Sedzinski Group - Mechanics of Tissue Homeostasis

The Sedzinski group aims to understand the general principles underlying epithelial homeostasis. Particularly, they want to determine both the mechanics and molecular regulation of epithelial cell renewal from stem cells hoping to provide new insights into treatment of epithelial pathologies associated with defective tissue homeostasis, such as asthma, colitis, and most common epithelial tumours, or carcinomas. 

 

 

Apical emergence – a new cell behavior

During epithelial cell renewal, a nascent cell must not only define an apical domain, but must also build a new apical surface of sufficient size to accommodate its function (e.g. directional beating in a ciliated cell, luminal secretion in a secretory cell, etc.). This process of apical surface emergence can be envisioned perhaps most simply as the converse of apical constriction, a well-defined cell behavior underlying epithelial cell extrusion and epithelial folding. While molecular and mechanical aspects of apical constriction have been extensively studied, far less is known of apical emergence.

One major unresolved question concerns force generation during apical emergence of individual cells. What are the mechanisms that expand the growing apical surface? Moreover, how do new cells generate force to displace the neighboring cells that abut them? These questions about addition of new cells to epithelia are important because the answer will provide an essential complement to the burst of recent studies elucidating mechanisms of force generation during extrusion of old cells from epithelia and most importantly will provide key insights into mechanisms of epithelial homeostasis. 

Figure 1. Apical emergence is an essential step for epithelial cell renewal.

A) Schematics of epithelial cell renewal. Basally localized progenitors move (step 1 to 3) from basal to apical layer of a multilayered epithelium. Upon docking at the tricellular junction, cell progenitors integrate within an existing epithelium by expanding their apical domain (step 3 to 4). B) We use Xenopus laevis mucociliary epithelium as a model system to study epithelial cell renewal in vivo (picture courtesy of Wallingford lab). C) Schematics of apical emergence. Green – cell progenitor, grey  - existing epithelium. Apical domain of the progenitor cell expands displacing the neighboring cells. D, E) Image sequence of apically emerging multiciliated cell (MCC) (visualized with LifeAct-GFP, green) within Xenopus laevis epidermis (visualized with LifeAct-RFP). Note that cell-type specific promoters allow for targeted protein expression to MCCs of surrounding cells only.

Theoretical model of apical emergence

In collaboration with Dr. Edouard Hannezo we develop a minimal theoretical model of apically emerging cell. The model says that actin-based pushing (effective 2D Pressure), pulling (Λ), and contractile (𝛾) forces control expansion of the apical domain. The respective contributions of these forces can be assessed by analyzing the shape and dynamics behavior of the apical cell perimeter.

Figure 2. Theoretical model of apical emergence.

A) Apical domain expanding forces: effective 2D Pressure (2D Pressure) and neighboring junctional pulling forces (Λ) acting against cortical tension (𝛾) and elasticity from the surrounding cells (E). B) Model predicts shape changes of the apical domain and expansion dynamics. When considering cell autonomous forces within the cell apex, apical surface perimeters are rounded when pushing forces dominate, but are polygonal when pulling forces are in excess. Moreover, apical perimeters collapse when cortical tension (𝛾) exceeds the pushing forces (2D). For intermediary pressure, third cell behavior appears, where the apical area overshoots its steady state value, then undergoes damped oscillations.

Mechanism of apical actin assembly

Mature multiciliated cells are characterized by a complex and well-defined apical actin network. This stereotypic pattern is generated by the action of actin-binding proteins, such as actin nucleators and cross-linkers. The coordinated action of these proteins could generate the pushing force that expands apical surface. We use TIRF and super-resolution microscopy to image the process of apical actin assembly.

Figure 3. Assembly of the apical actin network expands the apical domain of a progenitor cell. A) Schematics of the apical actin network assembly. B) Imaging of de novo actin network assembly in vivo.

 

Tu, F., Sedzinski, J., Ma, Y., Marcotte, E.M., and Wallingford, J.B. (2018). Protein localization screening in vivo reveals novel regulators of multiciliated cell development and function. Journal of Cell Science 131, doi:10.1242/jcs.206565.
The cover shows a single multiciliated cell (MCC) within Xenopus laevis (frog) embryonic epidermis. This system allows understanding the function of unknown or poorly-studied proteins by determining their localization within a cell. In this study, 360 unknown genes were fluorescently-tagged and expressed within MCCs. These proteins localize to the specific domains within a cell, indicating the function of a gene. Click on the image to enlarge

Sedzinski, J., Hannezo, E., Tu, F., Biro, M., and Wallingford, J.B. (2017). RhoA regulates actin network dynamics during apical surface emergence in multiciliated epithelial cells. Journal of Cell Science 130, 420-428, doi:10.1242/jcs.194704. Correction in Journal of Cell Science 130, 1017. doi: 10.1242/jcs.202234.

Sedzinski, J., Hannezo, E., Tu, F., Biro, M., and Wallingford, J.B. (2016). Emergence of an Apical Epithelial Cell Surface In Vivo. Developmental Cell 36, 24-35, doi:10.1016/j.devcel.2015.12.013.

Sedzinski, J., and Wallingford, J. (2015). Planar Pol(o)arity. Developmental Cell 33, 494-495, doi:10.1016/j.devcel.2015.05.022.

Turk, E., Wills, A.A., Kwon, T., Sedzinski, J., Wallingford, J.B., and Stearns, T. (2015). Zeta-Tubulin Is a Member of a Conserved Tubulin Module and Is a Component of the Centriolar Basal Foot in Multiciliated Cells. Current Biology 25, 2177-2183, doi:10.1016/j.cub.2015.06.063.

Sedzinski, J., Biro, M., Oswald, A., Tinevez, J.Y., Salbreux, G., and Paluch, E. (2011). Polar actomyosin contractility destabilizes the position of the cytokinetic furrow. Nature 476, 462-466, doi:10.1038/nature10286.

 

Research Profile

Jakub Sedzinski’s education and training have been cross-disciplinary, spanning topics in biology, biophysics, and computational biology. After undergraduate studies in Molecular Biology at the University of Wroclaw in Poland, he completed a PhD at the Max Planck Institute of Molecular Cell Biology and Genetics in Dresden with Professor Ewa Paluch. There he became fascinated with biophysical approaches to cell biology and investigated the basics mechanisms of cytokinesis. He discovered that cytokinesis is inherently unstable and that imbalance in contractile forces compromises cytokinetic furrow positioning.

He then received a prestigious fellowship from European Molecular Biology Organization (EMBO) to carry out research at The University of Texas at Austin in the laboratory of Prof. John Wallingford. Here, Jakub continued his training in mechanics and regulation of cell shape, but in the context of developing embryos. His postdoctoral work defined a novel cell behavior - apical emergence, which describes the broad spectrum of cellular processes resulting in integration of a progenitor cell with an existing epithelium.

The Sedzinski laboratory at DanStem is now focused on understanding the molecular and mechanical mechanisms of epithelial cell renewal in vivo.

About Jakub Sedzinski

Curriculum Vitae

January 2022: Associate Professor, Group Leader
University of Copenhagen, Denmark
Novo Nordisk Foundation Center for Stem Cell Medicine, reNEW

March 2017: Associate Professor, Group Leader
University of Copenhagen, Denmark
Novo Nordisk Foundation Center for Stem Cell Biology, DanStem

June 2016 - March 2017: Research Associate
The University of Texas at Austin, USA
Advisor: Prof. John Wallingford

September 2011 - June 2016: Post-Doctoral Training
The University of Texas at Austin, USA
Advisor: Prof. John Wallingford

December 2010 - August 2011: Post-Doctoral Training
The Max Planck Institute of Molecular Cell Biology and Genetic, Dresden, Germany
Advisor: Prof. Ewa Paluch

September 2006 - December 2010: Doctorate in Biology
The Max Planck Institute of Molecular Cell Biology and Genetic, Dresden, Germany
Advisor: Prof. Ewa Paluch

July 2005 - June 2006: MSc in Biology
The University of Wroclaw, Poland
Advisor: Prof. Jan Szopa

September 2004 - June 2005: Visiting scientist
The Max Planck Institute of Molecular Plant Physiology, Golm, Germany
Advisor: Dr. Rita Zrenner

September 2001 - July 2004: BSc in Biology
July 2004 The University of Wroclaw, Poland
Advisor: Prof. Jan Szopa

 

 

Jakub Sedzinski: Mucociliary epithelium - the airways' wash station

 

Staff List - Sedzinski Group

Name Title Job responsibilities Phone E-mail
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Bussek-Sedzinski, Alexandra Maria Academic Research Staff Academic Employee   E-mail
Pasias, Athanasios Research Assistant   E-mail
Proks, Martin Research Assistant   E-mail
Sedzinski, Jakub Group Leader Associate Professor, Group Leader +4593565782 E-mail
Sørensen, Madison Lauren External   E-mail
Thiagarajan, Raghavan Assistant Professor Postdoc   E-mail
Thurner, Larissa Alina PhD Fellow +4535321641 E-mail
Tolonen, Mari Johanna Katariina PhD Student +4535336191 E-mail
Ventura, Guilherme Bastos Research Assistant Research Assistant   E-mail