Info​
 

Tumors: The project aims to optimize the relevant protocols for assessing the MechanoOptical signatures on fresh tissue biopsies from breast cancer, which is one of the most highly desmoplastic tumors. Breast cancer is the most common cancer in women worldwide, is a complex cancer type with heterogeneous clinical behavior and response to therapy. The project will focus on triple negative breast cancer, that is a unique subgroup with poor prognosis. Therefore, there is an urgent need to better understand the clinical behavior of breast cancer by developing and using targeted predictive biomarkers that will lead to patient-specific treatments.

■RO1: We will carry out in vivo studies in two orthotopic tumor models. During cancer progression, biopsies with automated biopsy guns will be obtained at different time points. The same day of tissue harvest, the tissue biopsies will be embedded in agarose and subsequently will be sectioned with a vibratome. AFM and fluorescence microscopy (after standard staining protocols) will be performed in order to evaluate the sectioning performance. Expected results: optimize all the steps of the protocol for sectioning fresh tissue biopsies. The aim is the develop protocol to offer appropriate sections for both AFM characterization and fluorescence intensity measurements.

■RO2: The aim of this RO is to develop the appropriate protocol so to assess collagen-based fluorescence intensity from the biopsies that will be derived from the RO1. The challenge is to use low-cost, fiber-based techniques for assessing collagen and cell nuclei fluorescence from fresh tissue sections (without any fixation) that are mounted on the AFM system. Tissue biopsies will be mounted on poly-L-lysine-coated AFM-compatible glass bottom dish. Subsequently, tissue collagen and cell nuclei will be stained and the dish will be mounted on AFM system. The final aim is to integrate a dual-wavelength fluorescence system for simultaneous measurements of the fluorescence intensity of collagen and cell nuclei. Also, immunofluorescence of tissue sections will be performed with fluorescence microscopy so as to evaluate the staining procedure. Expected results: Develop a protocol for staining and measuring fluorescence intensity from fresh tissue sections mounted on AFM system.

■RO3: The results of the two previous ROs will be used. The aim is to achieve the simultaneous AFM and fluorescence intensity measurements on fresh tissue specimens. For this RO we will develop the same murine tumor models as in RO1. Then we will employ chemotherapy (doxorubicin), anti-fibrotic treatment (tranilast) and the combination of the two treaments (chemotherapy + tranilast). Biopsies with automated biopsy guns will obtain at different time points. The biopsies will be used for assessing the MechanoOptical signatures, while immunofluorescence and histological analysis will take place with classical optical microscopy techniques. Expected results: Optimize protocol for simultaneously measurements of nanomechanical and fluorescence properties of fresh tissue specimens