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Identification and characterization of novel peroxisomal disorders

Falkenberg, K.D.

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Citation for published version (APA):Falkenberg, K. D. (2018). Identification and characterization of novel peroxisomal disorders.

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Download date: 04 Jan 2020

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Chapter 7

Appendix

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Authors’ Contributions Chapter 2: ACBD5 Deficiency Causes a Defect in Peroxisomal Very

Long-Chain Fatty Acid Metabolism

KDF, JK, PM, RJ and CWTR performed laboratory analyses. AP and AV were

responsible for the clinical care of the patient, provided the patient’s clinical

information and revised the manuscript. SF supervised the study and wrote the

manuscript. HRW supervised the study and revised the manuscript. KDF, MS and

RJAW revised the manuscript.

Chapter 3: Allelic Expression Imbalance Promoting a Mutant PEX6

Allele Causes Zellweger Spectrum Disorder

KDF performed genetic and functional studies. KDF and HRW designed genetic

and functional studies. HRW, NEB, SJS, MR, AP and ABM performed Sanger-

sequencing and/or Next Generation Sequencing and independently identified the

PEX6-p.Arg860Trp variant. NEB, ABM, FCCK, AS, ME, KN, FvS, MERG, IKK and

SF contributed clinical and biochemical data and/or fibroblasts of patients. RJAW

and HRW supervised the project. KDF and HRW wrote the manuscript. NEB, ABM,

AS and SF revised the manuscript.

Chapter 4: Heimler Syndrome Is Caused by Hypomorphic Mutations

in the Peroxisome-Biogenesis Genes PEX1 and PEX6

IR, NA-S, SG, GV, JEU, KEC, SGW, NAR, MEA, GCB, HR, AH, AF, SL, NC, KRO,

CELS, CFI, AJM, CE, JAP, MT, SJD, AS, and AAM provided clinical data and patient

material and/or performed Whole Exome Sequencing analyses. KDF performed

functional assays. SF supervised laboratory diagnostic analyses. KDF, MS, WGN,

HRW and GVC wrote and revised the manuscript. WGN, HRW and GVC supervised

the project.

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Chapter 5: Autophagy Inhibitors Do Not Restore Peroxisomal

Functions in Peroxisome Biogenesis Disorders

FCCK and KDF designed and performed experiments, and wrote the manuscript.

RO designed experiments and revised the manuscript. JK performed experiments.

SF supervised laboratory analyses. RJA supervised the study. HRW supervised the

study, designed experiments and revised the manuscript.

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Authors’ AffiliAtions

University of Amsterdam, Amsterdam, the Netherlands

Laboratory Genetic Metabolic Diseases, Academic Medical Center

KD FalkenbergRJA Wanders, PhDHR Waterham, PhDFCC Klouwer, MDR OfmanS Ferdinandusse, PhDJ KosterPA MooyerCWT van Roermund, PhD

Kennedy Krieger Institute, Baltimore, Maryland, USA

R Jones, PhDAB Moser

McGill University, Montreal, Quebec, Canada

Department of Pediatrics and Human Genetics,

NE Braverman, MD

Children’s National Health System, Washington DC, USA

Department of Neurology

A PizzinoA Vanderver, MD

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University of Exeter, Exeter, Devon, UK

College of Life and Environmental Sciences, Biosciences

M Schrader, PhD

Johns Hopkins University School of Medicine, Baltimore, MD, USA

Institute of Genetic Medicine and Department of Neurology,

SJ Steinberg, PhD

Bellvitge Institute for Biomedical Research (IDIBELL), Barcelona, Spain

Neurometabolic Diseases Laboratory, Institute of Neuropathology

A Schlüter, PhDM Ruiz, PhDA Pujol, MD, PhD

Karolinska University Hospital, Stockholm, Sweden

Centre for Inherited Metabolic Diseases

M Engvall, MD, PhDK Naess, MD, PhD

University of Groningen, University Medical Center Groningen, Beatrix Children’s Hospital, Groningen, The Netherlands

Department of Pediatrics

F van Spronsen, MD, PhD

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Maastricht University Medical Center, Maastricht, The Netherlands

Department of Pediatrics

I Körver-Keularts, PhDME Rubio-Gozalbo, MD, PhD

Université Mohammed V, Rabat, Morocco

Centre de Génomique Humaine, Faculté de Médecine et de Pharmacie,

I Ratbi, MDS Guaoua, PhDA Sefiani, MD

Département de Pédodontie-Prévention, Faculté de Médecine Dentaire,

M El Alloussi, MDH Ramdi

University of Antwerp, Antwerp, Belgium

Department of Medical Genetics

M SommenG van Camp, PhDG Vandeweyer, PhD

St. Mary’s Hospital, Manchester Academic Health Sciences Centre, Manchester, UK

Manchester Centre for Genomic Medicine

WG Newman, PhDN Al-SheqaihJE Urquhart, MDKE Chandler, MDSG Williams, MDNA Roberts, PhDGC Black

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University of Manchester, Manchester, UK

Faculty of Life Sciences

AA Mironov, MD, PhD

Schneider Children’s Hospital of Long Island Jewish Medical Center, New Hyde Park, NY, USA

Division of Human Genetics

A Heimler

Liverpool Women’s NHS Foundation Trust, Liverpool, UK

Department of Clinical Genetics

A Fryer, MD

Our Lady’s Children’s Hospital, Crumlin, Dublin, Ireland

National Centre for Medical Genetics

S-A Lynch, MD

Birmingham Women’s Hospital NHS Trust, Birmingham, UK

West Midlands Regional Genetics Service

N Cooper, MDKR Ong, MD

St. James’s University Hospital, University of Leeds, Leeds, UK

Leeds Institute of Biomedical and Clinical Sciences

CEL Smith, PhDCF Inglehearn, AJ Mighell, PhDJA Poulter, PhD

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School of Clinical Dentistry, University of Sheffield, UK

Academic Unit of Oral Health and Development,

C Elcock, PhD

University of Cambridge, Cambridge, UK

Department of Medical Genetics and National Institute for Health Research Cambridge Biomedical Research Centre

M Tischkowitz, PhD

University Hospital of Wales, Cardiff, UK

Institute of Medical Genetics

SJ Davies

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AbbreviAtions

3-MA 3-methyladenine

AAA+ ATPases ATPases Associated with diverse cellular Activities

ABCD1 ATP-binding cassette sub-family D 1 (alternatively known as ALDP)

ABCD2 ATP-binding cassette sub-family D 2 (alternatively known as ALDRP)

ABCD3 ATP-binding cassette sub-family D 3 (alternatively known as PMP70)

ACAA1 3-ketoacyl-CoA thiolase

ACBD5 Acyl-CoA binding domain containing protein 5

ACBP Acyl-CoA binding protein

ACOX1 Acyl-CoA oxidase 1

ACOX2 Branched chain acyl-CoA oxidase

ActD Actinomycin D

AEI Allelic expression imbalance

AGPS Alkyl-glycerone-phosphate synthase (formerly known as alkyl-dihydroxyacetone phosphate synthase or ADHAPS)

AMACR α-methylacyl-CoA racemase

ATP Adenosin-Triphosphate

BAAT Bile acid-CoA:amino acid N- acyltransferase

bp Basepairs

C26:0 Cerotic acid

CA Cholic acid

CDCA Chenodeoxycholic acid

cDNA Copy- or complement-DNA

CNS Central nervous system

CoA Coenzyme A

CPT1 Carnitine palmitoyl-CoA transferase

CQ Chloroquine

CRISPR Clustered Regularly Interspaced Short Palindromic Repeats

DNA Desoxy-ribonucleic acid

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D2 domain ATPase domain (in AAA ATPases)

D3 Deuterated

DBP D-bifunctional protein (alternatively known as multifunctional protein 2, MFP2)

DHAP Dihydroxyacetone phosphate

DHCA Dihydroxycholestanoic acid

DMA Dimethyl acetal

DMEM Dulbecco’s modified Eagle medium

EM Electron microscopy

ER Endoplasmic reticulum

ESI Electrospray ionization

ExAc Exome Aggregation Consortium

FAR1 Fatty acyl-CoA reductase 1

FDA Food and Drug Administration

fw Forward

GATK Genome Analysis Toolkit

gDNA Genomic DNA

GFP Green fluorescent protein

Gly Glycerol

GNPAT Glycerone-phosphate O-acyltransferase (formerly known as dihydroxyacetone phosphate acyltransferase or DHAPAT)

HACL1 2-hydroxyacyl-CoA lyase

HCQ Hydroxychloroquine

HS Heimler syndrome

HUGO Human Genome Organisation

Indel Insertions/deletions

kb Kilobases

kDa Kilo Dalton

L-arg L-arginine

LBP L-bifunctional protein (alternatively known as multifunctional protein 1, MFP1)

LC Liquid chromatography

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lncRNA Long non-coding RNA

MAF Minor allele frequency

miR, miRNA MicroRNA

mRNA Messenger RNA (ribonucleic acid)

MS Mass spectrometry

MRI Magnetic resonance imaging

MRM Multiple reaction monitoring

NAD Nicotinamide adenine dinucleotide

NGS Next Generation Sequencing

PAM Protospacer adjacent motif

PAS Polyadenylation site

PBDs Peroxisome biogenesis disorders

PBD-ZSDs Peroxisome biogenesis disorders within the Zellweger spectrum

PC Phosphatidylcholine,

PEX5L Long isoform of PEX5

PHYH Phytanoyl-CoA hydoxylase

PIPOX L-pipecolate oxidase

PMP Peroxisomal membrane protein

POLII RNA Polymerase II-transcription machinery

PrDH Pristanal dehydrogenase

PTS Peroxisomal targeting signal

PXMP2 Peroxisomal membrane protein 2

RACE Rapid amplification of cDNA ends

RCDP Rhizomelic Chondrodysplasia Punctata

RD Refsum Disease

RT Realtime

rev Reverse

SCPx Sterol carrier protein-X thiolase

SD Standard deviation

SDS Sodium dodecyl sulfate

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SEDs Single Enzyme Deficiencies

SEM Standard error of mean

SKL Peroxisomal targeting signal serine-lysine-leucine (alternatively known as PTS1)

SNHL Sensorineural hearing loss

SNP Single nucleotide polymorphism

SNV Single nucleotide variant

SRH Second region of homology (in ATPases)

ssODN Single-stranded oligo oligodeoxynucleotides

STR Short tandem repeats

TF Transcription factor

THCA Trihydroxycholestanoic acid

UPLC Ultra performance liquid chromatography

UTR Untranslated region

VAPA / VAPB Vesicle-associated membrane protein-associated protein A / B

VLCFAs Very long-chain fatty acids

WES Whole Exome Sequencing

X-ALD X-linked adrenoleukodystrophy

yo Years old

ZSDs Zellweger spectrum disorders

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CurriCulum vitAe

Kim Desiree Falkenberg was born in 1988 in Bocholt, Germany. She finished

secondary school at the St.-Josef Gymnasium in Bocholt, Germany, with an ‘Abitur’

degree in 2007 and started her Bachelor studies in ‘Biosciences’ at the University

of Münster in the same year. After graduating in 2010 she continued her studies

in ‘Biosciences’ at the University of Münster in a M.Sc. course. During this master

course, she spent one semester abroad at the Norwegian University of Science and

Technology (NTNU) in Trondheim, Norway. In 2013, she received her master’s

degree and began her Ph.D. training as an Early Stage Researcher in the Marie

Curie Initial Training Network PerFuMe (Peroxisome Function and Metabolism)

at the Laboratory Genetic Metabolic Diseases at the Academic Medical Center in

Amsterdam, the Netherlands, under supervision of her promotors Hans R. Waterham

and Ronald J.A. Wanders. For four years, she investigated the molecular mechanisms

of peroxisomal diseases, the results of which are presented in this thesis.

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PortfolioYear Work-

load (ECTS)

Courses & workshops

Scientific and laboratory skillsPractical Biostatistics 2013 1.1

Bioinformatics Sequence Analysis 2014 1.1

PerFuMe Workshop I “Systems Biology of metabolic pathways”, Wageningen, NL

2014 1.0

PerFuMe Workshop II “Peroxisomes in Health and Disease”, Exeter, UK

2014 1.0

Bioinformatics 2015 1.1

DNA Technology 2015 2.1

Basic Microscopy 2015 1.6

Advanced qPCR 2016 0.7

Transferable skillsAMC World of Science 2013 0.7

Language Course Dutch (beginners level) 2013

Language Course Dutch (intermediate level) 2014

Oral Presentation in English 2014 0.8

Project Management 2015 0.6

Scientific Writing 2015 1.5

PerFuMe Workshop III “Science in Industry”, Potsdam, DE 2015 1.0

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Presentations

PerFuMe Kick-off conference, Groningen, NLPoster presentation & elevator pitch

2013 0.5

Annual ACM/MDL Meeting, Lunteren, NLPoster presentation

2014 0.5

Marie Sklodowska-Curie Conference (ESOF), Copenhagen, DKPoster presentation

2014 0.5

4th Open European Peroxisome Meeting (OEPM), Neuss, DEOral Presentation

2014 0.5

Annual ACM/MDL Meeting, Lunteren, NLOral Presentation

2015 0.5

Annual ACM/MDL Meeting, Lunteren, NLOral Presentation

2015 0.5

2nd International conference “Peroxisome Formation, Function and Metabolism”, Hamburg, DE

Oral presentation

2016 0.5

5th Open European Peroxisome Meeting (OEPM), Vienna, ATOral presentation

2016 0.5

Attended conferences

PerFuMe Kick-off conference, Groningen, NL 2013 1.0

Marie Skłodowska-Curie Conference: ESOF, Copenhagen, DK 2014 0.5

ACM/MDL Meeting 2014, Lunteren, NL 2014 1.0

4th Open European Peroxisome Meeting (OEPM), Neuss, DE 2014 0.75

ACM/MDL Meeting 2015, Lunteren, NL 2015 1.0

AG&M Meeting 2016, Garderen, NL 2016 1.0

2nd International conference “Peroxisome Formation, Function and Metabolism”, Hamburg, DE

2016 1.0

5th Open European Peroxisome Meeting (OEPM), Vienna, AT 2016 0.75

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Other PhD training

Progress meetings PerFuMe 2013-2016

1.0

Research meetings at Laboratory Genetic Metabolic Diseases, Academic Medical Center Amsterdam, NL

Oral presentations

2013-2017

2.0

Journal club at Laboratory Genetic Metabolic Diseases, Academic Medical Center Amsterdam, NL

Oral presentations

2013-2017

2.0

Secondment I, Exeter, UK 2015 2.5

Secondment II, Sevilla, ES 2015 2.5

Parameters of Esteem

Best Abstract Award, 2nd prizeAG&M Meeting, Garderen, NL

2016

Best Speaker Award, 1st prize2nd International conference “Peroxisome Formation, Function

and Metabolism”, Hamburg, DE

2016

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PubliCAtions * equal contribution

Ratbi I*, Falkenberg KD*, Sommen M* et al.

“Heimler Syndrome Is Caused by Hypomorphic Mutations in the Peroxisome-Biogenesis Genes PEX1 and PEX6.”

Am. J. Hum. Genet. 97, 535-545 (2015). doi:10.1016/j.ajhg.2015.08.011.

Ferdinandusse S*, Falkenberg KD* et al.

“ACBD5 deficiency causes a defect in peroxisomal very long-chain fatty acid metabolism.”

J. Med. Genet. 54(5):330-337 (2017). doi:10.1136/jmedgenet-2016-104132.

Bjørgo K, Fjær R, Mørk HH, Ferdinandusse S, Falkenberg KD et al.

‘Biochemical and genetic characterization of an unusual mild PEX3-related Zellweger spectrum disorder.’

Mol Genet Metab. 121(4):325-328 (2017). doi:10.1016/j.ymgme.2017.06.004.

Falkenberg KD et al.“Allelic Expression Imbalance Promoting a Mutant PEX6 Allele Causes

Zellweger Spectrum Disorder.”Am J Hum Genet. 101(6):965-976 (2017). doi: 10.1016/j.ajhg.2017.11.007.

Falkenberg KD*, Klouwer FCC* et al.

“Autophagy inhibitors do not restore peroxisomal functions in peroxisome biogenesis disorders.”

[manuscript submitted for publication]

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“It takes a VIllage...” – ACknowledgements

This thesis would have never come to its completion without the help of quite a

number of people.

First of all, I’d like to thank my supervisors for their unswerving support. Dear Hans,

thank you for providing me the opportunity to write my PhD thesis in your group.

I could always ask you for advice when struggling, and you helped me re-focus and

re-organize priorities, giving me a shove in the right direction. But most importantly,

you gave me both the chance and the confidence to work rather independently.

Also your stubbornness regarding reviewers’ comments was impressive (and fairly

fruitful), setting a perfect example not to give up too soon. Dear Ronald, your

scientific enthusiasm combined with your encyclopedic knowledge about (probably)

every single metabolic paper ever written was hugely inspirational for me. Thanks for

all your kind words, your open-mindedness and your never-ceasing curiosity.

I consider myself lucky that I could perform my studies in the Laboratory Genetic

Metabolic Diseases, all of its members making it a truly comfortable working

environment. Thanks to each and every one of you for your hospitality and all the

social get-togethers, like lab days, various poules (although I honestly still don’t

entirely get your enthusiasm for the Tour de France) as well as countless gezellige

borrels.

For scientific support, I’d especially like to thank the whole research branch of the

GMZ. Foremost, my gratitude goes to Janet K, who met my countless (and possibly

slightly repetitive) questions with everlasting kindness and the literal patience of a

saint. From day one, I could always count on you, you always made time for me when

I asked for help. Thanks so much! Also without you, Rob O, my thesis would have

definitely lacked quality (and our room wall memorable quotes). Thanks for your

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wonderfully refreshing pragmatic approach to experiments, your huge support was

really a blessing for our whole group.

Dear Carlo, I was inspired by your enthusiasm and your scientific curiosity showing

the heart of a true researcher. I always enjoyed working with you and exchanging

ideas. Dear Lodewijk, thanks for unlimited supply of good advices and especially for

your surprisingly dry humor. You keep the lab (and pipette calibrations) together!

Huge thanks also to Sacha, Petra, Monique, Marjolein and of course the DNA

diagnostic girls, who were always willing to lend experimental help and advice to

me. It was a real pleasure working with you.

I was privileged to not only profit from the resources in the lab GMZ, but to be

also part of the Marie Curie ITN PerFuMe. This brought me in contact with young

researchers with all kind of scientific backgrounds in all over Europe, with the

common goal to decipher the secrets of the best of all organelles. Besides scientific

exchanges, we travelled towns and countries and (by shared experience of progress

meetings, conferences and bachelor parties) we truly developed into so much more

than a just network randomly patched together. I’m so glad I met all of you.

For their hospitality and for everything I learnt during my secondments I’d like to

thank Damien Devos and Nicola, as well Michael Schrader’s lab. Special thanks

to you, Michael, not only for being my official second PerFuMe supervisor but also

for your vast knowledge of immunofluorescence. Thanks (and hugs and kisses) to

Afsoon, for mentoring me during my secondment and for being such a lovable and

cheerful person.

Dear Kathi, you were not only also part of my PerFuMe family, but also my room-

buddy in the office and during conferences. We had a terrific time! Thanks so much

for forgiving me my obvious German-ness, for all the coffee you brought me in just-

too-early meetings, for the experience of sugar-free cake and for being constantly and

reliably there for me throughout the last four years of ups and downs.

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You were also part of our formidable AiO group, to which I want to direct my focus

now. It is said, that a sorrow shared is a sorrow halved, while sharing joy only doubles

it; and in my modest opinion, this was so true for us when sharing the frustrations

and successes of all our projects. I felt that that sharing our research experiences (as

well as piles of cookies) truly bound us together. I knew I could always come to you

to discuss stupid new hypotheses or dinner recipes. And, of course, I will not forget

all our awesome activities, from cake competitions, arts and craft sessions, sunny

lunches, Friday borrels, AiO dinners and Sinterkerst parties up to our trips to the

Lisbon, France and the Efteling (twice!!). Guys, it was truly amazing, you helped me

so much throughout the whole PhD time. Thanks for being there and being you!

Dear Femke, despite your subliminal (word of the day!) OCD, it was a pleasure

working with you and being your next-desk-neighbour. We pretty much hit it

off together, potentially due to sharing a similar kind of (black-ish, occasionally

borderline-offensive) humor. Even on days when I dreaded the thought of going to

the lab, at least I was always looking forward to have a coffee/tea and chat with you.

You made frustrations so much more bearable, and you have this astonishing talent

of cheering me up (mainly by insulting Marte or me). I am so grateful, that we met.

Before risking to get kitschy (and we all know, how much you like that), let me just

say: Thank you so much for everything, honey (particularly also for the legendary

treasure hunt and for agreeing to be my paranymph)!

Dear Marte, although you – as so many in the lab before you – made the unfortunate

choice to turn to the dark side of organelles, I am grateful your companionship in

the last years. How else could I have spread the beauty of German sayings in the

Netherlands? Thanks for your disarming honesty and for being my borrel buddy, my

lovable little soul-ful nerd.

Dear Jia, thanks for your endearing humor and your contagious, almost child-like

excitement and passion for research. Make sure to never lose that. And kudos to you,

girl, you can certainly hold your liquor.

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Dear Clair, thanks for your super-dry humor and your efforts teaching me Dutch

(in merely refusing to talk English with me). Although you were “hesitant” to join us,

you were a welcome princess-like presence in our room, Amsterdams to your very

bones.

Dear Eileen, thanks for your positive spirit, it was always so nice and motivating

talking to you. I admire your aspirations to communicate science to the world and

am really curious where you end up. Research needs people like you!

[To all of these office roomies: Thanks for bearing with my untidiness, slight

unpunctuality, ever-present lab journal complaints and seasonal loops of Christmas

songs. Also thanks to all former office mates, like Kevin, Maxim, Steph, Riekelt and

Naomi, for the great atmosphere and for making it such a fun office to work in.]

Dear Iedan, thanks for your unshaken optimism, all the book recommendations

and your keen interest in science. Despite being the lone bastion of neuroblastoma

research (and the only guy) among the AiOs, you always had an open ear for

everyone’s project, offering valuable input and finding connections to other subjects.

Your organizational skills regarding fairytale themed parks will not be forgotten.

Dear Arwen, your incapability to stop squeaking, when confronted with something

remotely cute, and your crazy science ideas (letting you voluntarily spend weekends

in the lab to test them) make you a true unique in the lab (and probably the world).

Thanks for being so big-hearted and for your awesome moon pies and dumplings.

Dear Jeannette, you granted me asylum when I was homeless. I owe you big thanks

for that, but also for many fun evenings (e.g. the legendary party night with Ntsiki in

Lisbon), for supplying us with fresh fruits and especially for being such a sunny soul.

Dear Ntsiki, you were my coffee buddy, and I will be eternally grateful for the many

times you saved me from coffee-less mornings. With the help of gorgeous flying

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unicorn memes, you always fought yourself out of the worst lab frustrations, and still

had time to listen to my newest “hypothesis”. Thanks for making us have our fantastic

ski trip in France, and for doing your best to motivate us for yoga camps, sunny

lunches and summer schedules.

Dear Suzan, although you seem so quiet and innocent on first sight, we all know by

now about your dark-ish kind of humor accompanied by your adorable dirty laugh. I

admire the stamina you proved when faced with the sorrows of lab work. Thanks for

always having a smile for everyone!

My dear Iliana, of course, I did not forget you. Together we fought through late

evenings in the lab and failed experiments, just like through crowds at all the bars,

concerts, festivals and parades we visited together. I am so grateful for all the time

we spent together (despite the fact that you might be the slowest eater and drinker in

history of humankind). Thanks for being such a laid-back and charming person, and

especially for being always a tiny bit more delayed than I am.

Despite working only with you for a shorter while, I’d like to thank Kevin for all

his help, jokes and peroxisomal enthusiasm, Marco Moedas for the Lisbon trip and

valuable life lessons (i.e. always carry cash), Olga M for sophisticated book talks and

the most beautiful articulation, and Sanne for her quirky sense of humor.

Moreover, I’d like to thank Marco for all the hugs offered when needed and for giggling

through the lab with Marte like two 4 year-old besties. Thanks to you, Rashmi, for

your warm and calm presence, and always having a nice word for everyone. Also

thanks to former members of the lab, Anita, Sandra, Olga P, Eveline, Eugen and

Martin, for your help and all your good advises.

I especially also like to thank you, Riekelt, for your constant interest in my projects,

your help with my application, and for your willingness to improve your German

pronunciation. To you and Naomi, Reuben, Alyson, Rob W, Michel and Fred I also

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owe many enlightening conversations during borrels. Special thanks to Michel for

clearing out my desk and taking care of the kids.

Apart from all those people helping with my research, I would also like to express my

gratitude to the people who kept me sane(-ish) during the last years.

Meine lieben Couch Potatoes, da ich den Eindruck hatte, dass ihr mein Projekt leicht

missverstanden habt (nämlich der festen Meinung seid, dass ich an gefährlichen

Viren arbeite mit dem erklärten Ziel, die Weltherrschaft zu ergreifen), hoffe ich

nun, dass euch diese Doktorarbeit vom Gegenteil überzeugt. Lieben Dank euch

für Bootstouren, Strandtrips und zahllose Dinner- und Bar-Abende. Toll, dass ich

zusammen mit euch Amsterdam entdecken durfte. Mit dem Niederländisch hat es

dann ja auch fast geklappt. Danke für alles!

Auch dicken Dank an Laura, Anna und Anne, dafür dass ihr mich so perfekt vom

Alltag abgelenkt hat. Mit euch ist es immer noch genauso lustig und ungezwungen,

als hätten wir gestern noch die Schulbank gedrückt. Die Reisen nach Rom, Kanada

und Australien waren einfach toll!

Zu guter Letzt möchte ich meiner Familie danken. Ich fürchte ein wenig, dass diese

Zeilen schwerlich daran herangekommen, wie dankbar ich euch allen tatsächlich

bin, aber ich werde mein Bestes tun. Ihr seid immer und unerschütterlich für mich

da, und zeigt sogar Interesse für meine Projekte, auch wenn es noch so nerdig wird.

Danke Oma, dass du mich immer mit offenen Armen empfängst (und natürlich

für die Riesenladung Spritzgebäck jeden Winter). Danke Linda (meiner Einhorn-

Paranymphe) und Christian, dass ihr in meinem Leben seid, dass ihr mich zum

Lachen bringt, für die Hilfe beim Cover (auch wenn blau-gelben Punkte letztendlich

doch das Nachsehen hatten), und vor allem für Oscar, mit dem die Welt doch noch

tausend mal so bunt und sonnig scheint.

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Besonderer Dank geht natürlich an meine Eltern. Liebe Mama, lieber Papa, danke für

eure ehrlichen Kritiken und Ratschläge, für eure uneingeschränkte Unterstützung in

allen Lebenslagen (und bei zahlreichen Umzügen) und für die Gewissheit, dass ich

mich hundertprozentig auf euch verlassen kann (und egal was ihr sagt, das ist bei

Weitem nicht selbstverständlich). Ihr seid für mich wie ein sicherer Hafen, in den ich

immer zurückkehren kann, wie unruhig es sonst auch ist. Ich bin unglaublich stolz

und glücklich, euch alle zu haben. Ich hab euch sehr lieb.

Knowing myself, there is a pretty high chance that some people, which I owe my

gratitude to, slipped my mind (including for instance quite a number of students).

For all those forgotten: Thanks a lot, guys (and my sincere apologies)!!

Out of respect and a healthy portion of self-preservation, I decided to refrain from

adding any photographic memoirs to this section. But be not be mistaken, my friends,

they will all be stored at a safe place.

Till then, I wish you farewell and all the best for your future.

Thanks for having joint the ride, it’s been a blast.

Yours,

Kim