OC 56.3 - Endothelial Overexpression of Transforming Growth Factor Beta Induced in Chronic Thromboembolic Pulmonary Hypertension

Tuesday, July 12, 2022

11:15 AM – 11:30 AM

Location: ExCel Center, ICC Capital Suite Room 10&11

Presentation Format: In-Person

Presenting Author(s)

MB

Magdalena L. Bochenek, PhD

Post-Doc
Center for Thrombosis and Hemostasis, University Medical Center Mainz, Germany
Mainz, Rheinland-Pfalz, Germany

Co-Author(s)

KS

Kathrin Saar

Post-Doc
Max-Delbrück-Center for Molecular Medicine, Berlin, Germany
Berlin, Berlin, Germany
MN

Maliheh Nazari-Jahantigh

Post-Doc
Institute for Prophylaxis and Epidemiology of Cardiovascular Diseases, Clinic of the University of Munich, Germany
München, Bayern, Germany
CW

Christoph Wiedenroth

Doctor
Kerckhoff Heart and Thorax Center, Department of Thoracic Surgery, Bad Nauheim, Germany
Bad Nauheim, Hessen, Germany
ML

Mareike Lankeit

junior group leader
Center for Thrombosis and Hemostasis, University Medical Center Mainz, Germany
Mainz, Rheinland-Pfalz, Germany
TM

Thomas Münzel

Professor, Head of Cardiology I Department
Center for Cardiology - Cardiology I, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Rhine-Main, Mainz, Germany
Mainz, Rheinland-Pfalz, Germany
EM

Eckard Mayer

Professor
Kerckhoff Heart and Thorax Center, Department of Thoracic Surgery, Bad Nauheim, Germany
Bad Nauheim, Hessen, Germany
SG

Stefan Guth

Head of Department
Kerckhoff Heart and Thorax Center, Department of Thoracic Surgery, Bad Nauheim, Germany
Bad Nauheim, Hessen, Germany
AS

Andreas Schober

Professor
Institute for Prophylaxis and Epidemiology of Cardiovascular Diseases, Clinic of the University of Munich, Germany
München, Bayern, Germany
NH

Norbert Hübner

Professor
Max-Delbrück-Center for Molecular Medicine, Berlin, Germany
Berlin, Berlin, Germany
SK

Stavros Konstantinides

MD, PhD
Center for Thrombosis and Hemostasis, University Medical Center of the Johannes Gutenberg University
Mainz, Rheinland-Pfalz, Germany
Katrin Schäfer, MD

Professor
University Medical Center Mainz, Mainz, Germany
Mainz, Rheinland-Pfalz, Germany

Background: Angiogenesis is a critical event during venous thrombus remodeling, and endothelial cell-poor, fibrotic thrombi are obstructing the pulmonary artery lumen in patients with chronic thromboembolic pulmonary hypertension (CTEPH).

Aims: To examine the genetic footprint of endothelial cells from pulmonary thrombofibrotic material to better understand the non-resolution of thrombus and fibrosis.

Methods: Cells outgrown from pulmonary endarterectomy specimens expressing endothelial cell markers (CTEPH-ECs) were processed for whole gene microarray analysis. Spatial laser microdissection followed by nCounter gene expression analysis and immunohistochemistry of tissue microarrays were used to confirm expression of candidate proteins in CTEPH tissue. Plasma levels of candidate genes (TGFBI, TAGLN, FSTL3 and STC2) were examined in CTEPH patients. Finally, lentiviral overexpression of TGFBI was used to assess a role of TGFBI in the process of angiogenesis.

Results: Out of 26,808 genes examined, 527 were differentially regulated in CTEPH-ECs compared to human pulmonary arterial endothelial cells (HPAECs). Biological pathway analysis and RT2 PCR profiler analysis confirmed that factors downstream of transforming growth factor beta (TGFβ) such as TGFβ-induced (TGFBI) or transgelin (TAGLN), or involved in TGFβ signaling such as follistatin-like 3 (FSTL3), were expressed at significantly higher levels in CTEPH-ECs. Spatial laser microdissection followed by nCounter gene expression analysis and immunohistochemistry of tissue microarrays localized potential disease candidates to vessel-rich regions. Whereas circulating levels of TAGLN and FSTL3 were also increased in patients with pulmonary arterial hypertension, only TGFBI plasma levels were specifically elevated in CTEPH patients and found to decrease after pulmonary endarterectomy. Lentiviral overexpression of TGFBI in HPAECs induced the expression of TAGLN, STC2 and the transcriptional repressor SNAI2, thus phenocopying gene expression patterns in CTEPH-ECs.

Conclusion(s): Our findings strengthen the importance of significant endothelial alterations in the pathophysiology of CTEPH and suggest that overexpression of TGFBI in endothelial cells is causally involved in the thrombofibrosis.