Project description:We compared multiple strains of lab trophozoites to recent clinical isolates. Clinical isolates were grown in xenic media, and maintained many characteristics of the cyst stage of devlopment Keywords: Stage conversion
Project description:Pseudomonas aeruginosa (P. aeruginosa) lung infection is a significant cause of mortality in patients with cystic fibrosis (CF). Existing experimental data in our lab showed significantly different levels of virulence of "early" and "late" P. aeruginosa infection isolates in a C. elegans slow killing model. We wished to examine the expression profile of these isolates in order to explore genes that may be responsible for the observed differences. The expression profiles of two pairs of isolates (four isolates in total) were compared to each other using the Affymetrix P. aeruginosa PAO1 genome array, to gain insight into properties mediating virulence in these isolates. Data analysis was carried out using BIOCONDUCTOR software. Keywords: Comparative strain hybridization
Project description:Atherosclerosis is the main underlying cause of cardiovascular diseases (CVDs). Its prevention is based on traditional cardiovascular risk factor-based scores but often fails to identify individuals at early stages of the disease. Here, we identified microbially produced imidazole propionate (ImP) as an early biomarker of atherosclerosis in mice and in two independent human cohorts. Furthermore, ImP administration induced atherosclerosis without altering lipid metabolism and it was associated with activation of both systemic and local innate and adaptive immunity and inflammation. Here, we used single-cell RNA-seq to characterize the local changes of aorta-derived cells in mice under treatment with ImP for 4 and 8 weeks. Notably, ImP caused atherosclerosis through the Imidazoline-1 receptor (I1R), and the blockade of the ImP/I1R axis inhibited the development of atherosclerosis induced by ImP as well as by high cholesterol diet in mice. Identification of ImP as an early biomarker for atherosclerosis and uncovering the contribution of the ImP/I1R axis in disease progression open new avenues to improve atherosclerosis early diagnosis and therapy.
Project description:Atherosclerosis is the main underlying cause of cardiovascular diseases (CVDs). Its prevention is based on traditional cardiovascular risk factor-based scores but often fails to identify individuals at early stages of the disease. Here, we identified microbially produced imidazole propionate (ImP) as an early biomarker of atherosclerosis in mice and in two independent human cohorts. Furthermore, ImP administration induced atherosclerosis without altering lipid metabolism and it was associated with activation of both systemic and local innate and adaptive immunity and inflammation. Notably, ImP caused atherosclerosis through the Imidazoline-1 receptor (I1R), and the blockade of the ImP/I1R axis inhibited the development of atherosclerosis induced by ImP as well as by high cholesterol diet in mice. Here, we profiled three cell lines which represent cell types typically found in aortas (bone marrow-derived macrophages, mouse embryonic fibroblasts and mouse aortic endothelial cells) upon in vitro stimulation with ImP at different time points to better understand the transcriptional regulation of the ImP/I1R axis and its role in the development of the disease. Identification of ImP as an early biomarker for atherosclerosis and uncovering the contribution of the ImP/I1R axis in disease progression open new avenues to improve atherosclerosis early diagnosis and therapy.
Project description:Neural circuits driving mammalian behaviors are highly plastic and modulated by internal and external factors, including the gut microbiome. We identify imidazole propionate (ImP), a microbial metabolite linked to metabolic disorders, as a key modulator of brain activity and behavior. Bacterially derived ImP enters the systemic circulation and brain, where it alters neuronal gene expression and activity in the hypothalamus without inducing overt neuroinflammation. Elevating systemic ImP promotes stress-related pathways and disrupts GABAergic/glutamatergic signaling in the hypothalamus before peripheral glucose dysregulation occurs. Similarly, colonization with the ImP-producing bacterium Eggerthella lenta elevates behavioral and molecular features of stress. In a mouse model of type 2 diabetes, the gut microbiome exhibits an enhanced ability to produce ImP, leading to elevated systemic levels that are associated with heightened stress responses. In humans, higher ImP associates with reduced hypothalamic reactivity to food cues, impaired stress-coping, and increased emotional eating, mirroring the causal links between ImP, hypothalamic activity, and stress-related behaviors in mice. Overall, these findings establish ImP as a notable microbial metabolite that links gut dysbiosis to altered hypothalamic function and stress in metabolic disease.
Project description:Recent advancements in genome sequencing have facilitated accessing the natural genetic diversity of species, unveiling hidden genetic traits, clarifying gene functions, and the degree to which laboratory studies can be generalized. One notable discovery is the frequent (~20%) aneuploidy - an imbalance in chromosome copy numbers - in natural Saccharomyces cerevisiae (Sc) isolates, despite the significant fitness costs and transient nature reported for lab-engineered yeast aneuploids. To examine this discrepancy, we adapted a high-throughput proteomic platform to analyze the proteome of 800 diverse yeast isolates. Matching these proteomes to the natural isolates’ genomes, transcriptomes, as well as generating ubiquitinome and protein turnover data for selected isolates, we report that natural and lab-generated aneuploids differ specifically at the proteome. While lab-generated aneuploids attenuate specific proteins – mostly protein complex subunits – and do not alter the average gene dosage provided by chromosome duplications, in natural strains, 70% of proteins encoded on aneuploid chromosomes are attenuated, and protein levels are shifted towards the euploid state chromosome-wide. Our data links chromosome-wide dosage compensation in natural strains to i) genome-wide buffering of gene expression changes manifesting in trans on euploid chromosomes, ii) increased expression of structural components of the ubiquitin proteasome system, and iii) increased global rates of protein turnover. Our results encourage the exploitation of natural diversity of species to understand complex biological processes at the molecular level. This submission contains the raw files for the disomics lab engineered strains, the library used for the analysis and the corresponding DIA-NN report and associated files.
Project description:Inosine monophosphate (IMP) plays a central role in intracellular purine metabolism. How IMP regulates inflammation induced by bacterial products or bacteria is unknown. In this study, we demonstrate that IMP fundamentally alters gene expression in the liver.To assess how IMP affects inflammatory cell activation in vivo, we subjected liver, the organ containing the highest numbers of macrophages in the body, to RNAseq analyses. High quality RNA was isolated from the livers of 3 vehicle- and 3 IMP-treated endotoxemic mice and submitted for RNAseq analysis. Our analysis identified 105 genes that were differentially expressed, determined by adjusted p values (42 downregulated, 63 upregulated; padj ≤ 0.05).
2021-09-12 | GSE183880 | GEO
Project description:clinical IMP-producing Klebsiella spp. Isolates in China
Project description:To identify factors and pathways regulated by IMP proteins and obtain leads to the mechanism behind the phenotypic changes, we compared the gene expression profiles of IMP siRNA treated cells with mock treated cells. Triplicate gene expression profiles were generated from both the IMP(1,3)A and IMP(1,3)B siRNA sets and were compared to the mock transfected cells. cRNA was hybridized to Affymetrix human U133A arrays.