WP1: A hybrid Scattering/Raman microscope was developed and characterized, and an imaging protocol was designed to enable coregistered measurements with the existing at HMGU OA/SHG/THG microscope. In specifics, RiverD developed a Raman module, optimized for tissue Raman microscopy in the red and near-infrared region of The spectrally resolved depolarization spectra module was developed by UC3M. the electromagnetic spectrum. The control and data pre-processing software was programmed in MATLAB and C#, using a .NET framework, while a software developer kit (SDK) was created by RiverD and RAYFOS for the software of the hybrid microscope.In collaboration with RiverD, UC3M designed the coupling interface of the scattering module for straightforward coupling to the hybrid microscope. A LabVIEW based software was also developed. The integration of these two modalities into a new hybrid microscope was implemented by HMGU in close collaboration with RiverD, UC3M, and RAYFOS. The microscope was successfully is WP3.
WP2: Experiments were performed towards the development of mouse models in one hand, and establishing procedures for obtaining human tissues and actual collection of the first biopsies. Two different mouse models for esophageal and colorectal carcinoma have been developed. A small scale breeding program is running at HMGU. This was necessitated by the need for measurements on fresh material. The esophageal model which is going to be measured relies on the overexpression of the IL2 cytokine which is known to lead to inflammation, hyperplasia and eventually metaplasia and cancer. The colorectal model relies on the tissue specific knockout of the tumor suppressor protein APC in the small and large intestine of mice. With respect to human samples, protocols for taking biopsies from patients have been established. Ethical approval for patient work has been obtained from the independent review board (IRB) of the UMCG. First biopsies (n = 32) have already been acquired from patients with Barrett’s esophagus and Lynch syndrome in the first 2 weeks since we are running, and more patients are scheduled.
WP3: The optical measurements with the Scattering/Raman and the hybrid SHG/OA/2PEF microscopes on mouse and human tissue samples were performed by HMGU. Unique pattern fields were extracted and used in correlation studies with the molecular data which were generated in WP4. Squamocolumnar junction, forestomach and stomach from over 30 control and experimental mice were measured. For the intestine, tumor and nontumor mucosa from APC knockout mice were measured. As controls, we used healthy mucosa from controlo miced. Fixed esophageal and colorectal biopsies from Barrett's and Lynch synderomw patients were meaused alongside controls.
WP4: The molecular profile of all biopsies in WP3 were molecularlyh characterized with RNA-seq using the Illumina platform at BRFAA. RNA-seq data were processed and deregulated cell processes were identified. Interestingly, the molecular profiles and optical measurement seem to correlate significantly. Moreover, tissue adjacent to the tumor, albeit histologically normal, seems to share similarities with both the tumor and the control mouse counterpart.
WP5. Two clinical trials were performed on Barrett's and Lynch syndrome patients. A total of 30 patients have been measured so far with very promising results.
