Pancreatic ductal adenocarcinoma (PDAC) is a deadly form of cancer that is on the rise due to various factors, including an aging population and unhealthy lifestyles. Regrettably, PDAC is among the top cancer killers, with a dismal five-year survival rate of less than 10%. The lack of adequate screening programs is a significant factor contributing to this dire statistic. However, the identification of PDAC in its early stages could help reduce the mortality rate by as much as 80%. The project LASERBLOOD (Biophotonic Nanoparticle-enabled Laser Blood Test for Early Detection of Pancreatic Cancer) aims to develop an in vitro diagnostic test based on the fluorescence lifetime fingerprint of the personalized protein corona, offering critical information at every stage of PDAC progression. The protein corona (PC) is a coating of bio-molecular substances surrounding nanoparticles when exposed to biofluids. It is both personalized and disease-specific, making it an ideal marker to monitor the variation of nanoparticle PC and correlate it to the development of PDAC. The analysis will use fluorescence lifetime (FL) analysis, a non-invasive, reactant-free, and real-time technique. In the initial phase, the consortium will utilize a mouse model (MKC) to identify the FL fingerprint of protein corona at each stage of PDAC development. The MKC mouse model is genetically engineered to be bioluminescent and develop PDAC in a controlled manner. By linking the development of PDAC observed through bioluminescent imaging to the FL response of PC in blood samples, the consortium will provide an unprecedented fingerprint of the disease's progression from its first occurrence. At a second stage, the project will validate on humans the use of the FL fingerprint of protein corona as a tool for the early diagnosis of PDAC. The findings will provide the scientific and technological foundation for the development of an in vitro PDAC test for large scale screening of the population.

Robust detection of single molecules in complex biological fluids is the ultimate goal in the field of disease biomarker analysis. Conventionally, to enable the quantitative analysis of individual molecules in macroscopic volumes, analyte pre-concentration and sample partitioning into fL-nL compartments has been combined with the amplification of the specific recognition events. In these setups, the positive or negative detection of fluorescence signal is triggered by enzymatic reactions occurring in each compartment. Binary readout based on Poisson statistics quantifies ultra-low concentrations of analyte molecules. This approach has been adopted for nucleic acids analysis in current digital PCR, and is also available for proteins in a technique coined as digital ELISA. The objective of VerSiLiB is to develop an enzyme-free amplification strategy for the analysis of both protein and nucleic acid analytes with the single digital platform that offers means to access additional information on target analytes not achievable with current technologies. Method is based on novel affinity-mediated-transport amplification, where affinity interaction of target analyte with a specific ligand attached to a magnetic nanoparticle transporter is accompanied with rapid shuttling of fluorescent tracers that serve as reporters. By applying external magnetic field, tracers are transported from the tracer storage side (where they are dark) to tracer active side (where they become bright) only if target analyte is present in the small reaction compartment. Tailored plasmonic nanostructures will be prepared at the storage and active sides of the compartment to render the tracer either dark or bright. The aim is to perform technology validation for the novel VerSiLiB proteogenomics amplification platform in cancer management using biobanked liquid biopsy samples.

Cancer biomarkers circulating in body fluids have been shown to reflect the pathological process and for this reason can be used for cancer diagnosis, prognosis and choice for therapeutic interventions. It is proven that their detection is a key to new minimal-invasive detection approaches. However, barriers to wide spread use of similar approaches are lack of test sensitivity, specificity and limited availability of low cost detection platforms. This project is focused at developing a compact plasmonic-based device with integrated microfluidic circuit and functionalized nanostructures for the detection of DNA, microRNA and tumor autoantibodies cancer biomarkers. The aim is to detect cancer biomarkers circulating in blood with improvement in sensitivity of factor up to 1000, reduction in cost of platform of factor ranging from 2 to 4 compared to today’s available techniques and analysis time less than 60 minutes. The proposed detection approache will provide ultrasensitive detection of biomolecular systems with no need for complex sample chemical modifications thus allowing direct and simple assays to be performed. Within the project a bimodal industrial prototype will be developed integrating novel surface plasmon resonance imaging and plasmon-enhanced fluorescence sensing technologies, respectively. Automated fabrication processes suitable for low cost mass production will be developed and applied to produce disposable integrated chips. Prototype will be specifically fabricated for early diagnosis and prognosis of colorectal cancer. The team includes partners holding cross-disciplinary competencies needed to achieve the proposed results, including two of the first five plasmon resonance groups in the world, the inventor of surface plasmon microscopy – also known as surface plasmon resonance imaging- and plasmon-enhanced fluorescence spectroscopy, and full European value chain including disposable chip and readout platforms design, development and manufacturing.

The mission of AiPBAND is to train a new generation of entrepreneurial and innovative early-stage researchers (ESRs) in the early diagnosis of brain tumours using molecular biomarkers in the blood, meeting the medical and societal challenges of this emerging field. AiPBAND will focus on gliomas, a range of devastating and progressive brain tumours affecting around 25,000 people each year in Europe and responsible for the majority of deaths from primary brain tumours. Fourteen fellows will be trained by experts in 9 academic and 3 non-academic beneficiaries, belonging to 5 EU member states and 6 partner organizations (4 private sectors and 2 international academic), with fields ranging from neuroscience, engineering (including big data science), healthcare to economics. State-of-the-art technologies will be applied in parallel to (i) identify novel blood biomarkers from patients with gliomas, (ii) design three types of multiplex biosensor (plasmonic-based, graphene-based, and digital ELISA assay-based), (iii) develop a big data-empowered intelligent data management infrastructure, and (iv) develop cloud-based diagnostic systems. Proof-of-concept will be evaluated through clinical trials to assess accuracy, sensitivity and specificity. The elaborately designed individual research projects under the Vitae Researcher Development Framework – carefully arranged into local training courses, network wide events, secondments, personalized career development plans, with strong involvement of the private sector – will ensure exploitation of AiPBAND's achievements, and will maximize the ESRs’ abilities in creative & innovative thinking, triple-i knowledge transformation, and encourage a business-orientated mind-set and entrepreneurship.