INNOVAtive tissue and non-invasive biomarkers for a precision medicine in CHOLANGIOcarcinoma: early diagnosis and therapeutic response of tumor subtypes. INNOVACHOLANGIO

Cholangiocarcinoma (CCA) has recently been reclassified from anatomical and histological point of views. In particular, two distinct histological types of intrahepatic CCA are recognized: small bile duct and large bile duct; the latter can be assimilated by histo-morphological characteristics to hilar/perihilar and distal CCA. The distinct intrahepatic CCAs have different clinical-pathological characteristics, prognosis, molecular profile, and etiopathogenesis. In view of the complete innovativeness of this classification, which has been approved by both ICD-11 and ICD-0 3.2, in operation since beginning of 2022, a comparative analysis of specific biomarkers for early diagnosis and drug responsiveness of the two tumor subtypes is urgently needed. The main aim of INNOVACHOLANGIO project is to leverage enhanced biomarker discovery and spatial single-cell technology to tackle the diagnostic delay in the diagnosis of CCA and providing personalized treatment based on accurate histo-morphological classification and biological clusters. In this project, enrolled subjects will undergo deep clinically and radiologically phenotyping (WP#1). In WP#2 the tissues (neoplastic, peritumoral, non-neopalstic liver diseases, and normal liver by donors) will be analyzed trough a single-cell definition and anatomic precision by the innovative Nanostring technology, allowing the histo-morphological classification and definition of homogenous biological clusters. These will be correlated with clinical and radiological parameters captured in WP#1, and enable the identification of diagnostic and prognostic biomarkers and targetable molecular pathways (tissue proteomic and transcriptomic-based classification of CCA). In WP#3, non-invasive multiparametric algorithms, based on clinical/radiological data plus protein- or metabolite-based liquid biopsy, will be developed for the accurate prediction of the histological subtypes and biological clusters (comprising therapeutic targets) of CCA. WP#4 will be focused on the clinical translation: validation in a prospective, multicentre cohort of non-invasive multiparametric algorithms for early diagnosis of CCA in high-risk patients, e.g. subjects with new diagnosis of biliary stricture or dominant stricture in primary sclerosing cholangitis, or hepatic nodules in liver cirrhosis, or subjects undergoing curative resection for CCA. The prospective validation will be carried out in a clinical context characterized by high risk of CCA; In these subjects, after appropriate clustering, biomarkers and algorithms for the specific tumor type will be tested in a prospective multicentre longitudinal study. The accurate definition of the specific tumoral features and biomarkers in the defined population of subjects at risk (e.g. small bile duct iCCA in subjects with liver cirrhosis, or large bile duct iCCA and pCCA in sclerosing cholangitis) will enormously facilitate early diagnosis, and the definition of prognosis and therapeutic response.