Does inter-tissue mechanical coupling coordinate neural tube closure?

Project Information

NeuroForceSensor

Grant agreement ID: 101067028

DOI

10.3030/101067028

Project closed

EC signature date 28 July 2022

Start date 1 November 2022

End date 31 October 2024

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 179 438,08

Coordinated by

UNIVERSITA DEGLI STUDI DI PADOVA
Italy

Periodic Reporting for period 1 - NeuroForceSensor (Does inter-tissue mechanical coupling coordinate neural tube closure?)

Reporting period: 2022-11-01 to 2024-10-31

Summary of the context and overall objectives of the project

Neural tube defects (NTDs) are severe congenital malformations of the brain and spine, affecting 1 in 1000 births in Europe. They result from failure of the embryonic neural tube to close, leading to lifelong disabilities. While the cellular and molecular mechanisms are well-studied, the mechanical aspects of neural tube closure are less understood. The project aims to provide a unified biomechanical understanding of neural tube morphogenesis using advanced bioengineering techniques such as intravital 3D bioprinting. This technique enables the construction of force sensors directly inside the neural tube of chicken embryos, which bear striking similarities to the human embryo in early development. The first objective is to map tissue-level mechanical forces during neural tube closure. This will provide an overview of how forces and mechanical properties change over time and between different anatomical locations in the embryo. The second objective is to investigate whether connected epithelia are mechanically coupled, using precise physical perturbations and microfluidic delivery of pharmacological inhibitors. Together, these studies will pave the way for the identification of novel preventive and therapeutic strategies to enhance cellular force-generating mechanisms during neural tube closure. In parallel to data acquisition, the MSCA fellowship aims to foster the development of the individual researcher and provide interdisciplinary training and transferable skills.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The research project was conducted in the BioERA lab, under the supervision of Prof Elvassore. The lab is integrated between the Department of Industrial Engineering (UniPD) and the Veneto Insitute of Molecular Medicine (VIMM) in Padova. Work performed during the project was focused on the development and validation of a method for direct and dynamic quantification of mechanical forces in the chick embryo. This involved i3D bioprinting and finite element modelling (FEM) to define the structural and material properties of the force sensors and obtain measurements with accuracy in the nano-Newton range. The results led to a high impact first author publication (Maniou et al., 2024 Nature Materials). Adaptation of the sensors along the embryonic axis revealed that, unlike the cranial region where neural fold narrowing is predominant, closure in the spinal neural tube is governed by flow of the neuroepithelium. The next aim was to assess biomechanical coupling between the neuroepithelium and surface ectoderm (future skin). For this, I developed an application of hydrogel microfluidics, enabling targeted delivery of pharmacological compounds to the tissue of interest (manuscript in preparation 2025). The non-scientific deliverables of the project were fully achieved including training, transfer of knowledge, dissemination, communication, and project management. Results generated during this MSCA were reported in: (1) a first author publication (mentioned above) and a pre-print (2) selected presentations at five international conferences (3) more than ten articles in news and blogs (4) four communication activities including a research open day and an article on the future of developmental biology. International collaborations established or maintained during the project will ensure the continuity of knowledge exchange and support future joint initiatives.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

- This MSCA has pushed the frontiers of mechanobiology and constituted a step stage in our understanding of neural tube morphogenesis. As acknowledged by the scientific community, it has provided the clearest profiling of the forces that promote or counteract neural tube closure to date. Dissemination efforts have elicited strong interest, particularly in stem cell research and regenerative medicine.
- Mapping mechanical tensions along the embryonic axis, uncovered mechanistic differences between cranial and spinal neural tube closure. This could explain why NTD-predisposed embryos may develop exencephaly or spina bifida in isolation. Incorporation of the data collected into finite element models is helping to associate dynamic changes in mechanical properties and tissue displacement in early embryo development.
- The project introduced Developmental Biology and Biomechanics to the University of Padova through the establishment of an embryology lab, student training and supervision. It also combined expertise in chemical and tissue engineering and computational modelling, promoting multidisciplinary research.
- Knowledge generated will contribute to the organoid field, informing the generation of more reproducible organoid shape and patterning. The findings will be associated with neuronal differentiation and disease modelling applications.
- International mobility during the Action helped in establishing a strong network of collaborators, raising the profile of the host lab and the University.

Cylindrical force sensor, 3D printed into the developing spinal cord of a chicken embryo.

Elvassore lab retreat 2024, Colli Euganei, Italy