A high-throughput microphysiological platform to analyze the role of the aging human vasculature in melanoma metastases
Description: The effective treatment of metastatic melanoma is an unmet clinical need since less than 50% of patients survive at 5 years after diagnosis of advanced disease. This dismal prognosis is mainly due to the formation of brain metastases for which no effective therapies are available. Of all cancer types, melanoma has the highest rate of cerebral tropism, with over 40% of advanced patients developing brain metastases.Clinical observations have shown that patients above 70 years old have metastatic melanomas with the most aggressive prognostic features, which result in the highest mortality rate. Surprisingly, the impact of physiological aging on the pattern of melanoma metastases is largely unexplored when studying the biological mechanisms driving cancer cell dissemination. Pre-clinical and clinical studies have shown that blood vessels and lymphatics experience structural and functional changes during physiological aging, including compromised cell-cell junctions and increased vascular permeability, particularly in the blood brain barrier (BBB). This compromised vascular function coupled with the loss of vascular-identity genes increases the transmigration of circulating melanoma cells in the brain (i.e. extravasation). However, a detailed mechanistic explanation of the higher rate of melanoma cell transmigration through the vasculature of old patients is still lacking, partially due to the absence of reliable models that:1) recapitulate features of vascular aging;2) allow high-throughput (HT) parametric studies to identify drivers of melanoma cell extravasation.To fill this gap, we propose to:1) design a high-throughput system hosting 3D perfusable human BBB minitissues;2) compare BBB models derived from human donors of different ages and validate them at the molecular and functional level by comparison with human brain tissue biopsies and published datasets;3) analyze the biological mechanisms driving the age-dependent extravasation of melanoma cells and provide a list of candidate targets involved in melanoma extravasation for functional perturbation;4) perform a CRISPR-based screening of the most promising targets and validate themA key aspect of the proposal is the multidisciplinary integration of microvascular network (MVN) biofabrication, multi-omic characterization of human brain vasculature, and design of a data analysis methodology that exploits artificial intelligence to combine experimental data with existing databases to identify and rank pathways underlying melanoma extravasation. During the project we then expect to:1) design and microfabricate a HT platform hosting human 3D minitissues that allows to perform large-scale experimental campaigns; this platform is fully integrated with HT biofabrication strategies and HT analytical techniques, and can find applications in any other field requiring screening of compounds or drugs (e.g. combination of chemotherapies on patient-derived cells);2) provide a detailed molecular and functional analysis of the differences characterizing the human BBB during aging, which can have consequences in other disease models including neurodegeneration;3) identify drivers of melanoma extravasation through the BBB by combining the molecular and functional profile of vasculature and cancer cells through HT live confocal imaging, automated quantification of local vascular permeability and cutting-edge spatial transcriptomics; the design of a novel data analysis pipeline is expected to discover potential mechanisms of extravasation also shared by immune cells during inflammatory processes or other phenomena involving immune cell mobilization;4) validate the identified targets through a functional CRISPR screening and human tissue microarrays, which provide a higher clinical relevance and are often neglected in basic biology studies.
Amount: CHF 800,000
Funding type: research
Funder scheme: Project funding
Start date: 2025-06-01
End date: 2029-05-31
Investigator: Bersini, Simone
Institution: Ente Ospedaliero Cantonale – EOC
Funded outputs count: 0
Funder award ID: 10002962
OpenAlex ID: G7863588857