@phdthesis{Hagedorn2011,
  author    = {Hagedorn, Ina},
  title     = {Novel mechanisms underlying arterial thrombus formation: in vivo studies in (genetically modified) mice},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-85752},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2011},
  abstract  = {Thrombus formation at sites of vascular lesions is a dynamic process that requires a defined series of molecular events including the action of platelet adhesion/activation receptors, intracellular signal transduction, cytoskeletal rearrangements and activation of plasma coagulation factors. This process is essential to limit post-traumatic blood loss but may also contribute to acute thrombotic diseases such as myocardial infarction and stroke. With the help of genetically modified mice and the use of specific protein inhibitors and receptordepleting antibodies, the work presented in this thesis identified novel mechanisms underlying thrombus formation in hemostasis and thrombosis. In the first part of the study, it was shown that von Willebrand Factor (vWF) binding to glycoprotein (GP)Iba is critical for the formation of stable pathological thrombi at high shear rates, suggesting GPIba as an attractive pharmacological target for antithrombotic therapy. The subsequent analysis of recently generated phospholipase (PL)D1-deficient mice identified this enzyme, whose role in platelet function had been largely unknown, as a potential target protein downstream of GPIba. This was based on the finding that PLD1- deficient mice displayed severely defective GPIba-dependent thrombus stabilization under high shear conditions in vitro and in vivo without affecting normal hemostasis. The second part of the thesis characterizes the functional relevance of the immunoreceptor tyrosine-based activation motif (ITAM)-bearing collagen receptor GPVI and the recently identified hemITAM-coupled C-type lectin-like receptor 2 (CLEC-2) for in vivo thrombus formation. Genetic- and antibody-induced GPVI deficiency was found to similarly protect mice from arterial vessel occlusion in three different thrombosis models. These results confirmed GPVI as a promising antithrombotic target and revealed that antibody-treatment had no obvious off-target effects on platelet function. Similarly, immunodepletion of CLEC-2 by treating mice with the specific antibody INU1 resulted in markedly impaired thrombus growth and stabilization under flow in vitro and in vivo. Furthermore, it could be demonstrated that double-immunodepletion of GPVI and CLEC-2 resulted in severely decreased arterial thrombus formation accompanied by dramatically prolonged bleeding times. These data revealed an unexpected redundant function of the two receptors for in vivo thrombus formation and might have important implications for the potential development of anti-GPVI and anti-CLEC-2 antithrombotic agents. The third part of the thesis provides the first functional analysis of megakaryocyte- and platelet-specific RhoA knockout mice. RhoA-deficient mice displayed a defined signaling defect in platelet activation, leading to a profound protection from arterial thrombosis andand ischemic brain infarction, but at the same time also strongly increased bleeding times. These findings identified the GTPase as an important player for thrombus formation in hemostasis and thrombosis. Based on the previous proposal that the coagulation factor (F)XII might represent an ideal target for safe antithrombotic therapy without causing bleeding side effects, the last part of this thesis assesses the antithrombotic potential of the newly generated FXIIa inhibitor rHAInfestin- 4. It was found that rHA-Infestin-4 injection into mice resulted in virtually abolished arterial thrombus formation but no change in bleeding times. Moreover, rHA-Infestin-4 was similarly efficient in a murine model of ischemic stroke, suggesting that the inhibitor might be a promising agent for effective and safe therapy of cardio- and cerebrovascular diseases.},
  subject      = {Thrombus},
  language  = {en}
}
@phdthesis{Emmert2023,
  author    = {Emmert, Martin},
  title     = {The Influence of Substrate Micro- and Nanotopographies on Essential Cell Functions},
  doi       = {10.25972/OPUS-32779},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-327796},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {The introduction of novel bioactive materials to manipulate living cell behavior is a crucial topic for biomedical research and tissue engineering. Biomaterials or surface patterns that boost specific cell functions can enable innovative new products in cell culture and diagnostics. This study aims at investigating the interaction of living cells with microstructured, nanostructured and nanoporous material surfaces in order to identify distinct systematics in cell-material interplay. For this purpose, three different studies were carried out and yielded individual effects on different cell functions. Cell migration processes are controlled by sensitive interaction with external cues such as topographic structures of the cell's environment. The first part of this study presents systematically controlled assays to investigate the effects of spatial density and local geometry of micron scale topographic cues on amoeboid migration of Dictyostelium discoideum cells in quasi-3D pillar fields with systematic variation of inter-pillar distance and pillar lattice geometry. We can extract motility parameters in order to elucidate the details of amoeboid migration mechanisms and consolidate them in a two-state contact-controlled motility model, distinguishing directed and random phases. Specifically, we find that directed pillar-to-pillar runs are found preferably in high pillar density regions, and cells in directed motion states sense pillars as attractive topographic stimuli. In contrast, cell motion in random probing states is inhibited by high pillar density, where pillars act as obstacles for cell motion. In a gradient spatial density, these mechanisms lead to topographic guidance of cells, with a general trend towards a regime of inter-pillar spacing close to the cell diameter. In locally anisotropic pillar environments, cell migration is often found to be damped due to competing attraction by different pillars in close proximity and due to lack of other potential stimuli in the vicinity of the cell. Further, we demonstrate topographic cell guidance reflecting the lattice geometry of the quasi-3D environment by distinct preferences in migration direction. We further investigate amoeboid single-cell migration on intrinsically nano-structured, biodegradable silica fibers in comparison to chemically equivalent plain glass surfaces. Cell migration trajectories are classified into directed runs and quasi-random migration by a local mean squared displacement (LMSD) analysis. We find that directed movement on silica fibers is enhanced in a significant manner by the fibers' nanoscale surface-patterns. Further, cell adhesion on the silica fibers is a microtubule-mediated process. Cells lacking microtubules detach from the fibers, but adhere well to glass surfaces. Knock-out mutants of myosin II migrating on the fibers are as active as cells with active myosin II, while the migration of the knock-out mutants is hindered on plain glass. We investigate the influence of the intrinsically nano-patterned surface of nanoporous glass membranes on the behavior of mammalian cells. Three different cell lines and primary human mesenchymal stem cells (hMSCs) proliferate readily on nanoporous glass membranes with mean pore sizes between 10 nm and 124 nm. In both proliferation and mRNA expression experiments, L929 fibroblasts show a distinct trend towards mean pore sizes > 80 nm. For primary hMSCs, excellent proliferation is observed on all nanoporous surfaces. hMSC on samples with 17 nm pore size display increased expression of COL10, COL2A1 and SOX9, especially during the first two weeks of culture. In upside down culture, SK MEL-28 cells on nanoporous glass resist the gravitational force and proliferate well in contrast to cells on flat references. The effect of paclitaxel treatment of MDA MB 321 breast cancer cells is already visible after 48 h on nanoporous membranes and strongly pronounced in comparison to reference samples. The studies presented in this work showed novel and distinct effects of micro- and nanoscale topographies on the behavior of various types of living cells. These examples display how versatile the potential for applications of bioactive materials could become in the next years and decades. And yet this variety of different alterations of cell functions due to topographic cues also shows the crucial part of this field of research: Carving out distinct, robust correlations of external cues and cell behavior is of utmost importance to derive definitive design implications that can lead to scientifically, clinically and commercially successful products.},
  language  = {en}
}
@article{BlancoKuchenbaeckerCuadrasetal.2015,
  author    = {Blanco, Ignacio and Kuchenbaecker, Karoline and Cuadras, Daniel and Wang, Xianshu and Barrowdale, Daniel and Ruiz de Garibay, Gorka and Librado, Pablo and Sanchez-Gracia, Alejandro and Rozas, Julio and Bonifaci, N{\´u}ria and McGuffog, Lesley and Pankratz, Vernon S. and Islam, Abul and Mateo, Francesca and Berenguer, Antoni and Petit, Anna and Catal{\`a}, Isabel and Brunet, Joan and Feliubadal{\´o}, Lidia and Tornero, Eva and Ben{\´i}tez, Javier and Osorio, Ana and Ram{\´o}n y Cajal, Teresa and Nevanlinna, Heli and Aittom{\"a}ki, Kristina and Arun, Banu K. and Toland, Amanda E. and Karlan, Beth Y. and Walsh, Christine and Lester, Jenny and Greene, Mark H. and Mai, Phuong L. and Nussbaum, Robert L. and Andrulis, Irene L. and Domchek, Susan M. and Nathanson, Katherine L. and Rebbeck, Timothy R. and Barkardottir, Rosa B. and Jakubowska, Anna and Lubinski, Jan and Durda, Katarzyna and Jaworska-Bieniek, Katarzyna and Claes, Kathleen and Van Maerken, Tom and D{\´i}ez, Orland and Hansen, Thomas V. and J{\o}nson, Lars and Gerdes, Anne-Marie and Ejlertsen, Bent and De la Hoya, Miguel and Cald{\´e}s, Trinidad and Dunning, Alison M. and Oliver, Clare and Fineberg, Elena and Cook, Margaret and Peock, Susan and McCann, Emma and Murray, Alex and Jacobs, Chris and Pichert, Gabriella and Lalloo, Fiona and Chu, Carol and Dorkins, Huw and Paterson, Joan and Ong, Kai-Ren and Teixeira, Manuel R. and Hogervorst, Frans B. L. and Van der Hout, Annemarie H. and Seynaeve, Caroline and Van der Luijt, Rob B. and Ligtenberg, Marjolijn J. L. and Devilee, Peter and Wijnen, Juul T. and Rookus, Matti A. and Meijers-Heijboer, Hanne E. J. and Blok, Marinus J. and Van den Ouweland, Ans M. W. and Aalfs, Cora M. and Rodriguez, Gustavo C. and Phillips, Kelly-Anne A. and Piedmonte, Marion and Nerenstone, Stacy R. and Bae-Jump, Victoria L. and O'Malley, David M. and Schmutzler, Rita K. and Wappenschmidt, Barbara and Rhiem, Kerstin and Engel, Christoph and Meindl, Alfons and Ditsch, Nina and Arnold, Norbert and Plendl, Hansjoerg J. and Niederacher, Dieter and Sutter, Christian and Wang-Gohrke, Shan and Steinemann, Doris and Preisler-Adams, Sabine and Kast, Karin and Varon-Mateeva, Raymonda and Gehrig, Andrea and Bojesen, Anders and Pedersen, Inge Sokilde and Sunde, Lone and Birk Jensen, Uffe and Thomassen, Mads and Kruse, Torben A. and Foretova, Lenka and Peterlongo, Paolo and Bernard, Loris and Peissel, Bernard and Scuvera, Giulietta and Manoukian, Siranoush and Radice, Paolo and Ottini, Laura and Montagna, Marco and Agata, Simona and Maugard, Christine and Simard, Jacques and Soucy, Penny and Berger, Andreas and Fink-Retter, Anneliese and Singer, Christian F. and Rappaport, Christine and Geschwantler-Kaulich, Daphne and Tea, Muy-Kheng and Pfeiler, Georg and John, Esther M. and Miron, Alex and Neuhausen, Susan L. and Terry, Mary Beth and Chung, Wendy K. and Daly, Mary B. and Goldgar, David E. and Janavicius, Ramunas and Dorfling, Cecilia M. and Van Rensburg, Elisabeth J. and Fostira, Florentia and Konstantopoulou, Irene and Garber, Judy and Godwin, Andrew K. and Olah, Edith and Narod, Steven A. and Rennert, Gad and Paluch, Shani Shimon and Laitman, Yael and Friedman, Eitan and Liljegren, Annelie and Rantala, Johanna and Stenmark-Askmalm, Marie and Loman, Niklas and Imyanitov, Evgeny N. and Hamann, Ute and Spurdle, Amanda B. and Healey, Sue and Weitzel, Jeffrey N. and Herzog, Josef and Margileth, David and Gorrini, Chiara and Esteller, Manel and G{\´o}mez, Antonio and Sayols, Sergi and Vidal, Enrique and Heyn, Holger and Stoppa-Lyonnet, Dominique and L{\´e}on{\´e}, Melanie and Barjhoux, Laure and Fassy-Colcombet, Marion and Pauw, Antoine de and Lasset, Christine and Fert Ferrer, Sandra and Castera, Laurent and Berthet, Pascaline and Cornelis, Fran{\c{c}}ois and Bignon, Yves-Jean and Damiola, Francesca and Mazoyer, Sylvie and Sinilnikova, Olga M. and Maxwell, Christopher A. and Vijai, Joseph and Robson, Mark and Kauff, Noah and Corines, Marina J. and Villano, Danylko and Cunningham, Julie and Lee, Adam and Lindor, Noralane and L{\´a}zaro, Conxi and Easton, Douglas F. and Offit, Kenneth and Chenevix-Trench, Georgia and Couch, Fergus J. and Antoniou, Antonis C. and Pujana, Miguel Angel},
  title     = {Assessing associations between the AURKA-HMMR-TPX2-TUBG1 functional module and breast cancer risk in BRCA1/2 mutation carriers},
  series = {PLoS ONE},
  volume    = {10},
  journal   = {PLoS ONE},
  number    = {4},
  doi       = {10.1371/journal.pone.0120020},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-143469},
  pages     = {e0120020},
  year      = {2015},
  abstract  = {While interplay between BRCA1 and AURKA-RHAMM-TPX2-TUBG1 regulates mammary epithelial polarization, common genetic variation in HMMR (gene product RHAMM) may be associated with risk of breast cancer in BRCA1 mutation carriers. Following on these observations, we further assessed the link between the AURKA-HMMR-TPX2-TUBG1 functional module and risk of breast cancer in BRCA1 or BRCA2 mutation carriers. Forty-one single nucleotide polymorphisms (SNPs) were genotyped in 15,252 BRCA1 and 8,211 BRCA2 mutation carriers and subsequently analyzed using a retrospective likelihood approach. The association of HMMR rs299290 with breast cancer risk in BRCA1 mutation carriers was confirmed: per-allele hazard ratio (HR) = 1.10, 95\% confidence interval (CI) 1.04 - 1.15, p = 1.9 x 10\(^{-4}\) (false discovery rate (FDR)-adjusted p = 0.043). Variation in CSTF1, located next to AURKA, was also found to be associated with breast cancer risk in BRCA2 mutation carriers: rs2426618 per-allele HR = 1.10, 95\% CI 1.03 - 1.16, p = 0.005 (FDR-adjusted p = 0.045). Assessment of pairwise interactions provided suggestions (FDR-adjusted p\(_{interaction}\) values > 0.05) for deviations from the multiplicative model for rs299290 and CSTF1 rs6064391, and rs299290 and TUBG1 rs11649877 in both BRCA1 and BRCA2 mutation carriers. Following these suggestions, the expression of HMMR and AURKA or TUBG1 in sporadic breast tumors was found to potentially interact, influencing patients' survival. Together, the results of this study support the hypothesis of a causative link between altered function of AURKA-HMMR-TPX2-TUBG1 and breast carcinogenesis in BRCA1/2 mutation carriers.},
  language  = {en}
}