University of Groningen Endothelial plasticity in fibrosis and epigenetics as a therapeutic target Hulshoff, Melanie

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University of Groningen

Endothelial plasticity in fibrosis and epigenetics as a therapeutic target

Hulshoff, Melanie

DOI:

10.33612/diss.146265795

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Publication date: 2020

Link to publication in University of Groningen/UMCG research database

Citation for published version (APA):

Hulshoff, M. (2020). Endothelial plasticity in fibrosis and epigenetics as a therapeutic target. University of Groningen. https://doi.org/10.33612/diss.146265795

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546222-L-bw-Hulshoff 546222-L-bw-Hulshoff 546222-L-bw-Hulshoff 546222-L-bw-Hulshoff Processed on: 27-10-2020 Processed on: 27-10-2020 Processed on: 27-10-2020

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APPENDICES

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ABBREVIATIONS

α-SMA alpha smooth muscle actin AAC ascending aortic constriction AAV adeno associated virus

AcSDKP n-acetyl-seryl-aspartyl-lysyl-proline ADMA asymmetric dimethylarginine ANG-2 angiopoietin-2

ATII angiotensin II

AVC atrio-ventricular canal BMP bone morphogenetic protein CD31 cluster of differentiation 31 ChIP chromatin immunoprecipitation circRNA circular RNA

CKD chronic kidney disease dCas9 deactivated Cas9

dHFCas9 deactivated high-fidelity Cas9 DNMT dna methyltransferase

EMT epithelial-to-mesenchymal transition EndMT endothelial-to-mesenchymal transition eNOS endothelial nitric oxide synthase EZH2 enhancer of zeste homolog 2 FACS fluorescence-activated cell sorting FGF fibroblast growth factor

GAS5 growth arrest-specific 5 GFP green fluorescent protein

H3 histone 3

HCAEC human coronary artery endothelial cells HDAC histone deacetylase

HIF1α hypoxia inducible factor-1α

HUVECs human umbilical vein endothelial cells IL interleukin

lncRNA long non-coding RNA ITR inverted terminal repeat KLF7 kruppel-like factor 7

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ABBREVIATIONS

407

APP

MALAT1 metastasis-associated lung adenocarcinoma transcript 1

MAPK mitogen-activated protein kinase MCEC mouse cardiac endothelial cells MEndT mesenchymal-to-endothelial transition MeDIP methylated DNA immunoprecipitation

miR microRNA

miRNA microRNA

MMP matrix metalloproteinase

NF-κB nuclear factor kappa-light-chain-enhancer of activated B cells NLRP3 NACHT, LRR, and PYD domain-containing protein 3

NO Nitric oxide

NOX nicotinamide adenine dinucleotide phosphate oxidase OFT outflow tract

RAAS renin-angiotensin-aldosterone system RASAL1 ras-gap-like protein 1

RFP red fluorescent protein ROS reactive oxygen species RXFP relaxin family peptide receptor S100A4 s100 calcium-binding protein A4 SARA smad anchor for receptor activation SBEs SMAD-binding elements

TAC transverse aortic constriction TALE transcription activator-like effectors

TET ten-eleven translocation methylcytosine dioxygenase TET3CD TET3 catalytic domain

TGF-β transforming growth factor beta TNF-α tumor necrosis factor alpha TSP-2 thrombospondin-2 TSS transcription start site T2D type 2 diabetes

UUO unilateral ureteral obstruction VE-Cadherin vascular endothelial cadherin VEGF vascular endothelial growth factor ZFP zinc finger protein

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ACKNOWLEDGEMENTS

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ACKNOWLEDGEMENTS

First of all, I would like to thank my supervisors Elisabeth Zeisberg and Guido Krenning. Elisabeth, thank you for the possibilities you gave to me, thank you for believing in me, and thank you for your ultimate support. Your leadership has been a great inspiration. Thank you. Guido, thank you for promoting my independency, thank you for your trust and thank you for your support. Your great scientific insights have always inspired me. Thank you.

Also, I would like to thank my promotor Marco Harmsen for his trust and for acquainting me with the field of endothelial plasticity. I would like to thank Michael Zeisberg for his great scientific insights as well as for his ultimate support. And, I would like to acknowledge Steven Johnsen for his support and scientific insights during my thesis committee meetings.

I am grateful to the manuscript assessment committee, consisting of Prof. Steven A. Johnsen, Prof. Bernd Wollnik, Prof. Jan-Luuk Hillebrands and Prof. Marie-José T. H. Goumans, for taking the time and effort to assess this thesis. I am also grateful to the examining committee, consisting of Prof. Steven A. Johnsen, Prof. Bernd Wollnik, Prof. Jan-Luuk Hillebrands, Prof. Marie-José T. H. Goumans, Prof. Dr. Viacheslav O. Nikolaev, Prof. André Fischer, Prof. Robert H. Henning and Prof. Marianne G. Rots for taking the time and effort to assess the thesis defence.

Thank you to all lab members in Göttingen for providing a great scientific environment. In particular, I would like to thank Xingbo Xu for his great scientific input, for giving me opportunities and for believing in me. Thank you. I am grateful to Xiaoying Tan for her support during my PhD. Also, I would like to acknowledge Gunsmaa Nyamsuren, Sandip Rath, Sabine Maamari and Tim Wilhelmi for their support in the lab. I am also grateful to Sarah Rinkleff, Annika Erdmann and Anika Krueger for their technical assistance.

I would also like to thank all lab members in Groningen who supported me whenever necessary. In particular, I would like to thank Byambaa Nyamsuren and

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APPENDICES

412

Jolien Fledderus for their support in the lab. The Topmaster program Medical and Pharmaceutical Drug Innovation in Groningen stands out for its scientific education which is focused on becoming a top scientist. This training was essential for the successful completion of this PhD, for which I am very grateful.

Of course, I would also like to acknowledge the financial support of this PhD. I am grateful to the Graduate School of Medical Sciences in Groningen who provided a PhD scholarship. Also, I would like to thank the U4 network, and in particular Marco Lange, for financially supporting the exchange between Groningen and Göttingen. And last but not least, I would like to thank the lab in Göttingen as well as in Groningen for their financial support.

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ACKNOWLEDGEMENTS

415

CURRICULUM VITAE &

PUBLICATION LIST

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APPENDICES

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CURRICULUM VITAE

The author of this thesis, Melanie Hulshoff, was born on December 12th 1992 in Groningen, The Netherlands. She studied a Bachelor program in Biomedical Sciences at the University of Groningen, The Netherlands which she completed with distinction. She was also enrolled in the so-called Honours College, an additional extracurricular program for talented students. She then continued with the selective Topmaster program in Medical and Pharmaceutical Drug Innovation at the University of Groningen, which she completed with distinction as well. After completing her Master studies, she was selected for a joint double degree PhD Scholarship in which she could perform her self-designed research proposal. This PhD work was performed at the University of Groningen and the University of Göttingen, Germany under supervision of Dr. Guido Krenning and Prof. Elisabeth M. Zeisberg respectively. During her PhD she has published and prepared several manuscripts on the topic of fibrosis, epigenetics and endothelial plasticity.

PUBLICATION LIST

Xu X, Tan X, Hulshoff MS, Wilhelmi T, Zeisberg M, Zeisberg EM. Hypoxia-induced endothelial-mesenchymal transition is associated with RASAL1 promoter hypermethylation in human coronary endothelial cells.

FEBS Lett. 2016, 590(8):1222-33.

Xu X, Tan X, Tampe B, Wilhelmi T, Hulshoff MS, Saito S, Moser M, Kalluri R, Hasenfuss G, Zeisberg EM, Zeisberg M. High-fidelity CRISPR/Cas9-based gene-specific hydroxymethylation rescues gene expression and attenuates renal fibrosis.

Nat Commun. 2018, 9(1):3509.

Hulshoff MS, Xu X, Krenning G, Zeisberg EM. Epigenetic regulation of

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CURRICULUM VITAE AND PUBLICATION LIST

417

APP

methylation and non-coding RNAs.

Arterioscler Thromb Vasc Biol. 2018, 38:1986-96.

Hulshoff MS, Rath S, Xu X, Zeisberg M, Zeisberg EM. Causal connections from

chronic kidney disease to cardiac fibrosis. Semin Nephrol. 2018, 38(6):629-636.

Hulshoff MS*, Del Monte-Nieto G*, Kovacic J, Krenning G. Non-coding RNA in

endothelial-to-mesenchymal transition. Cardiovasc Res. 2019, 115(12):1716-1731.

Wilhelmi T, Xu X, Tan X, Hulshoff MS, Maamari S, Sossalla S, Zeisberg M and Zeisberg EM. Serelaxin alleviates cardiac fibrosis through inhibiting endothelial-to-mesenchymal transition via RXFP1.

Theranostics. 2020, 10(9):3905-3924.

Xu X*, Hulshoff MS*, Tan X, Zeisberg M, Zeisberg EM. CRISPR/Cas derivatives as novel gene modulating tools: possibilities and in vivo applications.

Int J Mol Sci. 2020, 21(9). pii:E3038.

Xu X*, Hulshoff MS*, Tan X*, Maamari S, Krenning G, Mueller O, Hasenfuss G, Zeisberg M, Zeisberg EM. Split-intein mediated AAV delivery of dCas9-TET3 as antifibrotic therapy.

Submitted.

Hulshoff MS*, Schellinger IN*, Xu X, Fledderus J, Rath SK, Wong FC, Krenning G,

Raaz U, Zeisberg EM. miR-132-3p and KLF7 as novel potential regulators of aortic stiffening-associated EndMT in type 2 diabetes.

Submitted.

Hulshoff MS*, Xu X*, Kühnisch J, Tan X, Zeisberg M, Krenning G, Klaassen S,

Zeisberg EM. Stage-specific mapping of the endothelial-to-mesenchymal transition. In preparation.