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Biomechanics in health and disease: advanced physical tools for innovative early diagnosis

Periodic Reporting for period 2 - Phys2BioMed (Biomechanics in health and disease: advanced physical tools for innovative early diagnosis)

Reporting period: 2021-01-01 to 2022-12-31

The identification and understanding of the physical and chemical determinants of diseases, typically at the level of tissues and their constituents (cells and extracellular matrix, ECM), represent a potential breakthrough in nanomedicine because it would allow developing effective early diagnostic methods and, ideally, preventive strategies. In particular, there is a growing evidence of the role of mechanics in cell biology related to various stresses and diseases. Phys2BioMed aims therefore at identifying and collecting mechanical markers, which allow for the early detection of cell and tissue dysfunction and the onset of diseases.

Phys2BioMed will offer excellent interdisciplinary and cross-sectoral training to a team of motivated early stage researchers (ESRs) on the application of cutting-edge physical tools for the mechanical phenotyping of cells and tissues of clinical relevance, aiming at developing novel early-diagnostic tools.
A key element is the peer-to-peer collaboration of research academic institutions with industries and world-leading medical and clinical centers, who will highlight unmet clinical needs.

Phys2BioMed will provide scientific and technological outcomes on biomechanics, and the mechanical determinants of diseases, by:
- Defining standardized procedures for nanomechanical measurements, and the main features of new-generation instrumentation optimized for the mechanical phenotyping of clinical specimens;
- providing, in the longer-term, the platform and know-how to build a data bank of mechanical fingerprints of diseases, for the development of effective early-diagnostic tools.

Specific scientific and technological objectives of the research programme:
- To establish the mechanical and rheological fingerprints of cells and tissues and their alterations in diseases. Phys2BioMed will produce comprehensive nano-mechanical and rheological phenotypes of cells and tissues to define mechanical fingerprints of specific diseases. The main approach used will be based on atomic force microscopy (AFM) coupled to different methods such as microfluidic cytometry, non-AFM nano-indentation and structural inspection.
- To establish procedures, models, and standards for assessments of cell and tissue nano-mechanical properties. The approach will reside in defining standard procedures for mechanical tests, sample preparation and data analysis. Databases will be launched to be open to the scientific community.
- To correlate the output of the mechanical and rheological phenotyping of clinical samples (ECMs, cells, tissues) to specific clinical features of the selected diseases, so to define heuristic mechanical fingerprints to be used as early-diagnostic cues.
After the initial effort for selecting and recruiting the 15 Early Stage Researchers (ESRs) during the first 10 months of the project, the scientific activity has started at the different nodes of the network and is gaining momentum.
On the fundamental-science level, the partners have identified the links between biomechanics and disease. This work was based on a comprehensive analysis of the current literature, and is meant to serve as a reference knowledge toolkit for the ESRs. The aim of this work is to identify general links between cell and tissue biomechanics and different diseases, and to provide characterization of the differential mechanical features in healthy and diseased cells and tissues. The knowledge gained and rationalised during this effort was compiled in a book about "Mechanics in Disease", which will be published by De Gruyter in two volumes(Methods and Applications) in February 2023.
The partners have also defined standardised sample preparation and shipping procedures. In particular, this effort was devoted to establish a common ground of best practices for preparing cellular specimens (tissues will be considered later in the project) that are suitable for the mechanical analysis, but also easily transferable among different laboratories and between the clinics and the labs.
Also, standardised procedures for cell mechanics have been discussed and defined, with emphasis on reliable and common practices for calibration of atomic force microscopy probes.
The standardisation of mechanical measurements on cells have been completed. Cellular samples have been prepared, shipped and measured according to the established procedures. Data analysis has been done. A similar standardisation effort on clinically-relevant murine bladder tissues has been completed. Two publications are in advanced preparation stage.
Another important task initiated was the definition and implementation of a universal file format (UFF) to share the force curves obtained during the research developed within this project. Moreover, a toolkit of Python scripts for performing mechanical data analysis has been developed.
Besides scientific research, the network takes advantage of a fully functional website, cloud and Wiki, where discussion forums support and animate the internal discussion.
A MOOC (massive online open course) has been prepared.
The partners have organized a final Symposium as conclusive event of Phys2BioMed. During the Symposium "Present and future of mechanics of cells and tissues in health and disease", ESRs and invited speakers will present their contributions under three main topics: methods for cell and tissue mechanics: AFM and beyond; the mechanical phenotype of cells in health and disease; cell and ECM: a complex mechanical interplay.
Cell and tissue mechanics used for detecting diseases is a new paradigm, which requires the standardisation of novel and robust scientific and methodological approaches in order to be effective.
This is a prerequisite to be eventually accepted in the medical/clinical environment and positively impacting on the society. Phys2BioMed aims at narrowing the distance towards this ambitious objective.
Bringing cell mechanics towards the the clinics is still a challenge. The first steps of using the mechanical phenotyping to obtain diagnostic, or prognostic markers, are first the standardisation of the approaches at the multi-laboratory level, and then the validation of the applicability of the standardised approach to clinical, samples, (the standardisation of the protocol at the clinical level). This objective represents a relevant progress beyond the state of the art.
The impact of Phys2BioMed activity is expected to be high by virtue of the composition of the network: the academic partners will be supported by strongly research and technology–oriented private companies who are leaders in the development of dedicated instrumentation for (bio)mechanical measurements, as well as in the transfer of state-of-the-art technologies outside the academy into the private sector. Moreover, specific methodologies and skills of the health public and private system are brought into the project by the medical/clinical beneficiaries and partner organisations. Phys2BioMed creates the common place where confrontation between such diverse players can take place.
Phys2BioMed will enhance the intersectoral and public-private collaborations in the healthcare field, in particular, between the academia and medical/clinical and industrial subjects. This will favour, in the medium term, the development of new approaches to fight diseases, and new products and instrumentation for screening, diagnosis and therapy.
ESRs and Beneficiaries at the 7th Phys2BioMed Progress Meeting in Gargnano, Italy, May 2022
Topographic and mechanical mapping of cells
ESRs and some beneficiaries during a social event of the 7th PM in Gargnano, Italy, May 2022
Nanoindentation
AFM-based nano mechanics
ESRs and Beneficiaries at the Final Meeting in Barcelona, Spain, Nov. 2022
ESRs and Beneficiaries at the 1st Progress Meeting and Int. Conf. (AFMBIOMED) in Munster, Sept. 2019
Phys2BioMed logo
ESRs and Beneficiaries at the 1st Phys2BioMed school in Grenoble, October 2019