biostudies-arrayexpressHashem MohammdianHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15133Enthesitis is a characteristic feature of the course of psoriatic arthritis (PsA). To date, most data on enthesitis in PsA have been based on clinical assessment as well as MRI or ultrasound, due to the challenge of obtaining good quality enthesal tissue for molecular analysis. To date, there are no molecular data assessing cell abundance in homeostasis and inflammation at the enthesis site in humans. Therefore, we collected fresh enthesial tissue from cadavers during autopsy and inflamed enthesial tissue from a PsA patient during surgery. Single cell suspensions were obtained by enzymatic dissociation and 3' single cell RNA sequencing was performed on the 10x Genomics platform. Thus, single cell RNA sequencing of these highly specialised tissues may further our understanding of the pathogenesis of PsA.biostudies-arrayexpressNucleic Acid Extraction - 15,000 and 1,000 cells from healthy or inflamed enthesis were loaded into a single well of a Chromium chip G, respectively (10x Genomics) immediately after tissue dissociation and further processed according to standard Chromium Next GEM Single Cell 3' Gene Expression protocol (10x Genomics protocol CG000316 Rev C).Sequencing - Libraries were sequenced as paired end (PE) 150 bp by Illumina sequencing to 65-80% saturation.Sample Collection - Biopsies were taken either at autopsy or during surgery. The instrument was inserted to bone contact and then gently withdrawn to obtain the biopsy (5 mm) 5 mm above the bone. Biopsies were immediately transferred into saline. Entheseal tissues were then minced and digested in RPMI 1640 containing 2 mg ∙ ml−1 collagenase D (Roche), 0.2 mg ∙ ml−1 Dispase II (Sigma-Aldrich) and 0.2 mg ∙ ml−1 DNase I (Roche). Samples were incubated three times at 37 °C for 20 min with constant shaking (2,000 rpm) on a thermal shaker (Eppendorf). After 20 min, the supernatant was collected, filtered (70 µm) and fresh digestion medium was added to the samples. The digestion was repeated for a total of 3 times. RBC lysis was performed using RBC lysis buffer (BioLegend) according to the manufacturer’s recommendations. Cells were washed, counted and concentrated to 1,000 cells / µl.Library Construction - 3’ gene expression library was generated using Chromium Next GEM Single Cell 3' Kit 3.1 Library Construction Kit .OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - . Entheseal scRNAseq datasets were imported into R and converted to Seurat objects. Cells with UMI < 1700, number of genes < 900, mitochondrial gene ratio > 0.2 and complexity < 0.8 were removed. Gene expressed in less than 5 cells were discarded. Seurat Log Normalisation and scaling was performed.Sequence Alignment - Reads were mapped to the GRCh38 human genome (GENCODE v32/Ensembl98) using the 10x Genomics Cell Ranger pipeline (7.2.0) with default settings.UnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNA from single cellsHomo sapiensVladyslav FedorchenkoMario Raphael AngeliAndreas RammingMaria Gabriella RaimondoHashem MohammdianfalseSingle cell RNA-seq of human enthesis from healthy and inflamed tissuesEnthesitis is a characteristic feature of the course of psoriatic arthritis (PsA). To date, most data on enthesitis in PsA have been based on clinical assessment as well as MRI or ultrasound, due to the challenge of obtaining good quality enthesal tissue for molecular analysis. To date, there are no molecular data assessing cell abundance in homeostasis and inflammation at the enthesis site in humans. Therefore, we collected fresh enthesial tissue from cadavers during autopsy and inflamed enthesial tissue from a PsA patient during surgery. Single cell suspensions were obtained by enzymatic dissociation and 3' single cell RNA sequencing was performed on the 10x Genomics platform. Thus, single cell RNA sequencing of these highly specialised tissues may further our understanding of the pathogenesis of PsA.2026-01-01T00:00:00Z2026-01-02T02:02:32.311Z2025-05-20T08:06:10.385ZE-MTAB-15133ERP172743EFO_0002944EFO_0004170EFO_0005684EFO_0004917EFO_0005518EFO_0003816EFO_0004184