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Single Cell Analysis Reveals Endothelial Activation of Interleukin-6 Mediated Signaling Pathway in Microvascular Inflammation with and without c4d

A. S. Naik1, R. Menon1, E. Farkash1, E. Otto1, C. C. Berthier1, S. Norman1, M. Samaniego2, M. Kretzler1

1University of Michigan, Ann Arbor, MI, 2Henry Ford Hospital, Detroit, MI

Meeting: 2020 American Transplant Congress

Abstract number: A-355

Keywords: Endothelial cells, Kidney transplantation, Rejection

Session Information

Session Name: Poster Session A: Acute Rejection

Session Type: Poster Session

Date: Saturday, May 30, 2020

Session Time: 3:15pm-4:00pm

 Presentation Time: 3:30pm-4:00pm

Location: Virtual

*Purpose: Microvascular inflammation (MVI) is a feature of antibody mediated rejection. MVI can be seen with or without activation of other effectors of antibody injury such as the complement system. How Endothelial Cell (EC) injury patterns differ across EC beds (glomerular,peritubular) and in respect to complement activation remains unclear. We hypothesized that ScRNA-seq will allow us to study EC activation states to identify drivers of injury thereby allowing for precise therapeutic targeting.

*Methods: We leveraged our surveillance and indication biopsy program to identify 2 patients with MVI (g+ptc >1) without c4d deposition (Group 1) or with c4d deposition (Group 2). 3 living donor pre-perfusion biopsies served as normal controls. ScRNA-seq was performed on a 10X chromium platform and processed per KPMP protocols. Cell-type-specific markers were used for cell clustering. Enrichment analysis for biological pathways was performed with Enrichr tool using NCI 2016 pathway interaction database.

*Results:

Clinical Characteristics
g score ptc score c4d antiHLA DSA Surveillance vs. Indication biopsy
Group1: Pt1 2 2 0 No Surveillance
Group 1: Pt 2 1 2 0 Yes Surveillance
Group 2: Pt 1 1 1 1 Yes Surveillance
Group 2: Pt2 1 2 1 Yes Indication

Differentially expressed genes (DEG) between Glomerular Endothelial Cells (GEC) and Peritubular endothelial Cells (PTECs) in controls vs. both MVI groups revealed enrichment of genes associated with IL6 signaling pathway.

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DEG between MVI groups revealed enrichment of genes associated with VEGF/VEGFR and CD40/CD40L signaling in GECs. Activation of genes associated with MAPK, CXCR2 and TNF receptor signaling was found among PTECs.

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*Conclusions: scRNA-seq analysis reveals evidence of endothelial IL6 pathway activation in biopsies with MVI independent of c4d status, suggesting a common pathway being activated across both phenotypes. There were however differences in EC activation states by capillary beds across both groups.

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To cite this abstract in AMA style:

Naik AS, Menon R, Farkash E, Otto E, Berthier CC, Norman S, Samaniego M, Kretzler M. Single Cell Analysis Reveals Endothelial Activation of Interleukin-6 Mediated Signaling Pathway in Microvascular Inflammation with and without c4d [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/single-cell-analysis-reveals-endothelial-activation-of-interleukin-6-mediated-signaling-pathway-in-microvascular-inflammation-with-and-without-c4d/. Accessed May 10, 2025.

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