Medullary Thyroid Carcinoma (MTC) originates from mutations in calcitonin-producing parafollicular C cells of the thyroid, is a rare malignancy, accounting for 3-4% of all thyroid carcinomas. It occurs in a hereditary form (HMTC, 25%) or in a sporadic form (SMTC, 75%). The prognosis for patients with MTC is poor, as the tumor metastasizes at early stages; and the only curative therapeutic option so far is radical surgery. Genetic analysis helps identify inherited cases at a stage where prophylactic surgery can be offered to carriers of such mutations to prevent the disease. This approach may also be used to determine better treatment options for patients who are already diagnosed with MTC.
The goal of this project was to develop a comprehensive mutational panel for the detection of clinically relevant mutations in MTC samples. A total of 143 mutations (nucleic acid variations) in 8 human genes were selected from numerous papers and public databases and included into the MTC mutational panel. The selection criteria were based on the coding mutations (mutations that occur in the coding area of the genes) that were reported to occur in MTC patients and considered functionally relevant. Some of the mutations included into the MTC mutational panel (such as BRAF gene V600E mutation) were also described in other types of thyroid cancer, but most of the mutations were unique to MTC (such as all RET gene mutations). The entire assay design was carried out using Sequenom's online design tools (ProxSNP and PreXTEND (https://www.mysequenom.com/Tools) and Assay Design software (v. 3.1)).
The final file comprised from 115 assays corresponding to all 143 mutations included into the MTC panel. This file will be further processed using the SEQUENOM® Mass-ARRAY iPLEX® platform for DNA genotyping of clinical samples by the cancer research scientists at the Abramson Cancer Center of the University of Pennsylvania
Save for Anaplastic Thyroid Carcinoma (ATC), MTC is the second most aggressive type of thyroid cancer. Hereditary mutations that could lead to HMTC need to be found as rapidly as possible, as the onset age is only 5. This Mutational Panel is one of the best ways to diagnose any type of MTC, be it hereditary or sporadic, and give a small beam of hope to the poor person diagnosed with the mutation.
The decision to make this panel was a hard one, as at the time that I started it, I was going to take pre-calculus in Drexel University. It was an insane amount of work, but I managed to compile the list of mutations over the summer, and continue the analysis during autumn. The actually write-up was finished much later, when I decided that I could use that project as my capstone.
Simply put, the project was not easy. It took me a long time to understand the basics, and even a longer time to be able to manipulate small details, but in the end, the project was amazing. I have to say, that out of all the work I did while I was in SLA, this capstone project is the one I am proud of the most!