The aim of ‘AMNIOGEL’ is the development of human extracellular matrix (ECM) based materials, as radically innovative, highly versatile human-derived platforms for 3D cell culture, microtissue development and disease models establishment. In particular, we will develop 3D disease models that could be used as an enabling tool for personalized drug discovery, increasing our understanding of the mechanisms behind bone cancer. The proposed cutting-edge technology enables the culture of human cells in a physiologically relevant microenvironment for applications in cell culture research, drug screening and development, cancer research, tissue engineering, replacement of animal testing and therapeutic applications. The potential of this technology to reduce or completely replace use of animals for biological screenings is expected to have a significant impact in the 3D cell culture market and pharmaceutical industry by accelerating drug screening and development reducing associated costs. The innovation potential of our products is based on the fact that contain human biochemical cues (vital for cell function), is a complete xeno-free solution (avoids contamination) for human cell culture and easy to manipulate. It is worth mentioning that offers the possibility to be personalised (using the patient’s own ECM) according to the customer needs. Moreover, our culture substrates are easily processed in multiple geometries and in microarrays amenable to ‘organ-on- a-chip’ systems designed for high-throughput screening (HTS) applications.

The last decades witnessed a quest for devices responding to temperature at a distance with unprecedented space resolution, approaching the nanoscale. Such devices are valuable in both fundamental and applied science, from overheat in micromachines to hyperthermia applied to cells. Despite great advances, the response is still collected in 2D. In real systems, heat flows in 3 dimensions such that 2D nanothermometers give just a plane view of a 3D reality. The restriction to 2D emerges because space resolution is bound to time and temperature resolutions, leading to a trilemma: scanning into the 3rd dimension is time consuming and cannot be achieve without losing temperature and time resolutions. While incremental improvements have been achieved in recent years, adding the 3rd dimension to nanothermometry is crucial for further impact and requires an innovative approach. Herein, I propose the development of nano local probes with tailored magnetic properties recording critical information about local temperature in 3D. These thermometric local probes avoid the resolution trilemma by recording the most relevant temperature information instead of reading the present temperature value. In many applications, including cellular hyperthermia, most part of the current temperature reading is of minor relevance and can be dropped. The key temperature information includes the maximum temperature achieved, the surpass of a given temperature threshold, and the time elapsed after this surpass. Once recorded, this key information can be read in 3D by standard devices (such as confocal microscopes and magnetic resonance imaging scanners) without time constrains and thus keeping a high space and temperature resolution. Moreover, the reading step can be performed in-situ and/or ex-situ, decoupling probes and reading devices if needed. This widens the range of applications of nanothermometers, allowing detection in confined environments and in non-transparent media.

This project addresses the quest of new materials and approaches that nanotechnology requires to solve the current limitations of medicine. The potential to externally track and image organs and the potential presence and evolution of diseases by using light becomes a reality thanks to the use of especially tailored biocompatible nanoplatforms. New avenues to image living bodies by light allied with mechanical waves (i.e. sound) are going to be opened. We propose here an elegant marriage between light and sound endowing smartly designed nanoprobes with the capability of deep-tissue photoacoustic imaging, also accompanied by all-optical temperature sub-tissue measuring. To increase the penetration depth and spatial resolution, an imaging approach pumping the probes with near-infrared light is proposed. Therefore, we propose a kind of material no previously exploited for photoacoustic exogenous agent, working as bimodal nanoprobe by also optically measuring temperature within the biological transparency windows in the near-infrared

Ship Clones: Characterization and search for biomarker of marine transmissible cancers Clonally transmissible cancers are cell clones that are transmitted between individuals via the transfer of living cancer cells. There are only three known types of naturally occurring clonally transmissible cancers, one of which is a disseminated neoplasia (DN) found in marine bivalves. DN is a leukemia-like cancer characterized by the proliferation of hypertrophied cells. In later stages of the disease, these cells spread throughout all tissues destroying their normal architecture and causing the death of their host organism. A polyphyletic origin of the DN has been detected in the common cockle Cerastoderma edule. DN has appeared on at least six different occasions and the six clones currently known coexist in cockle populations of the European Atlantic coast. In the present proposal high resolution lipidomics and proteomics will be used to screen for clonal biomarkers of DN and help to better understand the biological systems of transmissible cancers. Marine transmissible cancers are interesting biological models to better understand cancer transmissibility and metastasis. Marine clones likely ship using ocean currents to colonize new hosts in different regions. It is likely that they may also be unintentionally introduced by the action of man in disease-free regions. Hence, these cancers represent a potential threat for marine ecosystems, as they negatively impact keystone species (bivalves), that also have important socio-economic value for fisheries. As such, it is paramount to successfully identify, monitor and characterize the prevalence of clonal DN in marine bivalves, before any successful prevention and mitigation actions can be applied.

Bioprinting techniques, which integrate 3D printing with tissue engineering by using living cells encapsulated in natural or synthetic biomaterials as bioinks, are paving the way toward devising many innovating solutions for key biomedical and healthcare challenges and heralds' new frontiers in medicine, pharmaceutical, and food industries. HumanINK aims to validate human based-bioinks to produce robust humanized 3D environments with unprecedented biofunctionality for cell culture that fully recapitulate the native microenvironment of a variety of human tissues and organs. Under this project human-protein derivative precursors that can be cured upon light exposure to form soft hydrogels with tunable mechanical properties will be tested and validated as bioinks for 3D bioprinting. Such materials provide functional support for cell growth and interact with cells to control their function, guiding the process of tissue morphogenesis. This platform is the first to offer complete human-based material for bioprinting and an easy-to-use solution to create physiologically relevant 3D in vitro cell cultures, accelerate drug discovery or clinical purposes. The HumanINK will allow to optimize the printability, robustness, reproducibility and scalability of the human-based bioinks. The biological response of multiple human cell types will be investigated and the bioinks will be benchmarked with the main competitors in the market. Our proposed technology will increase the probability of successful drug development while simultaneously reducing the cost and time of development and supporting animal welfare, reducing animal experimentation. Based on the unique properties of our products, HumanINK represents a unique opportunity to develop materials for tissue engineering and accurate disease models for bridging the gap between fundamental research and drug validation, with a high and broad market potential in pharma companies, clinical institutions, or research groups.