Project description:Since microRNAs (miRNAs) have been associated with eye diseases, our study aims to profile ocular miRNA expression in normal human eye tissue using miRNA-Seq in order to provide a foundation for future disease research. Total RNAs were extracted from normal human ciliary body (CB) (n=7), cornea (n=7), and trabecular meshwork (TM) (n=7) samples using mirVana total RNA isolation kit. The sequencing library was prepared using the Illumina TruSeq Small RNA Sample Prep kit and was sequenced using Illumina MiSeq. Generated sequence reads were trimmed and aligned against human reference database using the Bowtie software, and only exact matches to mature miRNAs from miRBase were included. miRTarBase database was used to analyze the gene targets of the miRNAs in our tissues, and expression of a few selected miRNAs were validated using droplet digital PCR (ddPCR). We found 378 miRNAs expressed collectively in our samples, of which miR-143-3p, miR-184, miR-26a-5p, and miR-204-5p were most abundantly expressed. With the expression patterns and gene targets, we identified several uniquely expressed miRNAs and created a profile of miRNAs known to target genes associated with keratoconus and glaucoma. Using ddPCR, we were able to validate the expression profile created using miRNA-Seq. For the first time, we profiled miRNA expression in three human ocular tissues using miRNA-Seq, identifying many miRNAs that had not been previously reported in ocular tissue. Knowing the expression of miRNAs in non-diseased eye tissues could help elucidate miRNA changes that accompany diseases such as glaucoma and keratoconus.
Project description:Model animals are employed in experiments as substitutes for human tissues and fluids, particularly when accessing particular human samples (such as cerebrospinal fluid, brain, ocular tissues, etc.) poses significant challenges or is ethically constrained. Nonhuman primates are frequently regarded as superior animal models for investigating human ophthalmological diseases. However, despite this recognition, the metabolomic composition of ocular tissues in non-human primates remains predominantly unexplored. In this work, we present a dataset on metabolite concentrations in serum and ocular tissues, including aqueous humor (AH), vitreous humor (VH), and lens, in two Macaque species: crab-eating macaque (Macaca fascicularis) and rhesus macaque (Macaca mulatta). A total of 99 compounds were quantified in 45 samples, shedding light on the previously unknown metabolomic profiles of primate eye tissues.
Project description:Since microRNAs (miRNAs) have been associated with eye diseases, our study aims to profile ocular miRNA expression in normal human eye tissue using miRNA-Seq in order to provide a foundation for future disease research. Total RNAs were extracted from normal human ciliary body (CB) (n=7), cornea (n=7), and trabecular meshwork (TM) (n=7) samples using mirVana total RNA isolation kit. The sequencing library was prepared using the Illumina TruSeq Small RNA Sample Prep kit and was sequenced using Illumina MiSeq. Generated sequence reads were trimmed and aligned against human reference database using the Bowtie software, and only exact matches to mature miRNAs from miRBase were included. miRTarBase database was used to analyze the gene targets of the miRNAs in our tissues, and expression of a few selected miRNAs were validated using droplet digital PCR (ddPCR). We found 378 miRNAs expressed collectively in our samples, of which miR-143-3p, miR-184, miR-26a-5p, and miR-204-5p were most abundantly expressed. With the expression patterns and gene targets, we identified several uniquely expressed miRNAs and created a profile of miRNAs known to target genes associated with keratoconus and glaucoma. Using ddPCR, we were able to validate the expression profile created using miRNA-Seq. For the first time, we profiled miRNA expression in three human ocular tissues using miRNA-Seq, identifying many miRNAs that had not been previously reported in ocular tissue. Knowing the expression of miRNAs in non-diseased eye tissues could help elucidate miRNA changes that accompany diseases such as glaucoma and keratoconus. This study profiled the expression of miRNAs in normal human ciliary body (n=7), cornea (n=7), and trabecualr meshwork (n=7) tissue
Project description:The eye is an intricate organ with limited representation in large-scale functional genomics datasets. The retinal pigment epithelium (RPE) serves vital roles in ocular development and retinal homeostasis. We interrogated the genetics of gene expression of cultured human fetal RPE (fRPE) cells under two metabolic conditions. Genes with disproportionately high fRPE expression are enriched for genes related to inherited ocular diseases. Variants near these fRPE-selective genes explain a larger fraction of risk for both age-related macular degeneration (AMD) and myopia than variants near genes enriched in 53 non-ocular human tissues. Increased mitochondrial oxidation of glutamine by fRPE promoted expression of lipid synthesis genes implicated in AMD. Expression and splice quantitative trait loci (e/sQTLs) analyses revealed shared and metabolic condition-specific loci of each type and several eQTLs not previously described in any tissue. Fine mapping of fRPE e/sQTLs across AMD and myopia genome-wide association data suggests new candidate genes, and mechanisms by which the same common variant of RDH5 contributes to both increased AMD risk and decreased myopia risk. Our study highlights the unique transcriptomic characteristics of fRPE and provides a resource to connect e/sQTLs in a critical ocular cell type to monogenic and complex eye disorders.
Project description:To investigate the function of histone lactylation in ocular melanoma, we analyzed histone lactylation enrichment level in ocular melanoma by CUT&Tag.
Project description:To investigate the function of histone lactylation in ocular melanoma, we analyzed histone lactylation enrichment level in ocular melanoma by CHIP-seq.
Project description:To compare the gene expression profiles of unpassaged, proliferating HUVEC and human iris, retinal and choroidal microvascular endothelial cells. Gene expression profiling revealed significant differences between HUVEC and ocular microvascular endothelial cells suggesting that HUVE cells may not be a suitable surrogate when studying pathophysiological mechanisms of ocular disorders. There were significant differences in the gene expression of important cell signalling pathways in human retinal and choroidal ECs. These differences may be important in the mechanisms and treatment of choroidal and retinal neovascularisation. 12 arrays are included. Endothelial cells were derived from 4 tissues: iris, retina, choroid and human umbilical vein. RNA extracts from cells were hybridised to Affymetrix HGU133plus2 arrays in triplicate.
Project description:To compare the gene expression profiles of unpassaged, proliferating HUVEC and human iris, retinal and choroidal microvascular endothelial cells. Gene expression profiling revealed significant differences between HUVEC and ocular microvascular endothelial cells suggesting that HUVE cells may not be a suitable surrogate when studying pathophysiological mechanisms of ocular disorders. There were significant differences in the gene expression of important cell signalling pathways in human retinal and choroidal ECs. These differences may be important in the mechanisms and treatment of choroidal and retinal neovascularisation.