Kidney Week

Abstract: TH-PO1087

ADAMTS12 Promotes Fibrosis by Cleaving HMCN1, Enabling the Activation and Migration of Injury-Responsive Fibroblasts

Session Information

• CKD: Mechanisms - 1
  October 24, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: CKD (Non-Dialysis)

• 2303 CKD (Non-Dialysis): Mechanisms

Authors

• Koch, Lars, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Lars Koch

• Hoeft, Konrad, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Konrad Hoeft, DrMed

• Schumacher, David, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

David Schumacher, MD

• Zhang, Ling, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Ling Zhang, MD

• Tanzer, Maria, The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia

Maria Tanzer

• Schreibing, Felix, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Felix Schreibing, MD

• Long, Qingqing, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Qingqing Long, MD, MS

• Klinkhammer, Barbara Mara, Department of Pathology, RWTH Aachen University, Aachen, Germany

Barbara Mara Klinkhammer, PhD

• Schikarski, Carla Sophie, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Carla Sophie Schikarski

• Maryam, Sidrah, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Sidrah Maryam, MSc

• Krieg, Sarah, Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Aachen, Germany

Sarah Krieg, MSc, PhD

• Peisker, Fabian, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Fabian Peisker

• Schaefer, Gideon J L, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Gideon J L Schaefer

• Menzel, Sylvia, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Sylvia Menzel

• Humphreys, Benjamin D., Institute of Biochemistry and Molecular Biology, RWTH Aachen University, St. Louis, Missouri, United States

Benjamin D. Humphreys, MD, PhD, FASN

• Boor, Peter, Department of Pathology, RWTH Aachen University, Aachen, Germany

Peter Boor, MD, PhD

• Kramann, Rafael, Department of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology and Hypertension), RWTH Aachen University, Aachen, Germany

Rafael Kramann, MD, PhD, FASN

Group or Team Name

• Dept of Medicine 2 (Nephrology, Rheumatology, Clinical Immunology, Hypertension), RWTH Aachen University.

Background

Fibrosis is the uncontrolled replacement of parenchymal tissue with extracellular matrix (ECM) following tissue injury, which impairs organ function. Myofibroblast, which mainly originate from a small perivascular progenitor population marked by Gli1, are considered the major source of ECM deposition. Following injury, they proliferate, migrate to the interstitium, and differentiate into myofibroblasts.

Methods

We conducted genetic fate-tracing to analyze gene expression in Gli1+ cells after unilateral ureteral obstruction (UUO). Adamts12^-/- and wild-type (WT) mice were subjected to UUO and myocardial infarction (MI) surgeries to assess fibrosis, ejection fraction, and spatial gene expression using 10X Visium spatial transcriptomics. In vitro, human renal PDGFRB+ fibroblast cell lines with CRISPR-mediated ADAMTS12 knockout (KO) and lentiviral overexpression of catalytically active and inactive ADAMTS12 were analyzed using bulk RNA sequencing and live-cell imaging. ECM expression of WT and ADAMTS12-KO fibroblasts was characterized by mass spectrometry. Digestion assays and siRNA knockdown assessed the interaction of ADAMTS12 and HMCN1.

Results

Adamts12 was identified as top upregulated gene in Gli1+ fibroblast progenitor cells following UUO. In UUO and MI models, Adamts12^-/- mice exhibited reduced fibrosis and preserved left ventricular ejection fraction post-MI. Spatial transcriptomics post-MI confirmed depletion of injury-response fibroblasts in Adamts12^-/- mice. In vitro, PDGFRb+ fibroblasts expressing catalytically active ADAMTS12 showed a distinct injury-response fibroblast signature and increased migration-associated genes. Live-cell imaging confirmed accelerated cell migration by active ADAMTS12. Mass spectrometry revealed accumulation of HMCN1 in ADAMTS12-KO cell ECM. Digestion assays confirmed HMCN1 cleavage by ADAMTS12, a hitherto unknown ADAMTS12 substrate. Enhanced fibroblast migration with cleaved HMCN1 was validated by siRNA knockdown and live-cell imaging.

Conclusion

We identified ADAMTS12 as a critical checkpoint in fibrosis, that controls fibroblast migration and activation via cleavage of a hitherto unknown substrate, HMCN1.

Funding

• Government Support – Non-U.S.