Minimal Disease: Molecular-orientated early diagnosis of colorectal and neuroendocrine tumors (focus: small cell lung cancer)

Currently, for solid malignancies there is no routinely established molecular-based method to detect minimal disease (MD), which describes a malignant disease in its early stages before clinical manifestation. There is strong need for molecular-oriented research to detect minimal disease since no other methodology is likely to provide the necessary sensitivity.

Aim
The aim of this project is to define clinically useful molecular-orientated early detection of minimal disease in colorectal, lung, prostate and ovarian cancer. “Methylation Signatures” that signal the presence of MD will be investigated at the epigenomic level and in a broad spectrum of biological materials (stool, sputum, alveolar lavage, plasma). The significance of this kind of “extended diagnostics” will be assessed.

Pre-existing knowledge
This project is based partly on the findings of a currently ongoing collaborative project of the LBI for Gynecology and Gynecologic oncology. The project focuses on the selection of epigenetic markers for the detection and/or analysis of ovarian cancer cells. So far hypermethylation of the gene N33 is a candidate marker fro early diagnosis of ovarian cancer. The methods for the analysis of hypermethylation in free DNA in serum/plasma are well established.
Further, commercially available test kits and pre-release version of test systems in development for detecting hypermethylation of selected genes in stool, sputum/alveolar lavage, and urine/ejaculate for the early detection of colorectal, lung, and prostate cancer, respectively, will be available free of charge from OncoLab Diagnostics GmbH. in the scope of a scientific collaboration.
There is access to all necessary lab equipment and infrastructure.

Methods
Stool, sputum (and alveolar lavage if accessible), and serum/plasma samples will be collected from cancer patients and healthy volunteers for control. Specimens will be processed according the instructions of the manufacturer. DNA will be isolated with the appropriate isolation reagents (also provided) and chemically modified by bisulphite treatment. Hypermethylation of selected genes will be analysed by methylation-specific PCR and methylation-specific reverse hybridization.

Workplan
Biological specimens shall be collected throughout the duration of the project (36 months). Numbers of patients to be included in the study will only be limited by the number of available patients. However, minimum sample numbers necessary to reach statistically significant results will be calculated by a statistician before starting the project. Sample analyses will be performed continuously, or at certain time points, if suitable.
Final results will describe the significance (sensitivity, specificity) of single hypermethylation markers/marker combinations and their frequencies.