Computational Tools for Stem Cell Biology

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Computational Tools for Stem Cell Biology Qin Bian, Patrick Cahan  Trends in Biotechnology  Volume 34, Issue 12, Pages 993-1009 (December 2016) DOI: 10.1016/j.tibtech.2016.05.010 Copyright © 2016 Elsevier Ltd Terms and Conditions

Figure 1 Key Figure: Computational Tools to Address the Three Major Barriers to Achieving Cell Fate Engineering For a Figure360 author presentation of Figure 1, see the figure online at http://dx.doi.org/10.1016/j.tibtech.2016.05.010#mmc1. (A) The ‘Improvement problem’: how can we devise protocols to improve the fidelity of engineered cells? (B) The ‘Assessment problem’: to what extent is the engineered population equivalent to the desired cell type? (C) The ‘Lineage bias problem’: how can we quantify the in vitro lineage bias of a pluripotent stem cell (PSC) line? Figure360: an author presentation of Figure 1 Trends in Biotechnology 2016 34, 993-1009DOI: (10.1016/j.tibtech.2016.05.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Figure 2 Single Cell Molecular Profiling to Identify Rare Populations, to Reconstruct Gene Regulatory Networks, and to Define Lineage Trajectories. (A–C) Distinguishing expression heterogeneity from population substructure. (A) An example of applying dimension reduction, in this case principal component analysis (PCA), to single cell RNA-Seq data comprising two major cell types (a and b). The analytical task is to determine whether two (red and light blue) single cells should be considered as distinct, rare cell types or are more likely to represent statistical outliers. (B) A background model of transcriptional noise (based on the average transcript count variance across all cells) is used to weigh the relative likelihood that the outlier cells belong to one of the predetermined clusters or represent a distinct class. (C) Identification of distinct type of cells (a′ and b′). (D,E) Gene regulatory network (GRN) reconstruction. Reconstructing GRNs from expression profiling (either array or RNA-Seq) is based on correlations in expression between genes. Expression profiling from bulk samples can stymie this type of analysis by producing both false positives and false negatives. (D) False positives can occur when a correlation is a result of population substructure rather than because of a regulatory relationship. (E) Similarly, population substructure can mask regulatory relationships present on subpopulation. (F–K) Inferring lineage trajectory. Steps common to lineage trajectory inference algorithms include: (F) reducing the dimensionality of the data (every point represents a single cell expression profile), and (G) finding a minimal spanning tree. Methods differ in how they assign a path through the minimal spanning tree to lineage progression, with Monocle using the longest path (H) and Wanderlust using an average of all possible paths (I). (J,K) The resulting path is referred to as pseudotime and can be used to order cells in a temporal progression. Trends in Biotechnology 2016 34, 993-1009DOI: (10.1016/j.tibtech.2016.05.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Figure I A Schematic of Seven Analytical Tools Commonly Applied to OMICs Data. (A) Hierarchical clustering (HCL) samples based on their similarity. Panels (A–F) represent different samples. (B) K-means divides variables into a user-selected number of groups. (C) Principal component analysis (PCA) reduces the number of dimensions in data. (D) Two duplicates of each condition. Gene 1 is considered differentially expressed, whereas genes 2 and 3 are not. (E) GSEA showing whether a set of genes is statistically significantly different between two conditions. (F) A→G mutation detected by next generation sequencing (NGS). (G) Peak comparison between two conditions. Trends in Biotechnology 2016 34, 993-1009DOI: (10.1016/j.tibtech.2016.05.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Figure I A Schematic of Three Machine Learning Classifiers. (A) Support vector machines (SVMs) aim to separate two groups (green and red). (B) Given expression distributions of gene x in each of red blood cells (RBCs) and hematopoietic stem cells (HSCs), it is possible to compute the posterior probability that a sample is either a RBC or HSC using Bayes theorem (Naïve Bayes classifier, NBC). For convenience, the prior probability is usually assumed to be equal across all cell types. (C) A random forest (RF) classifier treats the classification of each decision tree as a vote and returns the classification with the greatest number of votes. Trends in Biotechnology 2016 34, 993-1009DOI: (10.1016/j.tibtech.2016.05.010) Copyright © 2016 Elsevier Ltd Terms and Conditions