TOPOGRAPHYSENSING · Effects of 3D topographies on mechanosensing in intestine epithelial architecture and dynamics
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2019-09-01 → 2021-08-31
- EU contribution
- €134,600
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
Lines connect the coordinator with its partners.
Results in brief
Effects of 3D topographies on mechanosensing in intestine epithelial architecture and dynamics
The intestine epithelium (IE) consists of spatially segregated cells that organize into groups of various functions at different locations of the 3-dimensional (3D) curved epithelial monolayer. How geometric cues contribute to the maintenance of the sophisticated epithelial architecture and dynamics in 3D remains unknown until now. Thus, a systematic investigation on the effect of curvature on IE architecture, organization, and dynamics in the context of 3D mechanosensing is highly needed to significantly improve our understanding of normal IE structure maintenance and homeostasis, etc. In this project, I developed a novel 3D intestinal model based on biomimetic hydrogels that allow the long-term culture of primary intestinal cells. Substrate elasticity was designed to match the mechanical properties of the physiological extracellular matrix (ECM). By applying two-step lithography and soft molding methods, I produced 3D scaffolds reflecting in vivo intestinal 3D architectures. I then grew primary intestinal stem cells on these scaffolds to realize in vivo-like compartmentalization and tissue structure. With this, I systematically studied 3D cellular dynamics and cytoskeleton organization on out-of-plane curvature. Furthermore, I could conveniently integrate the cell-laden scaffolds into a two-layer microfluidic system, which offers a practical approach for monitoring epithelial remodeling processes in 3D contexts. This new platform can possibly be translated into a disease model for studying some rare intestinal diseases, such as Congenital Tufting Enteropathy (CTE). The outcome of this action then has the potential to influence fields in fundamental cell development research and therapeutic studies. It could also become a successful paradigm for other tissue engineering studies.
Data: CORDIS, © European Union
Project objective
Intestine epithelium consists of spatially segregated cells that organize into groups of various functions at different locationsof the 3D curved epithelial monolayer. How geometric cues contribute to the maintenance of the sophisticated epithelial architecture and dynamics in 3D remains unknown until now. Recently, the Ladoux's laboratory has found that EpCAM-modulated cell contractility associated with the epithelial monolayer polarity, cytoskeletal arrangement, and cell-cell adhesion in 3D context. In contrast to 2D context, the EpCAM-defective tissue shows a loss of collective cellular spatial organization and forms a disordered multi-layered epithelium when exposed to substrates of 3D topographies. In addition, Ankyrin-G and α/β-spectrin network which participates in cortical tension modulation was identified as the main interacting partner with EpCAM in epithelial cells. These observations lead us to hypothesize that EpCAM allows the tissue to sense and conform to complex 3D topographies in an orderly manner. However, the molecular mechanisms and other related functions of EpCAM-mediated mechanotransduction remain unknown. As large scale mechanosensing has been shown to occur primarily through the actin cytoskeleton which permeates the tissue to form a network, we aim to understand the interactions between the EpCAM-mediated pathway and actin modulation and/or E-cadherin adhesion sites that may allow 3D topographical sensing. Our working hypothesis is that EpCAM forms an integral part of the cellular responses to topographic cues that has a more general role in controlling epithelial architecture and dynamics through the regulation of actomyosin networks, or vice versa. Here, we propose to scrutinize EpCAM-mediated mechanotransduction by generating a platform with precise control of geometric factors and microenvironmental cues using a range of multidisciplinary approaches including microfabrication, biophysics, and advanced molecular biology techniques.
Original text from CORDIS.
Participants
- CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS · ParisCoordinatorFrance
Links
Data: CORDIS, © European Union