Project description:This study represents the first in vivo genome wide analysis of gene epxression of Group A Streptococcus (GAS) in humans with pharyngitis. The micorarray used is a custom microarray that relies on an electrochemical reaction as the measured signal rather than flourescence. Distinct clusters of gene expression were discovered and analyzed. A functional analysis examining differences and similarities between the clusters was performed. Samples were taken from pediatric patients who had received a throat swab as part of their clinical care for evaluating pharyngitis. Eleven samples that were culture postive for GAS were used along with 3 samples from subjects who were GAS culture negative. The microarray was a composite comprised of 12,000 probes, representing 2724 GAS open reading frames belonging to serotypes M1, M3, M4, M12, and M28. There were 1671 probe sets that were homologous for the M1 serotype and represented over 95% of the total predicted coding region. Probe sets were 30-40mers in length and were selected using a Combimatrix probe selection algorithm taking into account gene (>90% BLAST score) and serotype specificity, Tm, hairpins and GC content. In addition, 70 Combimatrix built in probes were used as negative controls for background subtraction.
Project description:Better understanding of S. aureus throat colonization in the presence of other competing/coexisting microbes may provide insight into S. aureus adaptation to the human throat and recurrence of infection. In this work, we explore the responses triggered by the encounter between two common throat bacteria, S. aureus and S. anginosus, in the presence of human tonsillar epithelial cells. We performed an in vitro coculture experiment followed by RNA sequencing. A total of 332 and 279 significant DEGs with p-value < 0.05 and Log2 fold change > |2| were identified after 1 h and 3 h of post-infection, respectively. Our study identified several transcripts in S. aureus that might be important when facing a potential competitor during throat colonization. These transcripts may be useful in the development of treatments against S. aureus throat colonization and could be further investigated to better understand their roles in the immune response.
Project description:We compared PPARg binding sites in BAT and eWAT to identify regulatory elements that contribute to BAT identity and to find an important factor that bind those elements. To this end, we performed PPARg ChIP-seq in both tissues and called each tissue-spsecific binding sites. PPARg ChIP-seq in BAT and eWAT of mice
Project description:Nasal colonization is well described; however, we have limited knowledge about S. aureus throat colonization. The main objective of this project was to explore differentially expressed genes (DEGs) in S. aureus throat isolate TR145 exposed for 1 or 3 hours (h) to human tonsil epithelial cells (HTEpiC) by using RNA sequencing (RNA-seq) and pathway analysis. We have shown the suitability of using HTEpiC as an in vitro model for investigating key determinants in S. aureus during co-incubation with the HTEpiC cells. Among the DEGs were genes encoding proteins involved in adhesion and immune evasion, as well as iron acquisition and transport. As their expression is induced upon meeting with the HTEpiC, they might be explored further for future targets for intervention to prevent either colonization or infection in the throat region.
Project description:Brown adipose tissue (BAT) is a thermogenic organ that protects animals against hypothermia and obesity. BAT derives from the multipotent paraxial mesoderm; however, the identity of embryonic brown fat progenitor cells and regulators of adipogenic commitment are unclear. We identified the transcription factor GATA6 as a selective marker of brown adipogenic progenitor cells. Deletion of Gata6 in the brown fat lineage resulted in a striking loss of BAT. To gain insight into the mechanism by which GATA6 supports BAT development, we performed ChIP-seq for GATA6 from the BAT of embryonic day 15.5 embryos.
Project description:Group A Streptococcus (GAS) has a rich evolutionary history of horizontal transfer among its core genes. Yet, despite extensive genetic mixing, GAS strains have discrete ecological phenotypes. To further our understanding of the molecular basis for ecological phenotypes, comparative genomic hybridization of a set of 97 diverse strains to a GAS pan-genome microarray was undertaken, and the association of accessory genes with emm genotypes that define tissue tropisms for infection was determined. Of the 22 non-prophage, accessory gene regions (AGRs) identified, only three AGRs account for all statistically significant linkage disequilibrium among strains having the genotypic biomarkers for throat versus skin infection specialist. Networked evolution and population structure analysis of loci representing each of the AGRs reveals that most strains with the skin specialist and generalist biomarkers form discrete clusters, whereas strains with the throat specialist biomarker are highly diverse. To identify co-inherited and co-selected accessory genes, the strength of genetic associations was determined for all possible pair wise combinations of accessory genes among the 97 GAS strains. Accessory genes showing very strong associations provide the basis for an evolutionary model, which reveals that a major transition between many throat and skin specialist haplotypes correlates with the gain or loss of genes encoding fibronectin-binding proteins. This study employs a novel synthesis of tools to help delineate the major genetic changes associated with key adaptive shifts in an extensively recombined bacterial species.