Multidimensional analysis of the tumor microenvironment of cholangiocarcinoma
The immune tumor microenvironment (TIME) consists of different cell types that can modulate the tumor-directed immune activity and influence the response to therapy. Cancer-associated fibroblasts (CAFs) represent the dominant non-hematopoietic cell type within the TIME of many cancer types, including biliary tract cancers. CAFs are pivotal in tumorigenesis, tumor progression, and metastasis. Additionally, CAFs are actively involved in immune regulation by producing inflammatory cytokines/chemokines and soluble factors. Nonetheless, the exact mechanisms that define how CAFs shape the TIME of cholangiocarcinoma (CCA) remain underexplored. Thus, the overall goal of this study is to generate the first in-depth transcriptional map of the tumor-immune microenvironment of CCA and decipher the molecular mechanisms through which CAFs influence its immune-tumor landscape.  Targeting these aberrant pathways within stromal cells may increase response rates to immunotherapy and will have important implications for the design of innovative combination strategies.

Identification of novel therapeutic approaches for patients with biliary tract cancer
Biliary tract cancers (BTC) represent a heterogeneous group of malignancies with relatively low-incidence and poor prognosis. Therapeutic options for BTC patients at advanced stage are severely limited and no molecular targeted therapy has been approved yet for this diseases.
It is known that tumor and immune cells fuel tumor angiogenesis and vice versa. Understanding the intricate relationship underpinning this process is crucial for the design of therapeutic approaches. We will generate ad-hoc 3D spheroids which are superior to 2D monolayer cell cultures in mimicking the interaction between supporting cells and tumors cells. The 3D systems herein generated will include custom 3D spheroid cell culture and more complex organoids which will represent a powerful platform for high-throughput screening of FDA-approved drugs. The results will allow the identification of potential effective treatments whose efficacy will be then validated, as single or combination regimens, in a variety of animal models of the disease.

Epigenomic profiling of cholangiocarcinoma
Aberrant methylation is widely recognized as a key feature of human cancers. A growing body of evidence suggests a role for epigenetic modulators in influencing the tumor microenvironment and the intra-tumor immune response. Nonetheless, a comprehensive deconstruction of the methylation profile of biliary tract cancers is lacking. In order to fill this knowledge gap, we will evaluate epigenetic dysregulation associated with DNA methylation in a large cohort of cholangiocarcinoma patients. The analysis will ultimately provide novel insights into the pathogenesis of the disease and uncover candidate somatic epigenetic alterations driving tumor growth.

Integrative molecular analysis of gallbladder cancer
Gallbladder cancer remains significantly underexplored compared to the other tumors of the biliary tract. Furthermore, to date no molecular classification has been generated for this disease. We will perform the first multi-omics analysis through integration of whole-genome expression with DNA methylation, mutations and chromosomal aberrations in a large human cohort. Novel candidate therapeutic targets herein identified will be then validated in ad-hoc in vitro and in vivo models of the disease (i.e. 3D organoids, patient-derived xenografts, etc).