic STAte MaP (PHENOSTAMP). Using a neural net algorithm, we project clinical samples onto the EMT-MET PHENOSTAMP to characterize their phenotypic profile with single-cell resolution in terms of our in vitro EMT-MET analysis. In summary, we provide a framework to phenotypically characterize clinical samples in the context of in vitro EMT-MET findings which could help assess clinical relevance of EMT in cancer in future studies
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Integrated single cell analysis of blood and cerebrospinal fluid leukocytes in multiple sclerosis
Essential guidelines for computational method benchmarking
In computational biology and other sciences, researchers are frequently faced with a choice between several computational methods for performing data analyses. Benchmarking studies aim to rigorously compare the performance of different methods using well-characterized benchmark datasets, to determine the strengths of each method or to provide recommendations regarding suitable choices of methods for an analysis. However, benchmarking studies must be carefully designed and implemented to provide accurate, unbiased, and informative results. Here, we summarize key practical guidelines and recommendations for performing high-quality benchmarking analyses, based on our experiences in computational biology
Joint Modeling and Registration of Cell Populations in Cohorts of High-Dimensional Flow Cytometric Data
In systems biomedicine, an experimenter encounters different potential
sources of variation in data such as individual samples, multiple experimental
conditions, and multi-variable network-level responses. In multiparametric
cytometry, which is often used for analyzing patient samples, such issues are
critical. While computational methods can identify cell populations in
individual samples, without the ability to automatically match them across
samples, it is difficult to compare and characterize the populations in typical
experiments, such as those responding to various stimulations or distinctive of
particular patients or time-points, especially when there are many samples.
Joint Clustering and Matching (JCM) is a multi-level framework for simultaneous
modeling and registration of populations across a cohort. JCM models every
population with a robust multivariate probability distribution. Simultaneously,
JCM fits a random-effects model to construct an overall batch template -- used
for registering populations across samples, and classifying new samples. By
tackling systems-level variation, JCM supports practical biomedical
applications involving large cohorts
Cluster analysis of flow cytometric list mode data on a personal computer
A cluster analysis algorithm, dedicated to analysis of flow cytometric data is described. The algorithm is written in Pascal and implemented on an MS-DOS personal computer. It uses k-means, initialized with a large number of seed points, followed by a modified nearest neighbor technique to reduce the large number of subclusters. Thus we combine the advantage of the k-means (speed) with that of the nearest neighbor technique (accuracy). In order to achieve a rapid analysis, no complex data transformations such as principal components analysis were used. \ud
Results of the cluster analysis on both real and artificial flow cytometric data are presented and discussed. The results show that it is possible to get very good cluster analysis partitions, which compare favorably with manually gated analysis in both time and in reliability, using a personal computer
Identifying Mixtures of Mixtures Using Bayesian Estimation
The use of a finite mixture of normal distributions in model-based clustering
allows to capture non-Gaussian data clusters. However, identifying the clusters
from the normal components is challenging and in general either achieved by
imposing constraints on the model or by using post-processing procedures.
Within the Bayesian framework we propose a different approach based on sparse
finite mixtures to achieve identifiability. We specify a hierarchical prior
where the hyperparameters are carefully selected such that they are reflective
of the cluster structure aimed at. In addition this prior allows to estimate
the model using standard MCMC sampling methods. In combination with a
post-processing approach which resolves the label switching issue and results
in an identified model, our approach allows to simultaneously (1) determine the
number of clusters, (2) flexibly approximate the cluster distributions in a
semi-parametric way using finite mixtures of normals and (3) identify
cluster-specific parameters and classify observations. The proposed approach is
illustrated in two simulation studies and on benchmark data sets.Comment: 49 page
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