Investigation of Cancer Cell Extravasation Through mRNA Analysis of Organ-specific Endothelial Cells and Microfluidics
Condition: Breast Cancer Metastatic
Study type: Observational

Summary: 
      The project aims at unraveling the role of organ-specific endothelia mediating the
      preferential metastasisation of breast cancer cells to bone by using a multi-faceted
      approach, integrating microfluidics and transcriptomic profiling. Based on a recently study
      published by the investigators [Jeon et al., PNAS 2015], it can be hypothesized that
      phenotypic differences at the level of organ-specific endothelial cells are able to drive the
      preferential extravasation of breast cancer cells to specific sites. Hence, the
      transcriptional profile of primary organ-specific endothelial cells derived from healthy
      patients (i.e. non-affected by breast cancer) will be analyzed to identify phenotypic
      differences between organ-specific populations of endothelial cells. These analyses will
      allow to identify potential target genes involved in the organ-specific extravasation of
      cancer cells (i.e. genes differentially expressed by endothelia of preferential and
      non-preferential metastasisation sites). The selected genes will be silenced and the effect
      of gene silencing will be evaluated through microfluidic in vitro organ-specific 3D models
      designed to study cancer cell extravasation.
    

Detailed description: 
      In particular, the main aim of the study will be to highlight differences between bone and
      skeletal muscle microenvironments, which are respectively preferential and non-preferential
      metastasisation sites for breast cancer cells, in order to identify specific pathways driving
      breast cancer cells extravasation. To this purpose, differences in the transcriptional
      profile of ECs derived from bone and muscle endothelium will be investigated. These analyses
      will be used to select target genes differentially expressed by bone- and muscle-specific
      ECs. Then, ECs obtained from bone and muscle endothelium will be respectively used to mimic
      bone and muscle microvessel environments in microfluidic in vitro 3D models allowing for the
      study of breast cancer cell extravasation. The genes selected according to the results of the
      transcriptomic analysis (e.g. genes expressed in ECs derived from bone endothelium, but not
      expressed in ECs derived from muscle endothelium) will be silenced and the effect of gene
      silencing will be evaluated monitoring breast cancer cell extravasation in order to verify
      the involvement of the selected genes in this process.
    

Intervention name(s): Tissue samples collection and cells isolation
Intervention type(s): Other
Intervention description(s): During surgery, leftover samples of bone (femoral head/ trabecular bone tissue) and muscle will be collected. Then they will be sent to the laboratory where isolation of cells will be performed.
Observational model: Other
Time perspective: Prospective
Sampling method: Non-Probability Sample
Study population: 
        Patients undergoing hip or knee surgery, from whom samples of waste bone and muscle tissues
        can be harvested.
      
Biospecimen description: 
      Samples of bone and muscle tissue
    
Keywords: Organ specific endothelium; Microfluidics; Cancer cell extravasation