VU Research Portal Towards better understanding of the pathophysiology of aortic aneurysms: the role of smooth muscle cells Bogunovic, N. 2020 document version Publisher's PDF, also known as Version of record Link to publication in VU Research Portal citation for published version (APA) Bogunovic, N. (2020). Towards better understanding of the pathophysiology of aortic aneurysms: the role of smooth muscle cells. 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Oct. 2021 TOWARDS BETTER UNDERSTANDING OF THE PATHOPHYSIOLOGY OF AORTIC ANEURYSM AORTIC OF PATHOPHYSIOLOGY THE OF BETTER UNDERSTANDING TOWARDS Towards better understanding Natalija Bogunović was born on February 23rd, 1991 in Belgrade, Serbia. of the pathophysiology of She received her Bachelor’s degree in Biology from aortic aneurysms: the University of Belgrade in 2013. Since she developed an interest in translational research, she the role of smooth muscle cells continued her studies in the Master’s program Experimental Physiology in 2013 at the University of Belgrade. Her interest in cardiovascular diseases spiked during her Master thesis research which she performed at the Institute for Medical Research in Belgrade, where she worked on malignant hypertension. Afterward, she decided to move abroad and continue to pursue her passion for research in cardiovascular diseases by starting her PhD at the VUmc in Amsterdam, as a collaboration between the departments of Vascular Surgery, Physiology, and Clinical Genetics. Natalija’s PhD research was focused on the role of smooth muscle cells in the pathophysiology of aortic aneurysms and the results of that research are presented in this thesis. She is currently working as a postdoc at the Amsterdam UMC, location VUmc and continuing her research into the pathophysiology of aortic aneurysms with a focus on transcriptional regulation. | Natalija Bogunovi Natalija Ć Natalija BogunoviĆ 145647 Bogunovic R13 OMS.indd 2-3 20-08-20 11:41 Towards better understanding of the pathophysiology of aortic aneurysms: the role of smooth muscle cells. Natalija Bogunović 145647 Bogunovic BNW.indd 1 26-08-20 10:57 Towards better understanding of the pathophysiology of aortic aneurysms: the role of smooth muscle cells. ISBN: 978-94-6416-069-7 Financial support by the Dutch Heart Foundation for the publication of this thesis is gratefully acknowledged. The printing of this thesis was kindly supported by Applied Biophysics. The online app was financially supported by Amsterdam Cardiovascular Sciences. The research described in this thesis was supported by the ICaR-AIO grant of the Amsterdam Cardiovascular Science Institute [grant number ICAR-VU AIO 2015]. Copyright © Natalija Bogunović 2020. All rights reserved. No part of this thesis may be reproduced, stored or transmitted in any way or by any means without the prior permission of the author, or when applicable, of the publishers of the scientific manuscripts. Cover image: N. Bogunović Cover layout, layout and printing: Ridderprint | www.ridderprint.nl. 145647 Bogunovic BNW.indd 2 26-08-20 10:57 VRIJE UNIVERSITEIT TOWARDS BETTER UNDERSTANDING OF THE PATHOPHYSIOLOGY OF AORTIC ANEURYSMS: THE ROLE OF SMOOTH MUSCLE CELLS. ACADEMISCH PROEFSCHRIFT ter verkrijging van de graad Doctor of Philosophy aan de Vrije Universiteit Amsterdam, op gezag van de rector magnificus prof.dr. V. Subramaniam, in het openbaar te verdedigen ten overstaan van de promotiecommissie van de Faculteit der Geneeskunde op woensdag 21 oktober 2020 om 11.45 uur in de aula van de universiteit, De Boelelaan 1105 door Natalija Bogunović geboren te Beograd, Servië 145647 Bogunovic BNW.indd 3 26-08-20 10:57 promotoren: prof.dr. J.D. Blankensteijn prof.dr. P.L. Hordijk copromotoren: dr. K.K. Yeung dr. D. Micha 145647 Bogunovic BNW.indd 4 26-08-20 10:57 TABLE OF CONTENTS CHAPTER 1 _____________________________________________________________ 7 Introduction and scope of the thesis ______________________________________ 7 CHAPTER 2 ____________________________________________________________ 25 Transdifferentiation of human dermal fibroblasts to smooth muscle-like cells to study the effect of MYH11 and ACTA2 mutations in aortic aneurysms. _________ 25 Human mutation; 2017; 38(4), 439-450. ___________________________________ 25 CHAPTER 3 ____________________________________________________________ 53 Molecular phenotyping and quantitative assessment of effects of pathogenic variants in aneurysm genes ACTA2, MYH11, SMAD3 and FBN1. _______________ 53 Manuscript in Preparation______________________________________________ 53 CHAPTER 4 ____________________________________________________________ 89 Betaglycan (TGFBR3) upregulation correlates with increased TGF-ß signaling in Marfan patient fibroblasts in vitro. _______________________________________ 89 Cardiovascular Pathology; 2018; 32, 44-49.________________________________ 89 CHAPTER 5 ___________________________________________________________ 109 Impaired smooth muscle cell contractility as a novel concept of abdominal aortic aneurysm pathophysiology. ___________________________________________ 109 Scientific reports; 2019; 9(1), 6837. _____________________________________ 109 CHAPTER 6 ___________________________________________________________ 145 An in vitro method to keep human aortic tissue sections functionally and structurally intact. ____________________________________________________ 145 Scientific reports; 2018; 8(1), 8094. _____________________________________ 145 CHAPTER 7 ___________________________________________________________ 173 Bioengineered Patient-Specific 3D Vascular Scaffolds for the Investigation of Smooth Muscle Cell and Extracellular Matrix Dysfunction in Aortic Aneurysms. 173 Manuscript Submitted ________________________________________________ 173 CHAPTER 8 ___________________________________________________________ 203 Summary and Discussion _____________________________________________ 203 APPENDICES __________________________________________________________ 221 145647 Bogunovic BNW.indd 5 26-08-20 10:57 145647 Bogunovic BNW.indd 6 126-08-20 10:57 CHAPTER 1 CHAPTER 1 Introduction and scope of the thesis Introduction and scope of the thesis 1145647 Bogunovic BNW.indd 7 26-08-20 10:57 Chapter 1 Aortic aneurysms Aortic aneurysms (AA) are pathological dilations of the aorta. The diameter of a healthy aorta is approximately 2 cm, and a dilation can be considered aneurysmal once it exceeds 3 cm, or 150% of the original diameter1. Aneurysms occur in both anatomical segments of the aorta; they are thus accordingly named abdominal aortic aneurysms (AAA) and thoracic aortic aneurysms (ThAA). Figure 1 depicts a schematic comparison between the infrarenal abdominal region of a healthy aorta (Figure 1a) and an infrarenal AAA (Figure 1b). The natural course of the disease is to grow and rupture. Ruptured AA are surgical emergencies associated with an overall mortality rate of up to 90%2, mainly due to internal bleeding complications. Our knowledge about risk factors leading to and/or indicating rupture are restricted, but the most prominent risk factors are large aneurysm diameter and high growth rates of the aneurysmal sac1. AA raises serious concerns in the Figure 1. Schematic representation of an modern age. AAA affects up to 8% of abdominal aortic aneurysm before and men above the age of 651 and 1.4% of after repair. a) Healthy aorta. b) Infrarenal abdominal aortic aneurysm. Elements were 3 women above the age of 75 . ThAA are modified from Servier Medical Art, licensed less common in the general population, under a Creative Common Attribution 3.0 with an incidence of 5-10 individuals Generic License. https://smart.servier.com/; https://creativecommons.org/licenses/by/3.0/ per 100.0004. Many risk factors are associated with AA, but a direct causal relationship is still lacking; the most commonly proposed ones include: gender, smoking, old age, cardiovascular comorbidities and genetic causes. Men are reported to have six times higher risks of developing an AAA5 than women. That risk further increases 40% every five years above the age of 655. Smoking has a strong clinical association with developing an AA6,7. A study reports that 87% of AAA patients in a cohort of Swedish men were either current or former smokers6. Patients suffering from hypertension are reported to have 30-40% more chance of developing an AA. Furthermore, people who were ever treated with antihypertension medication have 80% more risk of developing an AA5. It is, however, noted that the prevalence of AA in the Western world is decreasing with increasing emphasis on healthier lifestyle and better medication for cardiovascular comorbidities, emphasizing the role these comorbidities play in AA pathogenesis8.