Publications by authors named "Maria Duda"

7 Publications

  • Page 1 of 1

Genomics Integrated Systems Transgenesis (GENISYST) for gain-of-function disease modelling in Göttingen Minipigs.

J Pharmacol Toxicol Methods 2021 Feb 18;108:106956. Epub 2021 Feb 18.

Genome Biologics, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany; Institute of Cardiovascular Regeneration, Centre for Molecular Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany. Electronic address:

Göttingen Minipigs show several anatomical, physiological, and pathogenetical similarities to humans and serve an important role in translational studies for example as large animal models of disease. In recent years, the number of transgenic Göttingen Minipigs models has increased, as advanced genetic techniques simplify the generation of animals with precisely tailored modifications. These modifications are designed to replicate genetic alterations responsible for human disease. In addition to serving as valuable large animal disease models, transgenic Göttingen Minipigs are also considered promising donors for xenotransplantation. Current technologies for generation of transgenic minipigs demand a long development and production time of typically 2-3 years. To overcome this limitation and expand the use of Göttingen Minipigs for disease modelling and drug testing, we developed the GENISYST (Genomics Integrated Systems Transgenesis) technology platform for rapid and efficient generation of minipigs based transgenic disease models. As proof of concept, we report the successful generation of transgenic minipigs expressing green fluorescent protein (GFP) in multiple disease-relevant tissues including liver, heart, kidney, lungs, and the central nervous system (CNS). Our data demonstrates the feasibility, efficiency, and utility of GENISYST for rapid one-step generation of transgenic minipigs for human disease modelling in drug discovery and development.
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http://dx.doi.org/10.1016/j.vascn.2021.106956DOI Listing
February 2021

Planar Differential Growth Rates Initiate Precise Fold Positions in Complex Epithelia.

Dev Cell 2019 11 10;51(3):299-312.e4. Epub 2019 Oct 10.

MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK; Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK; College of Information and Control, Nanjing University of Information Science and Technology, Nanjing, Jiangsu 210044, China. Electronic address:

Tissue folding is a fundamental process that shapes epithelia into complex 3D organs. The initial positioning of folds is the foundation for the emergence of correct tissue morphology. Mechanisms forming individual folds have been studied, but the precise positioning of folds in complex, multi-folded epithelia is less well-understood. We present a computational model of morphogenesis, encompassing local differential growth and tissue mechanics, to investigate tissue fold positioning. We use the Drosophila wing disc as our model system and show that there is spatial-temporal heterogeneity in its planar growth rates. This differential growth, especially at the early stages of development, is the main driver for fold positioning. Increased apical layer stiffness and confinement by the basement membrane drive fold formation but influence positioning to a lesser degree. The model successfully predicts the in vivo morphology of overgrowth clones and wingless mutants via perturbations solely on planar differential growth in silico.
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http://dx.doi.org/10.1016/j.devcel.2019.09.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6838681PMC
November 2019

Stress relaxation in epithelial monolayers is controlled by the actomyosin cortex.

Nat Phys 2019 Aug 13;15(8):839-847. Epub 2019 May 13.

London Centre for Nanotechnology, University College London, Gower Street, London WC1E 6BT, UK.

Epithelial monolayers are one-cell thick tissue sheets that line most of the body surfaces, separating internal and external environments. As part of their function, they must withstand extrinsic mechanical stresses applied at high strain rates. However, little is known about how monolayers respond to mechanical deformations. Here, by subjecting suspended epithelial monolayers to stretch, we find that they dissipate stresses on a minute timescale and that relaxation can be described by a power law with an exponential cut-off at timescales larger than ~10 s. This process involves an increase in monolayer length, pointing to active remodelling of cellular biopolymers at the molecular scale during relaxation. Strikingly, monolayers consisting of tens of thousands of cells relax stress with similar dynamics to single rounded cells and both respond similarly to perturbations of the actomyosin cytoskeleton. By contrast, cell-cell junctional complexes and intermediate filaments do not relax tissue stress, but form stable connections between cells, allowing monolayers to behave rheologically as single cells. Taken together our data show that actomyosin dynamics governs the rheological properties of epithelial monolayers, dissipating applied stresses, and enabling changes in monolayer length.
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http://dx.doi.org/10.1038/s41567-019-0516-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116713PMC
August 2019

Polarization of Myosin II Refines Tissue Material Properties to Buffer Mechanical Stress.

Dev Cell 2019 01;48(2):245-260.e7

MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK; Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK; College of Information and Control, Nanjing University of Information Science and Technology, Nanjing, Jiangsu 210044, China. Electronic address:

As tissues develop, they are subjected to a variety of mechanical forces. Some of these forces are instrumental in the development of tissues, while others can result in tissue damage. Despite our extensive understanding of force-guided morphogenesis, we have only a limited understanding of how tissues prevent further morphogenesis once the shape is determined after development. Here, through the development of a tissue-stretching device, we uncover a mechanosensitive pathway that regulates tissue responses to mechanical stress through the polarization of actomyosin across the tissue. We show that stretch induces the formation of linear multicellular actomyosin cables, which depend on Diaphanous for their nucleation. These stiffen the epithelium, limiting further changes in shape, and prevent fractures from propagating across the tissue. Overall, this mechanism of force-induced changes in tissue mechanical properties provides a general model of force buffering that serves to preserve the shape of tissues under conditions of mechanical stress.
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http://dx.doi.org/10.1016/j.devcel.2018.12.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6353629PMC
January 2019

Tumor suppressor VHL functions in the control of mitotic fidelity.

Cancer Res 2014 May 20;74(9):2422-31. Epub 2013 Dec 20.

Authors' Affiliations: Institute of Molecular Health Sciences; Rodent Center HCI, Eidgenössische Technische Hochschule (ETH); and Institute of Surgical Pathology, University Hospital Zurich, Zurich, Switzerland.

The von Hippel-Lindau (VHL) tumor suppressor protein pVHL is commonly mutated in clear cell renal cell carcinoma (ccRCC) and has been implicated in the control of multiple cellular processes that might be linked to tumor suppression, including promoting proper spindle orientation and chromosomal stability. However, it is unclear whether pVHL exerts these mitotic regulatory functions in vivo as well. Here, we applied ischemic kidney injury to stimulate cell division in otherwise quiescent mouse adult kidneys. We show that in the short term (5.5 days after surgery), Vhl-deficient kidney cells demonstrate both spindle misorientation and aneuploidy. The spindle misorientation phenotype encompassed changes in directed cell division, which may manifest in the development of cystic lesions, whereas the aneuploidy phenotype involved the occurrence of lagging chromosomes but not chromosome bridges, indicative of mitotic checkpoint impairment. Intriguingly, in the long term (4 months after the ischemic insult), Vhl-deficient kidneys displayed a heterogeneous pattern of ccRCC precursor lesions, including cysts, clear cell-type cells, and dysplasia. Together, these data provide direct evidence for a key role of pVHL in mediating oriented cell division and faithful mitotic checkpoint function in the renal epithelium, emphasizing the importance of pVHL as a controller of mitotic fidelity in vivo.
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http://dx.doi.org/10.1158/0008-5472.CAN-13-2040DOI Listing
May 2014

Tumor suppressor NF2/Merlin is a microtubule stabilizer.

Cancer Res 2014 Jan 26;74(1):353-62. Epub 2013 Nov 26.

Authors' Affiliations: Institute of Molecular Health Sciences, ETH Zurich, Zurich, Switzerland; and Department of Cell Biology, The Scripps Research Institute, La Jolla, California.

Cancer-associated mutations in oncogene products and tumor suppressors contributing to tumor progression manifest themselves, at least in part, by deregulating microtubule-dependent cellular processes that play important roles in many cell biological pathways, including intracellular transport, cell architecture, and primary cilium and mitotic spindle organization. An essential characteristic of microtubules in the performance of these varied cell processes is their ability to continuously remodel, a phenomenon known as dynamic instability. It is therefore conceivable that part of the normal function of certain cancer-causing genes is to regulate microtubule dynamic instability. Here, we report the results of a high-resolution live-cell image-based RNA interference screen targeting a collection of 70 human tumor suppressor genes to uncover cancer genes affecting microtubule dynamic instability. Extraction and computational analysis of microtubule dynamics from EB3-GFP time-lapse image sequences identified the products of the tumor suppressor genes NF1 and NF2 as potent microtubule-stabilizing proteins. Further in-depth characterization of NF2 revealed that it binds to and stabilizes microtubules through attenuation of tubulin turnover by lowering both rates of microtubule polymerization and depolymerization as well as by reducing the frequency of microtubule catastrophes. The latter function appears to be mediated, in part, by inhibition of hydrolysis of tubulin-bound GTP on the growing microtubule plus end.
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http://dx.doi.org/10.1158/0008-5472.CAN-13-1334DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3929585PMC
January 2014

Systematic localization of the Arabidopsis core cell cycle proteins reveals novel cell division complexes.

Plant Physiol 2010 Feb 16;152(2):553-65. Epub 2009 Dec 16.

Department of Plant Systems Biology, Flanders Institute for Biotechnology, B-9052 Ghent, Belgium.

Cell division depends on the correct localization of the cyclin-dependent kinases that are regulated by phosphorylation, cyclin proteolysis, and protein-protein interactions. Although immunological assays can define cell cycle protein abundance and localization, they are not suitable for detecting the dynamic rearrangements of molecular components during cell division. Here, we applied an in vivo approach to trace the subcellular localization of 60 Arabidopsis (Arabidopsis thaliana) core cell cycle proteins fused to green fluorescent proteins during cell division in tobacco (Nicotiana tabacum) and Arabidopsis. Several cell cycle proteins showed a dynamic association with mitotic structures, such as condensed chromosomes and the preprophase band in both species, suggesting a strong conservation of targeting mechanisms. Furthermore, colocalized proteins were shown to bind in vivo, strengthening their localization-function connection. Thus, we identified unknown spatiotemporal territories where functional cell cycle protein interactions are most likely to occur.
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http://dx.doi.org/10.1104/pp.109.148643DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2815867PMC
February 2010