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Abstract: SA-OR023

Single-Cell Transcriptomics Analysis of Preaccess and Arteriovenous Fistula Veins

Session Information

Category: Dialysis

  • 803 Dialysis: Vascular Access

Authors

  • He, Yong, University of Florida, Gainesville, Florida, United States
  • Hu, Qiongyao, University of Florida, Gainesville, Florida, United States
  • Cai, Guoshuai, University of Florida, Gainesville, Florida, United States
  • Berceli, Scott A., University of Florida, Gainesville, Florida, United States
Background

An arteriovenous fistula (AVF) vein needs extensive remodeling for use. The cellular and molecular mechanisms of AVF remodeling in ESRD patients remain poorly understood. To address this, we analyzed vein cell-type specific transcriptomics using scRNA-seq.

Methods

Vein samples were collected during the initial creation of brachial artery-basilic vein AVF and the 2nd stage vein transposition surgery 6 weeks later. Cells were extracted by enzymatic digestion. Sequencing library construction used 10X Genomics Chromium Next GEM Single Cell Kit v3.1. Counts output from Cell Ranger were analyzed in Seurat v5.3.0.

Results

38 (21 preaccess) vein samples were collected from 22 patients. 17 patients had paired samples. 92760 cells were kept after quality check and filtering. The most abundant cells were endothelial cells (EC), myofibroblast (MFB), fibroblast (FB), smooth muscle cells (SMC), pericytes, macrophages/monocytes, and T/NK cells. Except for pericytes, there were no statistically significant differences in proportions of these major cell types between veins and AVFs. There were more pericytes in AVFs. One EC subcluster, accounting for 10% of ECs in veins or AVFs, enriched in genes regulating key processes of AVF remodeling, such as response to fluid shear stress, inflammatory response, coagulation, endothelial-mesenchymal transition, cell-matrix adhesion, wound healing, angiogenesis, and collagen and proteoglycan synthesis in AVFs compared to veins. Contractile SMCs in AVFs had lower gene expressions of contractile apparatus than in veins. One SMC subcluster, accounting for 13% of SMCs in veins or AVFs, enriched in glycosaminoglycan synthesis enzyme genes but had diminished cellular response to hypoxia and detoxification of reactive oxygen species in veins and AVFs than other subclusters. These cells in AVFs also had lower cell-matrix adhesion and cell mobility than in veins. MFBs in AVFs had higher collagen expressions than in veins. The largest FB subcluster expressed higher collagen genes in AVFs than in veins. Proinflammatory macrophages in AVFs had stronger cell surface interactions at the vascular wall than in veins.

Conclusion

With the transcriptome profiling of a large number of vein cells, we identified cell subclusters and their specific transcriptomic changes that were associated with AVF remodeling, offering new insights into the underlying mechanisms.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)