Variations in Structural MRI Quality Impact Measures of Brain Anatomy Relations with Age and Other Sociodemographic Variables, bioRxiv, 2019-03-20

AbstractIn-scanner head movements can introduce artifacts to MRI images and increase errors in brain-behavior studies. The magnitude of in-scanner head movements varies widely across developmental and clinical samples, making it increasingly difficult to parse out “true signal” from motion related noise. Yet, the quantification of structural imaging quality is typically limited to subjective visual assessments andor proxy measures of motion. It is, however, unknown how direct measures of image quality relate to developmental and behavioral variables, as well as measures of brain morphometrics. To begin to answer this question, we leverage a multi-site dataset of structural MRI images, which includes a range of children and adolescents with varying degrees of psychopathology. We first find that a composite of structural image quality relates to important developmental and behavioral variables (e.g., IQ; clinical diagnoses). Additionally, we demonstrate that even among T1-weighted images which pass visual inspection, variations in image quality impact volumetric derivations of regional gray matter. Image quality was associated with wide-spread variations in gray matter, including in portions of the frontal, parietal, and temporal lobes, as well as the cerebellum. Further, our image quality composite partially mediated the relationship between age and total gray matter volume, explaining 23% of this relationship. Collectively, the effects underscore the need for volumetric studies to model or mitigate the effect of image quality when investigating brain-behavior relations.

Evolutionary pathways to antibiotic resistance are dependent upon environmental structure and bacterial lifestyle, bioRxiv, 2019-03-19

AbstractBacterial populations vary in their stress tolerance and population structure depending upon whether growth occurs in well-mixed or structured environments. We hypothesized that evolution in biofilms would generate greater genetic diversity than well-mixed environments and lead to different pathways of antibiotic resistance. We used experimental evolution and whole genome sequencing to test how the biofilm lifestyle influenced the rate, genetic mechanisms, and pleiotropic effects of resistance to ciprofloxacin in Acinetobacter baumannii populations. Both evolutionary dynamics and the identities of mutations differed between lifestyle. Planktonic populations experienced selective sweeps of mutations including the primary topoisomerase drug targets, whereas biofilm-adapted populations acquired mutations in regulators of efflux pumps. An overall trade-off between fitness and resistance level emerged, wherein biofilm-adapted clones were less resistant than planktonic but more fit in the absence of drug. However, biofilm populations developed collateral sensitivity to cephalosporins, demonstrating the clinical relevance of lifestyle on the evolution of resistance.

Gene regulatory network reconstruction using single-cell RNA sequencing of barcoded genotypes in diverse environments, bioRxiv, 2019-03-19

AbstractUnderstanding how gene expression programs are controlled requires identifying regulatory relationships between transcription factors and target genes. Gene regulatory networks are typically constructed from gene expression data acquired following genetic perturbation or environmental stimulus. Single-cell RNA sequencing (scRNAseq) captures the gene expression state of thousands of individual cells in a single experiment, offering advantages in combinatorial experimental design, large numbers of independent measurements, and accessing the interaction between the cell cycle and environmental responses that is hidden by population-level analysis of gene expression. To leverage these advantages, we developed a method for transcriptionally barcoding gene deletion mutants and performing scRNAseq in budding yeast (Saccharomyces cerevisiae). We pooled diverse genotypes in 11 different environmental conditions and determined their expression state by sequencing 38,285 individual cells. We developed, and benchmarked, a framework for learning gene regulatory networks from scRNAseq data that incorporates multitask learning and constructed a global gene regulatory network comprising 12,018 interactions. Our study establishes a general approach to gene regulatory network reconstruction from scRNAseq data that can be employed in any organism.

Predicting the effects of SNPs on transcription factor binding affinity, bioRxiv, 2019-03-19

AbstractGWAS have revealed that 88% of disease associated SNPs reside in noncoding regions. However, noncoding SNPs remain understudied, partly because they are challenging to prioritize for experimental validation. To address this deficiency, we developed the SNP effect matrix pipeline (SEMpl). SEMpl estimates transcription factor binding affinity by observing differences in ChIP-seq signal intensity for SNPs within functional transcription factor binding sites genome-wide. By cataloging the effects of every possible mutation within the transcription factor binding site motif, SEMpl can predict the consequences of SNPs to transcription factor binding. This knowledge can be used to identify potential disease-causing regulatory loci.

Stag1 and Stag2 regulate cell fate decisions in hematopoiesis through non-redundant topological control, bioRxiv, 2019-03-19

AbstractTranscriptional regulators, including the cohesin complex member STAG2, are recurrently mutated in cancer. The role of STAG2 in gene regulation, hematopoiesis, and tumor suppression remains unresolved. We show Stag2 deletion in hematopoietic stemprogenitor cells (HSPC) results in altered hematopoietic function, increased self-renewal, and impaired differentiation. ChIP-sequencing revealed that while Stag2 and Stag1 can bind the same loci, a component of Stag2 binding sites are unoccupied by Stag1 even in Stag2-deficient HSPCs. While concurrent loss of Stag2 and Stag1 abrogated hematopoiesis, Stag2 loss alone decreased chromatin accessibility and transcription of lineage-specification genes, including Ebf1 and Pax5, leading to blunted HSPC commitment to the B-cell lineage. Our data illustrate a role for Stag2 in transformation and transcriptional dysregulation distinct from its shared role with Stag1 in chromosomal segregation.One Sentence SummaryStag1 rescues topologically associated domains in the absence of Stag2, but cannot restore chromatin architecture required for hematopoietic lineage commitment

The mesoSPIM initiative open-source light-sheet mesoscopes for imaging in cleared tissue, bioRxiv, 2019-03-19