Kidney Week

Abstract: TH-PO558

Using Laser Capture Microdissection and Proteomics to Identify Key Components of Metabolic Signaling Pathways in Glomerular Diseases

Session Information

• Glomerular Diseases: Omics, Biomarkers, and Tools
  October 24, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

• 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

• Kroll, Marie-Kristin, University of Cologne Center for Molecular Medicine Cologne, Cologne, Nordrhein-Westfalen, Germany

Marie-Kristin Kroll, MSc

• Volker, Linus A., Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Linus A. Volker, MD

• Höhne, Martin, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Martin Höhne, PhD

• Schermer, Bernhard, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Bernhard Schermer, MD

• Kann, Martin, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Martin Kann, MD

• Benzing, Thomas, Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Thomas Benzing, MD, PhD, FASN

• Brinkkoetter, Paul T., Exzellenzcluster CECAD in der Universitat zu Koln, Koln, Nordrhein-Westfalen, Germany

Paul T. Brinkkoetter, MD

Background

Renal biopsies are classified based on descriptive parameters and validated antibodies. This histopathological assessment does not routinely consider molecular parameters. Here, we aim to unravel the key molecular changes in metabolic signaling pathways to establish a set of molecular markers linked to distinct disease states or response to therapy in glomerular diseases, including MCD and FSGS.

Methods

A technical workflow was established for rodent kidney samples. Glomerular sections of genetically modified mouse models and experimentally treated animals, including the induction of nephrotoxic nephritis (NTN) by serum application, as well as a rodent FSGS model, were isolated by Laser capture microdissection (LCM). Isolated glomerular sections were processed and analyzed using mass spectrometry.

Results

We identified proteins with dissenting regulation within a large-scale proteomics approach using genetically modified and experimentally treated mouse models. The expression patterns of significantly regulated proteins revealed reliable clustering of genetically or experimentally affected mice and control samples. Furthermore, some of the identified proteins correlated with transcriptome data linked to podocyte damage in rodent models.

Conclusion

Our workflow identified significantly regulated proteins and distinct overlaps with transcriptome data in the glomerular sections of mice with renal impairment. Next, this workflow will be applied to human biopsies of FSGS and MCD patients to identify regulated proteins of interest in a large-scale setup. Following, in a targeted proteomics approach key proteins of metabolic signaling pathways will be analyzed in depth. By combining molecular findings with comprehensive clinical background information, including disease state, progression, and responsiveness to therapy, we aim to define a set of marker proteins allowing a precise molecular characterization of patient biopsies and thereby supporting indications for suitable therapies and drug treatment strategies for MCD and FSGS patients.

Funding

• Government Support – Non-U.S.