Research Project

RESEARCH PROJECT

The Consorzio Interuniversitario in Ingegneria e Medicina (COIIM), founded in 2019, The consortium actively supports the research projects of the University of Molise, in particular the Department of Medicine and Health Sciences, contributing to the development of innovative initiatives with a high scientific impact.

The CHAMPS project “Characterization of amyloid pattern, structures and interaction with host’s cells using innovative multiphoton and LC-MS based technologies”, is part of the MYNESIS programme “A multiscale integrated approach to the study of the nervous system in health and disease”. The University of Molise is actively participating in the project by contributing to the structural and conformational characterisation of amyloid and amyloid-like aggregates, with a particular focus on their interaction with host cells.

The main objective is to investigate the biochemical and morphological properties of these aggregates through a multi-parametric strategy, based on the integration of two advanced technologies: FLIM/STORM microscopy in two-photon (2P) configuration for high temporal and spatial resolution imaging, and triple quadrupole LC-MS/MS mass spectrometry for quantitative analysis. The CHAMPS cascade project “Characterization of amyloid pattern, structures and interaction with host’s cells using innovative multiphoton and LC-MS based technologies” is part of the MYNESIS programme “A multiscale integrated approach to the study of the nervous system in health and disease”. The University of Molise is actively participating in the project by contributing to the structural and conformational characterisation of amyloid and amyloid-like aggregates, with a special focus on their interaction with host cells. The focus is on understanding the dynamics of protein aggregation, a key process in numerous neurodegenerative diseases.

This technological synergy makes it possible to correlate alterations in fluorescent lifetimes with aggregate morphology and measured molecular profiles. The expected results will help elucidate early mechanisms of amyloid aggregation and provide a platform for testing potential therapeutic agents.

Link: https://mnesys.eu/
Project code: PE000006
CUP: D33C22001340002
Funding: The project is financed by the European Union – Next Generation EU‘ on the PNRR MUR funds ’Mission 4 Component 2‘ ’From research to enterprise‘ Investment 1.3 ’Creation of partnerships extended to universities, research centres, companies for the financing of basic research projects”.

As part of the PRIN 2022 PNRR programme, the project “Investigation of common biochemical pathways, centered around the Amyloid precursor protein (APP), in Alzheimer’s Disease and breast cancer models: possible role of Purine Nucleoside Phosphorylase (PNP)”,

Nell’ambito del programma PRIN 2022 PNRR, il progetto “Investigation of common biochemical pathways, centered around the Amyloid precursor protein (APP), in Alzheimer’s Disease and breast cancer models: possible role of Purine Nucleoside Phosphorylase (PNP)”,

investigates the molecular mechanisms shared between Alzheimer’s disease (AD) and breast cancer (BC). The focus is on the interplay between APP, Grb2/ShcA and PNP, to elucidate their role in EGFR- and Notch-dependent amyloidosis and proliferation processes, as well as in the formation and release of extracellular vesicles (EVs). The aim is to shed light on the pathophysiological mechanisms of Aβ formation and cell cycle regulation.

The project applies advanced methodologies such as cross-linked mass spectrometry (XL-MS) to characterise interactions between key proteins and time-lapse and split-GFP fluorescence microscopy to study the dynamics of molecular interactions between proteins in real time. In parallel, structure- and ligand-guided virtual screening campaigns, supported by machine learning, aim to identify novel γ-secretase and Notch1 inhibitors capable of modulating the APP-Grb2/ShcA-PNP axis.

An innovative area of the project concerns the analysis of extracellular vesicles from cellular models that are characterised by LC-MS/MS proteomics and Western blotting, including APP, its fragments (C99, Aβ), PNP, Grb2/ShcA, EGFR, Notch1 and LRP8. Their functional effects are tested on 2D and 3D cell models using the xCELLigence system to monitor cell proliferation, death and invasiveness in real time.

Overall, the project contributes to the understanding of common molecular circuits between neurodegeneration and oncology, promoting the development of shared biomarkers and therapeutic targets through a translational approach.

Project code: P2022KPMB9-LS1
CUP: H53D23011120001
Funding: The project is financed by the European Union – Next Generation EU within the framework of the National Recovery and Resilience Plan (PNRR), Mission 4 “Education and Research”, Component 2 “From research to enterprise”, Investment 1.1 “Research Projects of Significant National Interest (PRIN)”.

The cascade project“FREEMODE-RNA – Fast, Reliable, Economic and Ethic Models to Study Delivery of Nucleic Acid-based Therapeutics”

is part of the spoke 8 “Platform for DNA/RNA delivery” of the National Centre for RNA & Gene Therapy.

The project aims at evaluating the efficacy, specificity and safety of delivery of therapeutic nucleic acids (RNAi, gRNA for CRISPR/Cas9, miRNA, antisense RNA, circRNA, aptamers) via nanocarriers and engineered nanovesicles. The animal models used are Drosophila melanogaster and Caenorhabditis elegans, chosen for their cost-effectiveness, speed of reproduction and ease of genetic manipulation. These model systems represent an ethical and sustainable alternative to the use of mammals, in line with the principles of the 3Rs (Replacement, Reduction, Refinement).

The main objectives of the project are (1) to verify and quantify the efficacy of nucleic acid uptake in in vivo models and (2) to assess their biological activity and potential toxicity, monitoring possible metabolic and off-target alterations.

To achieve these goals, advanced technologies are adopted: LC-MS/MS mass spectrometry and HPLC for metabolic-proteomic analysis, Western blotting for quantification of molecular targets, and high-resolution fluorescent imaging to investigate the intracellular distribution of nanocarriers. In addition, computational techniques and Artificial Intelligence algorithms are employed for the automated analysis of phenotypic and molecular data.

The project’s integrated approach enables rapid and efficient screening of the most effective and least toxic vectors for use in preclinical studies in selected mouse models. FREEMODE-RNA thus contributes to the rationalisation of the RNA drug development process, optimising resources and reducing the time and costs of the pre-clinical phase.

Link: https://www.rna-genetherapy.eu/it/
Project code: CN000041
CUP: E63C220009400071
Funding: The project is financed under the National Recovery and Resilience Plan (PNRR), Mission 4 “Education and Research”, Component 2 “From research to enterprise”, investment line 1.4 “Strengthening research structures and creating national R&D champions on certain Key Enabling Technologies”.

D3 4Health – Digital Driven Diagnostics, prognostics and therapeutics for Sustainable Health care is configured as an innovative health ecosystem composed of 28 partners including public and private universities, research institutes and enterprises, and carries out activities to enhance research on digital technologies in the health sector, through a sophisticated data mining process, in order to improve diagnosis, monitoring and treatment.

Within this initiative, the University of Molise plays a fundamental role within Spoke 1 from a scientific point of view, contributing significantly to the integration of innovative solutions based on Artificial Intelligence (AI) in clinical practice. The main objectives of the project include the development of algorithms and the validation of AI models aimed at predictive diagnostics and optimisation of therapeutic efficacy in four different pathological areas: central nervous system tumours, metastatic carcinoma of the rectum, liver cancer and multiple sclerosis.

Within Spoke 3, the University is co-leading and participating in the creation of advanced predictive models for personalised medicine through the creation of a Digital Twin, focusing its activities on three highly complex diseases: metastatic colon cancer, cholangiocarcinoma and liver cancer. The main activities include the identification and validation of new pathological biomarkers through retrospective and prospective studies, the characterisation of biological and physico-chemical markers from liquid biopsy, and the development of wearable sensors for monitoring oncological pathologies. Liquid biopsy represents an advanced and minimally invasive technology for the molecular characterisation of tumours through the analysis of components released by neoplastic cells into biological fluids, particularly blood. Among these, exosomes play a central role: they are nanometric extracellular vesicles actively secreted by tumour cells, containing proteins, lipids, mRNA, microRNA and double-stranded DNA. The content of exosomes closely reflects the molecular state of the cell of origin, making them extremely sensitive tools for the identification of diagnostic, prognostic and predictive biomarkers. Their stability in body fluids, combined with their ability to cross biological barriers, facilitates their use in dynamic disease monitoring, therapeutic response assessment and patient stratification.

The analysis of these elements can allow real-time assessment of the molecular evolution of the tumour, supporting early identification of recurrences, selection of targeted therapies and monitoring of therapeutic efficacy. Activities focus on the optimisation of extraction and analysis techniques, as well as the validation of multi-analyte panels that can be integrated into predictive digital models.

Link: https://sites.google.com/uniroma1.it/d3forhealth/home
Project code: PNC0000001
CUP: B83C22006120001
Funding: The project is financed within the framework of the National Plan for Complementary Investments (PNC) to the PNRR, with funds from the European Union – NextGenerationEU.

EU-funded HEALITALIA (Health Extended Alliance for Learning and Investigation through AI) – “NextGenerationEU” is an initiative funded by the Ministry of University and Research within the framework of the National Recovery and Resilience Plan (PNRR), which aims to develop and implement advanced solutions for precision medicine through the integration of clinical, omics and digital data. The network involves Italian universities, research centres and healthcare facilities, promoting a collaborative and interdisciplinary approach to improve the diagnosis, prognosis and personalised therapy of major chronic and complex diseases.

As part of Spoke 8 of the HEAL Italy initiative, the University of Molise is contributing to the development and implementation of precision medicine approaches, with particular reference to cardiometabolic disorders. The activity is part of the project “MOCCARDIS, Multi-omics characterisation of human samples derived from cardiometabolic disorders affected patients”, which involves the multi-omics characterisation of biological samples derived from patients affected by cardiovascular and metabolic disorders, through the integration of genomic, transcriptomic, proteomic and metabolomic data. A central element is the use of liquid biopsy for non-invasive analysis of the molecular processes underlying the disease.

In particular, the focus is on exosomes, extracellular vesicles released into circulation by pathological cells, which contain a rich and specific information load – including proteins, mRNA and microRNA – representative of the functional and metabolic state of the cell of origin. Through advanced isolation and analysis technologies, exosomes are being studied as potential biomarkers for early diagnosis, risk stratification and monitoring of therapeutic response in patients with diabetes, cardiovascular, liver and associated neurological diseases. The data obtained are integrated using interactive bioinformatics tools within a dedicated platform to support the construction of predictive models and the definition of customised clinical intervention strategies.

Link: https://www.healitalia.eu/en/
Project code: PE_00000019
CUP: I53C22001440006
Funding: The project is financed within the framework of the National Recovery and Resilience Plan (NRP), Mission 4, Component 2, Investment 1.3.The data obtained are integrated by means of interactive bioinformatics tools within a dedicated platform, to support the construction of predictive models and the definition of customised clinical intervention strategies.

The Inter-Universities Consortium for Engineering and Medicine (COIIM) was set up among the following Universities: University of Molise, University of Sannio and University of Cassino and Southern Lazio.