Project description:PRM in positive mode of baboon blood sample infected with T. Cruzi using polar-C18 columns in 12.5 minute run on a Q Exactive Plus.
Project description:5-week old male and female Swiss Webster mice were infected with 500,000 Trypanosoma cruzi strain Sylvio X/104 parasites. Urine was collected from uninfected mice pre-infection, from infected and uninfected mice every week for 5 weeks post infection, from infected and uninfected mice 67 (females)/68 (males) days post infection, and from infected and uninfected mice 117 (females)/124 (males) days post infection for acute, mid-chronic, and chronic time-points. Metabolites were extracted with methanol and a targeted PRM LCMS analysis was ran on a Thermo Q Exactive Plus instrument with a Phenomenex Luna Omega Polar C18 column.
Project description:5-week old male and female Swiss Webster mice were infected with 500,000 Trypanosoma cruzi strain Sylvio X10/4 parasites. Urine was collected from uninfected mice pre-infection, from infected and uninfected mice every week for 5 weeks post infection, from infected and uninfected mice 67 (females)/68 (males) days post infection, and from infected and uninfected mice 117 (females)/124 (males) days post infection for acute, mid-chronic, and chronic time-points. Metabolites were extracted with methanol and an untargeted LCMS analysis was ran on a Thermo Q Exactive Plus instrument with a Phenomenex Luna Omega Polar C18 column.
Project description:5-week old male and female Swiss Webster mice were infected with 500,000 Trypanosoma cruzi strain Sylvio X/104 parasites. Urine was collected from uninfected mice pre-infection, from infected and uninfected mice every week for 5 weeks post infection, from infected and uninfected mice 67 (females)/68 (males) days post infection, and from infected and uninfected mice 117 (females)/124 (males) days post infection for acute, mid-chronic, and chronic time-points. Metabolites were extracted with methanol and a targeted PRM LCMS analysis was ran on a Thermo Q Exactive Plus instrument with a Phenomenex Luna Omega Polar C18 column.
Project description:Trypanosoma cruzi (T. cruzi), the causative agent of Chagas disease, remains a major global health threat lacking effective therapies. Macrophages are key effector cells in the host response to T. cruzi infection, yet their incomplete clearance of T. cruzi contributes to Chagas disease pathogenesis. 5’ isomiRs, miRNA isoforms with shifted 5’ ends and altered seed sequences, have emerged as important regulators in diverse diseases. However, their expression alternations in T. cruzi-infected macrophages remain largely unknown. Here, we performed comparative analyses of 5’ isomiR expression in T. cruzi-infected macrophages. We found that T. cruzi infection induced a pronounced and selective increase in miRNA 5’-end heterogeneity in THP-1-derived macrophages, but not in cardiomyocytes or epithelial cells. Comparative 5’ isomiRome analyses further identified 68 differentially expressed 5’ isomiRs in THP-1 macrophages, most of which were specific to T. cruzi infection compared with other pathogens. Among these, 56 5’ isomiRs were associated with Argonaute proteins, including three originating from the miR-1246 precursor. Focusing on miR-1246|+1, a 5’ isomiR generated by a one-nucleotide downstream shift at the 5’ end, we showed that its overexpression significantly downregulated target genes involved in NF-κB signaling, cytokine responses, and cell migration. Notably, miR-1246|+1 broadly suppressed NF-κB family transcription factors and their downstream effector genes, thereby inhibiting pro-inflammatory M1 macrophage phenotype. Collectively, these findings reveal cell type- and pathogen-specific reprogramming of the 5’ isomiR landscape in T. cruzi-infected macrophages and identify miR-1246|+1 as a potential post-transcriptional regulator of macrophage inflammatory polarization.
Project description:As Trypanosoma cruzi, the etiological agent of Chagas disease, multiplies in the cytoplasm of nucleated host cells, infection with this parasite is highly likely to affect host cells. We performed an exhaustive transcriptome analysis of T. cruzi-infected HeLa cells using an oligonucleotide microarray containing probes for greater than 47,000 human gene transcripts. In comparison with uninfected cells, those infected with T. cruzi showed greater than threefold up-regulation of 41 genes and greater than threefold down-regulation of 23 genes. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) of selected, differentially expressed genes confirmed the microarray data. Many of these up- and down-regulated genes were related to cellular proliferation, including seven up-regulated genes encoding proliferation inhibitors and three down-regulated genes encoding proliferation promoters, strongly suggesting that T. cruzi infection inhibits host cell proliferation, which may allow more time for T. cruzi to replicate and produce its intracellular nests. These findings provide new insight into the molecular mechanisms by which intracellular T. cruzi infection influences the host cell, leading to pathogenicity. Keywords: infection response
Project description:In this study, we conducted integrative high-throughput analyses of peripheral blood and placental tissue from naturally infected patients in Bolivia, the country with the highest prevalence of Chagas disease and, consequently, the population with the greatest potential for congenital transmission worldwide. We compared T. cruzi-infected and uninfected patients to characterize parasite-associated alterations. Infected mothers were further classified as transmitters or non-transmitters based on the infant’s infection status, determined by serological and molecular tests. Transmitter and non-transmitter mothers were compared to understand the molecular mechanism associated with congenital transmission risk.
Project description:Investigate transcriptional responses of T. cruzi infected hiPSC-derived cardiomyocytes from chagasic patients with and without cardiomyopathy.