biostudies-arrayexpressCheng-Chieh HsuMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16375Clonal haematopoiesis of indeterminate potential (CHIP) involves age-related acquisition and expansion of genes frequently mutated in haematologic malignancies (e.g. DNMT3A, TET2, or JAK2).  JAK2 heightens cardiovascular disease (CVD) risk. The mechanism, although incompletely understood, involves pyroptosis and plasma membrane rupture, mediated by Ninjurin-1 (NINJ1), followed by release of damage-associated molecular patterns and cytokines. Moreover, individuals with JAK2-CHIP have elevated levels of circulating interleukin-17 receptor-A (IL-17RA). IL-17RA signalling, implicated in autoimmunity, paradoxically may have a protective role in atherogenesis. IL-17A produced by T helper 17 (Th17) cells binding to IL-17RA in myeloid cells may induce a TREM2 macrophage response to create a feedback loop and dampen further inflammation. The proteomic association between JAK2-CHIP and IL-17RA, along with the potential role of IL-17RA signalling in atherosclerosis, led us to hypothesize that IL-17RA signalling modifies CVD risk among individuals with JAK2-CHIP.biostudies-arrayexpressNucleic Acid Extraction - RNA was extracted using the 10x Genomics Chromium Single Cell 3'v4 protocol.Sequencing - The prepared library was sequenced using NovaSeq 6000. Sequenced data were demultiplexed to generate the FASTQ files.Growth Protocol - Ldlr-/- recipients were irradiated, then transplanted with bone marrow from mice with Mx1-Cre driving Jak2V617F +/- Ninj1-/-, 4 weeks later ldlr-/- mice were subjected to a western-type diet (Teklad TD88137) for 12 weeksSample Treatment - After 4 weeks of recovery at bone marrow transplantation, mice were subjected to a 12-week Western-type diet (Teklad TD88137)Library Construction - Libraries were prepared using the 10x Genomics Chromium Single Cell 3’ Reagent Kits v4 according to the manufacturer’s instructions.Sample Collection - Aortas were mechanically cleaned and then separated into intimal-medial and adventitial layers. Samples were diced and digested with Liberase, hyaluronidase, and Dnase in media for 45 minutes at 37C. Cells were then washed, blocked with FC block, and hashtaged with TotalSeq-B antibodies and stained for Sytox blue and CD45. Samples were then sorted using flow cytometry (live, CD45+) and subjected to the scRNA-Seq pipelineOrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - FASTQ files were processed and demultiplexed by 10X Cell Ranger v.8.0.1. Reads were aligned to the mouse reference genome build GRCm39 and transcriptome based on Ensembl 110 annotation.Unique molecular identifier (UMI) count matrices were generated by Cell Ranger.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNA from single cellsMus musculusInterleukin-17 receptor-A signalling: atheroprotective role in JAK2 clonal haematopoiesisCheng-Chieh HsuKun Zhao, Cheng-Chieh Hsu, Yash Pershad, Caitlyn Vlasschaert, Robert W Corty, Nan Wang, Jonathan Brett Heimlich, Alan R Tall, Alexander G BickAlan TallfalseThe role of Ninjurin-1 deficiency in Jak2V617F clonal hematopoiesis mediated atherosclerosis plaque progressionClonal haematopoiesis of indeterminate potential (CHIP) involves age-related acquisition and expansion of genes frequently mutated in haematologic malignancies (e.g. DNMT3A, TET2, or JAK2).  JAK2 heightens cardiovascular disease (CVD) risk. The mechanism, although incompletely understood, involves pyroptosis and plasma membrane rupture, mediated by Ninjurin-1 (NINJ1), followed by release of damage-associated molecular patterns and cytokines. Moreover, individuals with JAK2-CHIP have elevated levels of circulating interleukin-17 receptor-A (IL-17RA). IL-17RA signalling, implicated in autoimmunity, paradoxically may have a protective role in atherogenesis. IL-17A produced by T helper 17 (Th17) cells binding to IL-17RA in myeloid cells may induce a TREM2 macrophage response to create a feedback loop and dampen further inflammation. The proteomic association between JAK2-CHIP and IL-17RA, along with the potential role of IL-17RA signalling in atherosclerosis, led us to hypothesize that IL-17RA signalling modifies CVD risk among individuals with JAK2-CHIP.2025-12-22T00:00:00Z2026-05-27T15:26:25.244Z2025-12-11T11:50:35.607ZE-MTAB-1637540986427ERP186422EFO_0002944EFO_0004170EFO_0003789EFO_0005684EFO_0005518EFO_0003816EFO_0004184EFO_000396910.1093/eurheartj/ehaf737