Project description:Single nuclei RNAseq, in situ hybridization, and immunohistochemistry were used to study the cells comprising the mouse iris in its different functional states.
Project description:Tuberculosis Immune Reconstitution Inflammatory Syndrome (TB-IRIS) frequently complicates combined anti-retroviral therapy (ART) and anti-tubercular therapy in HIV-1 co-infected tuberculosis (TB) patients. The immunopathological mechanism underlying TB-IRIS is incompletely defined. Differential transcript abundance in PBMC from IRIS and control patients stimulated with heat killed H37Rv was determined by microarray Blood samples were collected during longitudinal observational studies of TB-IRIS patients and controls (both groups HIV-infected patients placed on antiretroviral treatment). PBMC were stimulated with heat killed H37Rv and RNA extracted.
Project description:Patients with HIV-associated TB are known to experience systemic hyperinflammation, clinically known as immune reconstitution inflammatory syndrome (IRIS), following the commencement of antiretroviral therapy (ART). No prognostic markers or biomarkers have been identified to date and little is known about the mechanism mediating the hyperinflammation. We recruited a prospective cohort of 63 patients with HIV-associated TB, 33 of whom developed TB-IRIS. Of which transcriptomic profiling was performed using longitudinal whole blood RNA samples from 15 non-IRIS and 17 TB-IRIS patients. Transcriptomic signatures that distinguish patients who would eventually develop IRIS were identified as early as week 0.5 (2-5 days post-ART) and predicted a downstream activation of proinflammatory cytokines. At the peak of IRIS (week 2), transcriptomic signatures were overrepresented by innate receptor signaling pathways including toll-like receptor, IL-1 receptor and TREM-1.
Project description:Previous Genome-wide association studies (GWASs) have identified susceptibility loci of primary angle closure glaucoma (PACG), but applicating these findings is difficult. Although shallow anterior chamber depth (ACD) and short axial length (AL) are characteristic phenotypes of PACG, current GWAS variants do not show any correlation with these features, calling for the identification of more risk factors . Here, thicker and enlarged iris was identified as a significant independent risk factors. By employing allelic-specific STARR-seq and assay for transposase-accessible chromatin using sequencing (ATAC-seq), we screened disease-related variants in linkage disequilibrium and identified 2 causative variants in iris. Alterations in these variants may contribute to the pathogenic iris phenotype by upregulating the expression of PLEKHA7 and C10orf53, potentially regulating PACG development and progression. Our efforts nominate the important role of iris and identify pathogenic SNP-target gene interactions for PACG, providing a potentially powerful approach for interpreting noncoding variation of diseases.
Project description:Tuberculosis Immune Reconstitution Inflammatory Syndrome (TB-IRIS) frequently complicates combined anti-retroviral therapy (ART) and anti-tubercular therapy in HIV-1 co-infected tuberculosis (TB) patients. The immunopathological mechanism underlying TB-IRIS is incompletely defined. Differential transcript abundance in PBMC from IRIS and control patients stimulated with heat killed H37Rv was determined by microarray
Project description:Tuberculosis-associated Immune Reconstitution Inflammatory Syndrome (TB-IRIS) is a common complication in HIV-TB coinfected patients receiving combined antiretroviral therapy (cART). While monocytes/macrophages play major roles in both HIV- and TB-infection individually, a putative contribution of monocytes to the development of TB-IRIS remains unexamined. To investigate the possible functional contribution of monocytes to TB-IRIS pathogenesis, one of our first steps was to apply a genome-wide microarray analysis in monocytes of HIV-TB co-infected patients shortly after cART initiation. Based on the coparison of gene profiles between the TB-IRIS group and the control group, the modulated genes and pathways will be further investigated.
Project description:The human iris tissue is a thin, circular structure in the eye and it is made up of a pigmented epithelial structure. It is a protected internal organ of the eye, located behind the cornea and the aqueous humour. Iris serves main function to control the diameter, size of the pupil and regulation of light exposure to the internal eye structures. Damage or absent iris always results in allowing excess amount of light into the eye which causes medical problem for the patient and also a psychological problem due to strange eye with black hole. A damaged or congenitally defective iris does not function well which results in poor quality of vision. Although different efforts have been made to elucidate the different parts of the human eye proteome in depth, the protein composition of the human iris tissue remains largely unexplored. We have performed a comprehensive analysis of the human iris tissue employing protein and peptide fractionation methods followed by LC-MS/MS identifying 4918 proteins. Bioinformatics analysis revealed that protein components of the iris tissue participated in a plethora of biological process highlighting cell signal transduction, communication, metabolism, energy pathways protein metabolism cell growth and maintenance, transport and immune response activities. We also compared the proteins of iris tissue with high throughput studies on other parts of eye and plasma proteome, which resulted in identifying proteins unique to iris. To our knowledge, this study is the first attempt to profile the global proteome of the human iris tissue. Taken together, these results increase our knowledge about the molecular composition of the human iris tissue and may be useful to understand the molecular basis of the iris and the baseline proteome described in this study should serve as a resource for future research in iris tissue
Project description:Patients with HIV-associated TB meningitis (TBM) are known to experience systemic hyperinflammation, clinically known as immune reconstitution inflammatory syndrome (IRIS), following the commencement of antiretroviral therapy (ART). No prognostic markers or biomarkers have been identified to date and little is known about the mechanism mediating the hyperinflammation. We recruited a prospective cohort of 33 patients with HIV-associated TBM, 16 of whom developed TBM-IRIS, and performed transcriptomic profiling. Transcriptomic signatures that distinguish patients who would eventually develop IRIS were identified and indicated neutrophil and inflammasome activation as being the predominant mediator of TBM-IRIS pathogenesis.
Project description:The iris is a fine structure that controls the amount of light that enters the eye. The ciliary body controls the shape of the lens and produces aqueous humor. The retinal pigment epithelium and choroid (RPE/choroid) are essential in supporting the retina and absorbing light energy that enters the eye. Proteins were extracted from iris, ciliary body, and RPE/choroid tissues of eyes from five individuals and fractionated using SDS-PAGE. After in-gel digestion, peptides were analyzed using LC-MS/MS on an Orbitrap Elite mass spectrometer. In iris, ciliary body, and RPE/choroid, we identified 2,959, 2,867, and 2,755 non-redundant proteins with protein false positive rate <1%. There were 43 unambiguous protein isoforms identified in iris, ciliary body, and RPE/choroid. Four “missing proteins” were found in ciliary body. The MS proteome database of the human iris, ciliary body, and RPE/choroid may serve as a valuable resource for future investigations of the eye in health and disease. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD001424.