Nederlandse Samenvatting Appendices 175

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1 UvA-DARE (Digital Academic Repository) Blood coagulation-(in)dependent protease activated receptor signaling in pancreatic cancer Shi, K. Link to publication Citation for published version (APA): Shi, K. (2015). Blood coagulation-(in)dependent protease activated receptor signaling in pancreatic cancer General rights It is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulations If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. UvA-DARE is a service provided by the library of the University of Amsterdam ( Download date: 14 Feb 2017

2 Nederlandse Samenvatting Appendices 175

3 Nederlandse Samenvatting Nederlandse Samenvatting Alvleesklierkanker is zonder twijfel een van de meest dodelijke ziektes, met alleen marginale mogelijkheden tot behandeling. PARs (protease activatedreceptors) zijn receptoren die verantwoordelijk zijn voor de cellulaire processen geïnduceerd door bloedstollingsfactoren. PARs blijken echter ook een rol te spelen bij het verloop van verschillende ziektes, waaronder mogelijk ook kanker. In hoofdstuk 1 wordt een introductie geven in de bloedstollingscascade, alsook de PAR signalering en de mogelijke relatie tussen PAR en kanker. In hoofstuk 2 proberen we de rol van PAR2 in de groei en migratie van alvleesklierkankercellen te ontrafelen. Het blijkt namelijk dat PAR2 sterk tot expressie komt in de kankercellen van alvleesklierkankerpatiënten, alsook in alvleesklierkankercellijnen. Dit in tegenstelling tot gezonde alvleeskliercellen waar het PAR2 niveau laag in is. We tonen aan dat PAR2 geen effect heeft op de groei van alvleeskliercellen, maar wel op de migratie ervan. Opmerkelijk genoeg komt de migratie alleen voor in een ATP rijke omgeving. Dit kan van belang zijn omdat juist in tumoren hoge ATP concentraties voorkomen, en omdat behandeling leidt tot een toename van ATP in necrotisch tumorcellen. Dit kan betekenen dat PAR2 gedreven migratie van kankercellen plaatsvindt in patiënten met tumornecrose en/of bij kankerceldood door antikanker behandeling. In hoofdstuk 3 tonen we aan dat PAR2 niet alleen tot expressie komt in kankercellen, maar ook in niet tumorigene cellen in de tumor, het zogenaame tumor micromilieu. Met behulp van een PAR2 knock-out muis model tonen we aan dat PAR2 primaire tumorgroei remt, maar tegelijkertijd wel metastase via de lymfe induceert. Opmerkelijk genoeg zetten alvleesklierkankercellen epitheelcellen afkomstig van de lymfevaten aan tot het vormen van bloedvatstructuren, en wordt dit effect juist weer geremd door PAR2. Hiermee tonen we dus een nieuwe rol van PAR2 aan in 176

4 Nederlandse Samenvatting tumorgroei, namelijk door de remming van de groei van lymfatisch vaatweefsel (lymfangiogenese). PAR2 speelt daarmee een opmerkelijke dubbel rol in de ontwikkeling van alvleesklierkanker, door de inductie van primaire tumorgroei aan de ene kant, en remming van metastate via de lymfevaten aan de andere kant. Behandeling van kankerpatiënten met PAR2 remmers is daarom niet vanzelfsprekend, en vooral niet in het geval van alvleesklierkanker. In hoofdstuk 4 proberen we te ontdekken hoe PAR2 de lymfangiogenese kan remmen. Als eerste laten we zien dat remming van lymfangiogenese door PAR2 specifiek is voor alvleesklierkanker, aangezien het niet wordt waargenomen in andere soorten kanker. Vervolgens zijn 10 eiwitten geïdentificeerd die potentieel verantwoordelijk zijn voor de remming van lymfangiogenese door PAR2. Van twee van deze eiwitten, PDGFA en TAC1, is al bekend dat ze lymfangiogenese zouden kunnenbeïnvloeden en zijn daarom extra interessant. Momenteel proberen we te achterhalen of een van deze 10 eiwitten ook daadwerkelijk lymfangiogenese remt. Het identificeren van een dergelijke remmer is niet alleen belangrijk voor de behandeling van alvleesklierkankerpatiënten, maar ook om een beter begrip van lymfangiogenese te krijgen, wat de behandeling van kanker in het algemeen ten goede kan komen. Vervolgens richten we ons op PAR1, een eiwit dat hoog tot expressie komt in verschillende soorten kanker en gelinkt is aan de ontwikkeling van de ziekte alsook de genezing. In hoofdstuk 5 tonen we aan dat PAR1 overal aanwezig is in het tumor micromilieu. Met een vergelijkbare methode zoals gebruikt in hoofdstuk 3 wordt aangetoond dat PAR1 de progressie van alvleesklierkanker bevordert. Door de expressie van PAR1 worden verder macrofagen aangetrokken in de tumor, wat leidt tot resistentie van de tumor tegen de chemische bestrijding (chemokuur). Dit duidt mogelijk op een niet eerder geïdentificeerde rol van PAR1, en inhibitie van PAR1 zou de efficiëntie van een chemokuur behandeling kunnen verhogen. 177

5 Nederlandse Samenvatting Zoals de naam al aangeeft worden PARs geactiveerd door proteases, en een bekende activator van PAR1 is trombine. In hoofdstuk 6 richten we ons daarom op trombine, door te kijken naar het effect van de trombineremmer dabigatran op de groei van alvleesklierkanker, alsook het ontstaan van mogelijke chemoresistentie. Wanneer dabigatran alleen wordt toegediend, heeft het geen effect op tumorgroei. In combinatie kan het echter de werking van gemcitabine als kanker remmerverhogen. Dabigatran heeft vreemdgenoeg geen effect op het aantrekken van macrofagen door de tumor, noch op de desmoplastische reactie karakteristiek voor chemoresistente pancreastumoren. Onze hypothese is dat dabigatran-afhankelijke bloedvatvorming de groei van tumorcellen bevorderd, waardoor deze gevoeliger worden voor gemcitabine. In muizen versterkt dabigatran de remming van alvleesklierkanker door gemcitabine, echter in mindere mate dan PAR1. Het werk in dit proefschrift toont aan dat PAR1 and PAR2 een belangrijke rol spelen in de ontwikkeling van alvleesklierkanker. Het remmen van PAR1 kan relevant zijn bij de behandeling van patiënten, hoewel rekening gehouden moet worden met het frequent ontstaan van bloedingen. De onverwachte dubbelrol van PAR2 in alvleesklierkanker suggereertechter dat remming van dit eiwit niet de meeste logische behandelingsmethode zal zijn. In plaats hiervan zou het relevanter zijn om de werking van de, nog niet geïdentificeerde, remmer van PAR2-gestuurde lymphangiogenesis te verhogen. Ondanks dat bloedstolling bijdraagt aan de ontwikkeling van alvleesklierkanker, heeft behandeling met bloedstollingremmers niet geleid tot positieve effecten. De dubbelrol van PAR2 zou hier mogelijk een verklaring voor kunnen geven. Bloedstollingremmers remmen namelijk PAR2 activering door TF/FVIIa of FXa, wat uiteindelijk leidt tot toename van metastase via de lymfe. Dit effect kan daarmee de voordelen van remming van PAR1 (door remming van trombine) overschaduwen. 178

6 Chinese Summary/ 摘要 摘要 (Chinese Summary) 毋容质疑, 胰腺癌已经成为人类最致命的癌症之一, 而目前胰腺癌治疗方案也十分有限 在传统认知中, 蛋白酶激活受体 (protease activated receptor, PAR) 被血凝因子激活, 在血凝结过程中起到重要作用 现如今, 蛋白酶激活受体在癌症发展生理学的重要位置业已被普遍认可 在本书第一章中, 我们首先介绍了血液凝结的机理 蛋白酶激活受体的信息传导方式以及两者与癌细胞发展的关联性 在第二章中, 我们着重于阐明蛋白酶激活受体 2(PAR-2) 在胰腺癌细胞中对细胞繁殖和转移的重要影响 我们发现 PAR-2 在胰腺癌细胞和病人的肿瘤样本中过量表达, 然而却几乎不会在健康的胰管中被检测出来 我们同时发现, 胰腺癌细胞上被激活的 PAR-2 并不会增强癌细胞的繁殖能力, 但是会大大增强细胞的移动能力 这样显著增强的移动能力同时也需要细胞外 ATP 的参与 更重要的是, 在癌细胞中 ATP 的含量要远远高于健康细胞中 ATP 的释放量, 而往往一些传统上的治疗方式会诱导坏死的癌细胞大量释放 ATP 我们的数据显示 PAR-2 引导的胰腺癌细胞转移有着广泛的临床影响 在第三章中, 我们展示了 PAR-2 并不仅仅被胰腺癌细胞表达, 同时也广泛的存在于肿瘤微环境中 我们使用了原位癌细胞移植的小鼠模型, 将癌细胞移植到小鼠的胰腺中, 观察癌细胞在免疫系统正常环境中的发展 我们发现当小鼠的 PAR-2 被遗传切除后, 也就是说, 只有癌细胞表达 PAR-2 而肿瘤基质区室以及周围环境并不表达 PAR-2 的情况下, 原位肿瘤的生长得到了控制, 但是癌细胞转移到淋巴结的能力却得到了增强 有趣的是, 在血管成型实验中, 胰腺癌细胞可以引导淋巴管内皮细胞形成管状结构, 但是当淋巴管内皮细胞上的 PAR-2 被激活后, 由胰腺癌细胞引导的成管能力反而被抑制 我们的数据还证实了 PAR-2 在肿瘤微环境中的新功能以及 PAR-2 对于淋巴导管生成 179

7 Chinese Summary/ 摘要 的抑制作用 基于我们的观察发现,PAR-2 显然在胰腺癌肿瘤微环境中起着双重作用 一方面,PAR-2 可以辅助原位癌细胞的增长, 另一方面,PAR-2 的存在却会抑制淋巴导管的生成, 继而降低淋巴转移的风险 由此可见, 虽然抑制 PAR-2 在其他癌症上也许有明显作用, 但是在胰腺癌的临床应用上, 倘若要使用 PAR-2 抑制剂, 应该尤其注意抑制后的负面影响 在第三章中, 我们并没有明确 PAR-2 引导的淋巴习惯生成抑制的机理 我们在第四章中对这个抑制机理做了进一步的研究, 并希望能够最终确认相关的抑制剂 我们首先展示 PAR-2 引导的淋巴管生成抑制只存在于胰腺癌中, 在其他用来测试的癌细胞种类中 ( 例如乳腺癌 大肠癌等 ),PAR-2 并没有这个作用 而后, 我们使用了基因分析技术, 并且最终确定了 10 个可能相关的蛋白质, 并把这 10 个蛋白质作为进一步的研究对象 在这 10 个蛋白中, 至少有两个 (PDGFA 或者 TAC1) 曾被认为与淋巴管的生成有一定的联系 与此同时, 我们正在致力于确定哪一个才是最终的抑制剂 很明显, 淋巴管生成抑制剂的确定, 将会对胰腺癌的治疗有很大的帮助, 并且能够让我们在淋巴管生成与 PAR-2 的关系上获得更深的认识 对于淋巴管生成机制的进一步了解, 也将会对其他癌症治疗提供有用的帮助 在第五章和第六章中, 我们也对 PAR-1 进行了研究 多方研究曾证明,PAR-1 在不同的癌细胞种类中高度表达, 并且在一些特定的癌症病例中,PAR-1 的高度表达和该疾病的发展以及病人的生存状况有着明显的联系 在第五章中, 我们首先展示了 PAR-1 遍布于胰腺癌的肿瘤微环境中 然后我们使用了第三章中提到的小鼠模型, 对其进行了 PAR-1 基因切除 实验证明,PAR-1 的存在会直接促进癌细胞的发展, 以及更重要的是, 胰腺癌肿瘤微环境中 PAR-1 通过巨噬细胞直接导致了癌细胞的抗药性 我们的实验数据显示了 PAR-1 在肿瘤微环境中的全新作用, 同时也证明了 PAR-1 的辅助功效在普通治疗中亦尤为突出 如第一章中提到的,PAR-1 会被不同的酶激活, 其中凝血酶 180

8 Chinese Summary/ 摘要 (thrombin) 是 PAR-1 最典型的激活剂 在第六章中, 我们对凝血酶直接抑制剂 Dabigatran 在胰腺癌的生长和抗药性上的作用进行了深入研究 在我们的动物实验中,Dabigatran 作为单一疗法并没有影响原发肿瘤的生长速度, 但是当加入化疗药物 gemcitabine 的时候,Dabigatran 明显增强了 gemcitabine 的抗癌效果 有趣的是,Dabigatran 并没有通过巨噬细胞来完成这一反应 由此我们得出结论, 依赖于 Dabigatran 而促进的血管生成, 使原位肿瘤增长加速, 从而使得 gemcitabine 更为有效 总而言之,Dabigatran 增强了胰腺癌细胞对 gemcitabine 的敏感度, 虽然这一效果并不如 PAR-1 基因完全切除那么明显 在这部论文中, 我们着重阐明了 PARs 在胰腺癌发展中的关键作用 抑制 PAR-1 对于临床应用有着重要意义 然而抑制 PAR-1 后带来的副作用 ( 比如出血 ) 也不可忽视 基于 PAR-2 在胰腺癌肿瘤微环境中的双重作用, 我们强烈建议, 对于胰腺癌患者来说, 抑制 PAR-2 并不是一个明智的选择 更为重要是, 须首先明确 PAR-2 引起的淋巴管生长抑制的抑制剂 PAR-2 在胰腺癌肿瘤微环境上的双重作用同时也揭示了为什么在实际临床中抗血凝药物对胰腺癌病人的作用微乎其微 尽管血液凝结对癌细胞的发展有着不可或缺的影响和联系, 通常情况下, 一个抗血凝药物并不仅仅影响血凝途径中的一个因子, 而是会影响到多个因子, 从而使过程变得更为复杂, 结果也会更加不可预料 ( 本文内容以英文为准, 中文摘要仅作参考 ) 181

9 Dankwoord Dankwoord/Acknowledgments/ 致谢 We all owe everyone for everything that happens in our lives. But it s not owing like a debt to one person-it s really that we owe everyone for everything. Our whole lives can change in an instant-so each person who keeps that from happening, no matter how small a role they play, is also responsible for all of it. Just by giving friendship and love, you keep the people around you from giving up- and each expression of friendship or love may be the one that makes all the difference. 我们生活中发生的每一件事, 都受到其他人的恩惠 但这并非象是负债于某人, 我们拥有的一切都应该归功于每一个人 人生可能会在一瞬间改变, 所以每一个让你人生步入正轨 稳步向前的人, 都值得感激, 无论他们扮演的角色多么卑微 只要给予他人友谊与爱, 你身边的人就不会轻言放弃 任何一个友谊与爱的表达都能让一切变得美好 <the end of your life book club >by Will Schwalbe This is almost at the end of the tenth year since I arrived in the Netherlands. It has never been easy to stay in a foreign country even after so many years. I think I am just lucky to have all of you. With all your support, I reach to here. I decided not to try to list all you below only because I was too worried about accidentally leaving someone out; but I am grateful beyond words to all of you. (Please allow all those grammar mistakes, I cannot ask someone else to check the grammar for this part :P) I must give my first word to my parents and my grandparents. Without your unconditional love and unlimited financial support, nothing would ever happen. You winged my dream. Unlike other ordinary Chinese parents, you respect all the choices that I have made so far, and fully encourage and support me to do everything I want to do. I believe, it was, and still is difficult to let me go so far away from you since I am the only child. But you kept the struggles and worries within yourselves while leaving me free. I would like to give my special thanks to my grandfathers. You both offers/ed me a rich spiritual life, and such spiritual influences implanted into my soul, and they are priceless gift that I will bring with me for the entire life. ( 我必须要把第一个感谢的位置留给我的父母和姥姥爷爷 没有你们无条件的爱与经济支持, 我就不会有现在 你们为我的梦想插上翅膀, 并且让我自由的去飞翔 不同于一般的中国父母, 你们让我决定自己想要走的路, 并且全力的支持我所走的每一步 我相信无论过去还是现在, 两个国家的距离对于你们来说都是艰难的选择和决定, 但是 182

10 Dankwoord 你们把担心留给了自己, 并且选择放手给我自由 另外, 我特别要感谢爷爷 你们给了我富裕的精神生活, 这样的精神影响是无价的, 它们根深蒂固在我的性格中, 也将一路伴随我走的更远 ) Arnold, you are one of the best and most important steps that I have made in my life. Everyone knows how important a good supervisor meant for a PhD student, and you are EXACTLY the right example. Your knowledge and your patient lightened my way. You always available when I am having questions and you always willing to help me in order to reach to the final destination. There is not enough word to describe how much I feel appreciated, but I am truly feeling lucky to have you as my supervisor for all this period. Dick, my promoter. Thank you for all the useful advices from doctor s perspective, and you keep me updated with things happening in the clinical world. Especially thank you for your effort and support in the last period during my preparing for the defense. I would also like to thank the committee members for evaluating this thesis, as well as challenging me during the defense. Maaike, thank you to be my paranymph. It probably was a bit surprise to you that I asked you to be my paranymph, but I truly enjoy your type of humor and I also thank you for all the helps you offered when I needed a hand. A lot of thanks to the stolling group - a small but cozy group. Hella, Lin, JanWillem, thank you that made my working life so beautiful and enjoyable apart from all the scientific and technique support. Brendon, thanks for all those bioinformatics helps. Those fancy bioinformatics graphics made my chapter much more pretty. People sat/sit in L Thank you all for your gezelligheid. Special thanks to Karla, I appreciate a lot of your help and knowledge on pancreatic cancer research. I enjoyed very much the moment we went for dinner in Bergamo and all those nice talks we had. It was pity that you left the group so early, but I believe you are extremely enjoying the nice sunshine in Brazil with your newborn baby. I d like also thank Arjan who developed the scratch analysis program on ImageJ, you really saved my life. Otherwise, I may collapse in front of my PC while analysis the scratch assay 183

11 Dankwoord one by one. Kaushal, it was a very enjoyable moment when you were sitting in the same room and especially we were sharing the table. You are very organized and highly motivated, and same time you are also very knowledgeable and smart. I wish you will go very very far on what you like to do and I am sure you definitely deserve a big success. Luigi, you are just special in many ways. I sometimes amazed by how much energy and enthusiasm you are donating in scientific world. You are smart and your sincere attitude toward to science convinces me that you will reach to your dreamed success. Apart from work, I really enjoyed your inspiring talks and your hospitality. It is also pleasure to discover your passion on food qualities. I wish you a lot of success in your next steps. Danielle S, I enjoy your fashion style very much <3. Tim, thanks a lot for all the nice talks. Daan, especially thank you for the help during the moment when I had to communicate with the terrible AMC postkantoor about my missing package. Joost&Maarike, you both are awesome. I cannot imagine doing animal experiments without you. Because of you, everything went so smooth and well, your effort is highly appreciated. Meanwhile, deeply appreciation to those mice whom donated their life for my experiments. Your inputs meant a lot to me. As a Buddhist, we speak about Karma, and I believe your next life will be ungraded in a higher cycle. As a human being, I thank for your donation in making human life better. Monique, every time when I need tracking an order or when I just simply need something, you are always helpful. And for those moment you were on holiday, I felt pretty insecure about making an order. Heleen and Monique (the secretory), thank you for make my life much easier in many ways when I had to dealing with all those documents and forms. Helene, it was a fantastic California trip with you. It s pity that we could not do it much longer. Additionally, thank you to share your knowledge, critical views and passion on science with me. I was very glad that you worked during the same period as I did. I wish you a lot of good luck in the future and more moment that is enjoyable in life. My great grateful to Adrian Begg. It is sad that you will never read this anymore, but 184

12 Dankwoord that is not the reason to hide my appreciation away. You are not only opening the door of scientific world to me, you also shown me how much you loved and enjoyed your life after work. I always believe a healthy private life is much more important than any success on career. Your type of humor was very special and your kindness was distinct. God needed a better person next to him, which was maybe the reason that he took you away. Peter S, thank you so much for all the helps on making lentivirus, and thank you for your socialness and kindness. Although I haven t had a chance to put the lentivirus paper in this thesis, it s coming!!!! And thank you for being so patient when I asked some stupid questions. Arjan L, thank you for your help in arranging the difficult ordering with me. In that way, we really saved a lot of money :D. Meanwhile, I also enjoy your photograph very much, especially those ones you shared on FB. Please keep on going!! It is always nice to see the beauty of the world where I haven t been yet. To all the friends I meet/met in the NL (so many names, that I am not gonna list everyone here, but you know I am talking about you :D ), and specially to all R s friends (Erik, Douwe, Helen, Bernard, Klaas,. etc.). I know I am not an ordinary social person, and I am sometimes boring like a hell. However, it never stops me away from enjoying spending time with all of you, such joyful time riches my life in the Netherlands. 慧慧, 谢谢你在如此 关键 的时候和我分享那么多心得 也感谢你曾经在 Wageningen 的时候帮助我的一切 Nona, 煽情的话就不说了, 你要过的快乐 <3. D, 感谢你无条件为我所做的一切, 我希望你的生活能幸福平安 阳, 私心觉得你该好好给我写个感谢信啊哦哈哈哈 但是依旧很感谢你这么多年来, 一直都在那里 不同于一般西方人眼中的中国人, 你向 R 展示了你的独立和冒险精神 太多要夸你的了, 不知道该哪个词 (Adventure, open-mind, independent, humor and easy-going.) 希望你能继续保持下去 Fay. 从 12 岁, 到现在, 感谢一路走来, 有你在 人生长长短短, 宇宙无限 渺小如我, 却也可以在聚散离合的道路上与你一起向前 成长在路上, 感谢你与我一同成长, 在同一条路上, 直到生命终止的那一刻 Additionally, thank you for 185

13 Dankwoord designing the cover and the bookmark of this thesis, that s an amazing piece of art. 我亲爱的妞妞, 好像还没有在这样的场合说过什么, 很多话都只可意会不可言传, 所以其实这一段也变得蛮难写 不过我们在荷兰相依为命的日子还很长 感谢在遥远的异国他乡还能有家人在身旁, 让我们可以分享的不只是童年还有成长和未来 Unlimited appreciation and love to my cats: Kiki and Gray. Although both of you will never understand what I am saying, you will get special food as gift to thank for the contribution, tolerance and cuteness that you give me. Dear Rutger, I do not deserve someone as wonderful as you, and you sure don t deserve someone as trying as me. But I am insanely lucky and you are incredibly positive and patient. You went through with me all my darkest moment with unconditional love and support. You are the light of my life. A quote from a famous Chinese writer describe the best: 我走过许多地方的路, 行过许多地方的桥, 看过许多次数的云, 喝过许多种类的酒, 却只爱过一个正当最好年龄的人. Finally, to the little one, thank you for joining my life during such special moment, you made this period extra spectacular. 要感谢的人有很多, 不只是这四年来, 而是这 20 多年来对我有过帮助有过激励有过影响的所有人 只是我无法把你们一一列出, 所以谨此一句 谢谢, 感谢那些在我生命中出现过的有名的, 无名的人, 相信你们会明白 我在我的一席之地上闪耀, 希望你 ( 们 ) 也是 186

14 Curriculum vitae Curriculum vitae/self-introduction I was born on the 18 th July 1985 in Beijing, China. I spent my wonderful childhood mostly with my grandparents. I never was an ordinary Alfastudent in the class because I simply did not like to be the way in which Chinese teachers wanted me to be, I was too curious about many things that irrelevant to what I learnt from the school. Therefore, after a routine school education, I decided to go outside and visit the world. I was hesitating to make the choice between fashion design and biotechnology at the beginning, but the magic of science easily caught my attention right after. I therefore started Bachelor of Biotechnology in Wageningen University in As one of the few foreigners in the class at that time, the experiences were a mix of special, exciting, difficult, distance and thinking. I moved to Amsterdam and live there in 2007 while I was still studying in Wageningen, and traveling between two cities was my daily activity last until the end of my study. Time flew fast, at the end of my bachelor study, I discovered that I have more and more interests toward to molecular level cancer research. Not only because it sounds cooler, but also reveals me an indefinite possibilities on seeking the novel features. In 2008, I started my master on Molecular sciences with the division on biomedical research in Wageningen University. I accomplished my Master thesis on DNA repair at the Netherlands Kanker Institute (NKI) at the department experimental therapy under the supervision of Conchita Vens and Adrian Begg. That was one of my most grateful period in my scientific life. Afterwards, I finished an internship at Oncogenetics department at Academic Medical Center in 2010 and started my PhD at the department Center of experimental and molecular medicine in August in the same year. I am now working as a post-doc and try to make even further steps on the subject of pancreatic cancer and blood coagulation, and I believe I will start exploring other possibilities in the near future. 187

15 Portfolio Proefschrift Portfolio Proefschrift Name PhD student: Kun Shi PhD period: August 2014-Febury 2015 Name PhD promoter: Prof. dr. Dick. J. Richel Name PhD co-promoter: Dr. C. Arnold Spek. PhD Training Course Year Hours/ECTs The AMC World of Science /0.7 Cardiovascular Diseases-Thrombosis and Hemostasis /1.5 Laboratory Animals /3.9 Scientific Writing in English for Publication /1.5 Anatomy of the House Mouse /1.1 BROK /0.9 Entrepreneurship in Health and Life Sciences /1.1 Seminar, workshops and master classes Department Seminars AMC oncology Seminars Presentations and conferences International conference on thrombosis and hemostasis issue in cancer (ICTHIC) (poster presentation) /0.9 Tumor Cell Biology meeting (KWF) 2011 / /0.7 Pancreatic cancer meeting (AACR) (twice poster presentation) International Society on Thrombosis and Haemostasis (ISTH) (oral presentation) / /

16 List of Publications List of Publications (so far) 1. Protease-activated receptor-2 induces migration of pancreatic cancer cells in an extracellular ATP-dependent manner. Kun Shi, Karla Cs Queiroz, Jan Stap, Dick J Richel, C Arnold Spek Journal of Thrombosis and Haemostasis. 2013;11(10): Protease-Activated Receptor 2 Suppresses Lymphangiogenesis and Subsequent Lymph Node Metastasis in a Murine Pancreatic Cancer Model. Kun Shi, Karla C.S. Queiroz, Joris J.T.H. Roelofs, Carel J.M. van Noesel, Dirk J. Richel, C. Arnold Spek Journal of Pathology. 2014;234(3): May protease activated receptor-2 antagonists have clinical potential? Kun Shi and C. Arnold Spek Journal of Pathology. 12/2014; DOI: /path Protease-activated receptor-1 drives pancreatic cancer progression and chemoresistance by promoting tumor associated macrophage recruitment Karla CS Queiroz, Kun Shi, Janwillem Duitman, Hella L Aberson, Johanna W Wilmink, Carel Jm van Noesel, Dick J Richel, C Arnold Spek International Journal of Cancer. 2014; 15;135(10): Protease activated receptor-1 deficiency diminishes bleomycin-induced skin fibrosis. Janwillem Duitman, Roberta R Ruela-de-Sousa, Kun Shi, Onno J De Boer, Keren S Borensztajn, Sandrine Florquin, Maikel P Peppelenbosch, C Arnold Spek Molecular medicine 2014;18;20: Thrombomodulin is a determinant of metastasis through a mechanism linked to the thrombin binding domain but not the lectin-like domain. Netanel A Horowitz, Elizabeth A Blevins, Whitney M Miller, Ashley R Perry, Kathryn E Talmage, Eric S Mullins, Matthew J Flick, Karla C S Queiroz, Kun Shi, C Arnold Spek, Edward M Conway, Brett P Monia, Hartmut Weiler, Jay L Degen, Joseph S Palumbo Blood 07/2011; 118(10):