Single Cell Transcriptomic Analysis of Popliteal Lymphatic Vessels and Peripheral Veins with Smooth Muscle Cell Enrichment in TNF-Tg vs WT Mice

Lymphatic dysfunction exists in tumor necrosis factor transgenic (TNF-Tg) mice and rheumatoid arthritis (RA) patients. While joint-draining TNF-Tg popliteal lymphatic vessels (PLVs) have deficits in contractility during end-stage arthritis, the nature of lymphatic muscle cells (LMCs) and their TNF altered transcriptome remain unknown. Thus, we performed single-cell RNA-sequencing (scRNAseq) on TNF-Tg vs wild-type peripheral vasculature. We utilized strategic enrichment for smooth muscle cells using a Cspg4-Cre;tdTomato reporter model, and through post-hoc analysis we validated the ability to enrich for these tdTomato+ populations based on scatter alone. As such, we evaluated changes in LMC populations vs vascular smooth muscle cells (VSMCs), and dynamic changes in peri-vascular immune cell populations during chronic inflammatory-erosive arthritis. Four total single-cell RNA-seq samples are provided. Two samples are replicates from Cspg4-Cre;tdTomato mice with cell enrichment for tdTomato+ cells (tdT1 and tdT2). Two samples derive from age-matched (9-months-old) WT and TNF-Tg, where the WT serves as control to the TNF-Tg mice with chronic inflammatory-erosive arthritis. The WT and TNF-Tg samples were sorted based on scatter alone. Each sample consists of cell isolates derived from n = 3 mice pooled together, and all mice were on a C57BL6 background.