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Validation of a microfluidic electrophoresis method to detect microsatellite instability in tumors
Spain (IFMSA Spain) - University of La Laguna, Santa Cruz de Tenerife
Anatomía Patológica, Facultad Medicina, Ofra s/n, La Laguna 38320 Tenerife, Spain
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Microsatellite instability (MSI) is a marker of about 15% of colon cancers, a subset of tumors in which one of the genes involved in mismatch DNA repair (MMR genes, mainly MSH2, MSH6, MLH1 and PMS2) has suffered either somatic or germline mutation, or epigenetic inactivation. Detection of MSI is clinically relevant not only because it might help identify hereditary colorectal cancer (CRC) cases (Lynch syndrome) but also because MSI has important implications in CRC treatment. MSI is currently ascertained by either indirect methods (immunohistochemical detection of MMR protein products, MMR.ihc) or capillary electrophoresis of microsatellite amplicons (MSI.ce). In the clinical setting, both methods have limitations.
What is the aim of the project?
We want to validate the use of microfluidic, high resolution electrophoresis to test MSI in CRC, comparing it with either MMR.ihc or MSI.ce
What techniques and methods are used?
DNA purification from tissue embedded in paraffin PCR amplification of microsatellites Standard electrophoresis to ascertain amplifications High resolution microfluidic electrophoresis Capillary electrophoresis Immunohistochemistry IFMSA International Secretariat, c/o IMCC, Nørre Allé 14, 2200 København N., Denmark in general in this project?) This is an exploratory, feasibility project. We do not plan to perform statistical analysis, in the case of doing it with the student during his stay, we will prepare contingency tables comparing two techniques: ihc and MSI and Chi square / Fisher exact tests will be used to assess the differences in percentages found.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will learn and perform purification of DNA from tissue embedded in paraffin. Also, to amplify DNA through PCR amplification of microsatellites, to detect DNA amplifications by standard electrophoresis, high resolution microfluidic electrophoresis, capillary electrophoresis; also perform immunohistochemical techniques. The student will be involved in the analysis of results, either qualitative (if we just process a few samples of known genotype) or after statistical analysis This is an exploratory, feasibility project. The IFMSA International Secretariat, c/o IMCC, Nørre Allé 14, 2200 København N., Denmark students typically have time to only learn the techniques and process a few controls (samples that we know they have / have not microsatellite instability). We do not plan to perform statistical analysis. In the hypothetical case that the student already knows how to work in the lab and more than 10 samples are studied, we will prepare contingency tables comparing two techniques: ihc and MSI and Chi square / Fisher exact tests will be used to assess the differences in percentages found.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Four days a week, 30min. research seminars given by staff members of the dept. pathology
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Good work ethics
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- Hajirawala L; Barton JS. Diagnosis and Management of Lynch Syndrome. Dis Colon Rectum. 2019 Apr;62(4):403-405. doi: 10.1097/DCR.0000000000001345. PubMed PMID: 30844969.
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