Central to this thesis was the use of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to model a rare form of mitochondrial cardiomyopathy. To model this disease required the... Show moreCentral to this thesis was the use of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to model a rare form of mitochondrial cardiomyopathy. To model this disease required the design of new methodologies, improving upon the current limitations of hiPSC-CMs as model systems, notably the variability and the immature state of the resulting cardiomyocytes. The diseases we were interested in manifest soon after birth. At the outset of the studies, it was unclear whether a phenotype would be evident in our standard immature 2D cultures, or whether more complex models would be required to capture more salient features of the condition. Our focus was on the rare mitochondrial disease Combined Oxidative Phosphorylation Deficiency, caused by mutations in the gene alanyl-tRNA synthetase 2. We refined our emerging 3D cardiac microtissue protocol to improve robustness and reproducibility and reduce cost by basing differentiation on small molecules rather than growth factors without altering the functionality of hiPSC-CMs. We also demonstrated that integrating pH and O2 sensors in a micro-physiological chip was possible for the assessment of metabolic parameters under microfluidic flow. The overall goal of this thesis was to provide additional tools that would have utility in studying mitochondrial and other cardiac diseases. Show less
Jong, M.A. de; Adegeest, E.; Bérenger-Currias, N.M.L.P.; Mircea, M; Merks, R.M.H.; Semrau, S. 2023
The work included in this thesis is aimed at developing novel tools to advance our understanding of prostate cancer. The clinical problem of prostate cancer is presented and discussed in the wider... Show moreThe work included in this thesis is aimed at developing novel tools to advance our understanding of prostate cancer. The clinical problem of prostate cancer is presented and discussed in the wider context of the current clinical knowledge, highlighting the genetic mechanisms at its base. A dedicated chapter focuses on bone metastases, highly morbid feature of advanced prostate cancer, discussing the known mechanisms and the available models to study it in translational research. Then, moving from the molecular analysis of clinical specimens of bone metastasis, a biochemical pathway is identified and further studied in vitro, ex vivo and in vivo, validating the initial findings. A novel, early-stage prostate cancer patient-derived xenograft is presented and extensively characterized and implemented in a drug screening. This allowed to screen the effect of over 70 known drugs on prostate cancer models, using three-dimensional cultures and a semi-automated platform. As all research builds on previously established findings, a bibliometric analysis tool is presented, to assist in the generation of a knowledge network arranged by topic and impact of research. All these aspects and findings are then discussed in the context of the current direction of prostate cancer research, its emerging tools and its long-known challenges. Show less
Balak, J.R.A.; Juksar, J.; Carlotti, F.; Nigro, A. lo; Koning, E.J.P. de 2019
Purpose of Review Novel 3D organoid culture techniques have enabled long-term expansion of pancreatic tissue. This review comprehensively summarizes and evaluates the applications of primary tissue... Show morePurpose of Review Novel 3D organoid culture techniques have enabled long-term expansion of pancreatic tissue. This review comprehensively summarizes and evaluates the applications of primary tissue-derived pancreatic organoids in regenerative studies, disease modelling, and personalized medicine.Recent Findings Organoids derived from human fetal and adult pancreatic tissue have been used to study pancreas development and repair. Generated adult human pancreatic organoids harbor the capacity for clonal expansion and endocrine cell formation. In addition, organoids have been generated from human pancreatic ductal adenocarcinoma in order to study tumor behavior and assess drug responses.Summary Pancreatic organoids constitute an important translational bridge between in vitro and in vivo models, enhancing our understanding of pancreatic cell biology. Current applications for pancreatic organoid technology include studies on tissue regeneration, disease modelling, and drug screening. Show less
