PB0771 - Platelets promote neutrophil extracellular traps mediated thrombo-inflammation and glomerular endothelial dysfunction in diabetic kidney disease

Monday, July 11, 2022

6:30 PM – 7:30 PM

Location: Exhibition

Presentation Format: In-Person

Presenting Author(s)

AG

Anubhuti Gupta

PhD student
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany

Co-Author(s)

KS

Kunal Singh

PhD student
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany
RY

Ruaa Younis

PhD student
Institute for Laboratory Medicine, Leipzig University
Leipzig, Sachsen, Germany
SK

Shruthi Krishnan

Post Doctoral researcher
University of Leipzig
Leipzig, Sachsen, Germany
Sameen Fatima, MPhil

PhD student
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany
SA

Saira Ambreen

PhD student
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany
HM

Han Mai

PhD researcher
University of Leipzig
Leipzig, Sachsen, Germany
NH

Nikola Hoffmann

Medical student
University of Leipzig
Leipzig, Sachsen, Germany
RB

Ronald Biemann

Project leader
University of Leipzig
Leipzig, Sachsen, Germany
KS

Khurrum Shahzad

Project leader
University of Leipzig
Leipzig, Sachsen, Germany
SZ

Silke Zimmermann

PhD researcher
University of Leipzig
Leipzig, Sachsen, Germany
Berend Isermann, PhD MD

Institute director
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany
SK

Shrey Kohli, PhD

Group leader
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University hospital Leipzig, Leipzig, Germany
Leipzig, Sachsen, Germany

Background: Diabetic kidney disease (DKD) is a major cause of end-stage renal failure and is associated with endothelial dysfunction, platelet-hyperactivity, immune-cell infiltration and dysfunctional glomerular filtration barrier. Mechanistic insights into the role of platelets for DKD progression are limited.

Aims: We aim to scrutinize the mechanistic interplay between platelets and neutrophil extracellular traps (NETs) and ensuing renal thrombo-inflammation in DKD.

Methods: Renal function (albuminuria, fractional mesangial area), platelet activation and NET formation was evaluated in a mouse diabetes model. Therapeutic interventions (Aspirin, Anakinra, Solulin, GSK484) were performed in sub-groups of mice with drugs starting after 16 weeks of diabetes until 24 weeks to study disease reversal. In vitro studies were performed using glomerular endothelial cells (GENCs), platelets and neutrophils exposed to hyperglycaemia under static and flow conditions.

Results: Activated platelets (CD62P) and neutrophil extracellular traps (NETs; CitH3, NE, PAD4) were readily detectable in glomeruli of diabetic mice. Expression of inflammasome markers (NLRP3, IL1β) were positively correlated, reduced thrombomodulin (TM) expression was negatively correlating with NETs. In vitro, platelets exacerbate high glucose and NET induced endothelial dysfunction (p-eNOS, KLF2, KLF4 and TM), sterile inflammation (IL1β, NLRP3), cell death (TUNEL, Caspase-3) and glomerular filtration barrier disruption (enhanced FITC-albumin leakage, disoriented VE cadherin). Under flow conditions, platelets promoted hyperglycaemia induced NET formation on GENCs. Inhibition of platelet activation (Aspirin), amelioration of NETs by inhibition of histone citrullination PAD4 inhibition (GSK484), IL-1 receptor inhibition (anakinra) or restoring TM expression (solulin) ameliorated these effects in vitro and resulted in diabetic kidney disease reversal in vivo.

Conclusion(s): Hyperglycaemia promotes platelet-neutrophil interactions resulting in NETs, hypercoagulability, endothelial sterile inflammation, glomerular endothelial dysfunction, barrier disruption and cell death. These changes aggravate the course of DKD. Inhibition of platelets or NETs is a promising therapeutic strategy for DKD.