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Role of IsomiR in the pathogenesis of colorectal cancer: from miRNome-wide bioinformatic analysis to their functional validation
Lithuanian University of Health Sciences
Institute for Digestive Research
Prof. Limas Kupčinskas
Dr. Jurgita Skiecevičienė; Dr. Violeta Šaltenienė
Type of Research Project
- Basic science
What is the background of the project?
Colorectal cancer (CRC) is one of the most prevalent cancer types worldwide and is a leading cause of cancer related mortality. Despite increasing availability of CRC screening programs, more precise methods for early non-invasive disease diagnostics are highly needed. The biggest breakthrough in the personalized management of malignant diseases is expected in the field of molecular biomarkers. Currently, microRNAs (miRNAs) are one of the most widely studied molecular biomarkers in oncology. Previous studies have revealed that deregulation of miRNAs occurs virtually in all major types of cancer, including CRC and is associated with disease initiation, progression and metastasis. Recent studies revealed that archetypal miRNA sequences can have various isoforms (isomiRs), which may have distinct expression patterns from archetypal miRNAs or target different mRNAs. By studying only archetypal miRNAs, researchers miss a high number of possible disease related associations. Overall, our knowledge about isomiRs is extremely limited. To date, these molecules have been studied only in the breast cancer, melanoma and gastric cancer groups, but no previous study on CRC has been performed.
What is the aim of the project?
Therefore, this study would be the first in the world which is aimed to investigate deregulation of isomiRs in CRC and define their target genes and function in the oncogenesis.
What techniques and methods are used?
1. Evaluation of changes in cellular processes (proliferation, apoptosis, migration and etc.) of cancer cells after transfection with selected isomiR mimics or inhibitors. 2. Experimental validation of isomiR target genes using RT-qPCR, Western Blot and Luciferase Reporter Assay.
What is the role of the student?
- The student will mainly observe
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Week 1: getting familiar with the different methods employed in the laboratory; Week 2-4: work with different cell lines; transfection methods; functional methods (MTT, wound-healing assay, colony formation etc.); qPCR of selected target genes; luciferase assay
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will acquire detailed instructions and protocols of the methods used in the laboratory during the 1st week of the traineeship. Daily structured plan and protocols will be provided to the student. Close supervision, as well as daily discussions will be scheduled. At the end of the traineeship a final report has to be submitted to the head of the laboratory.
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 scientific report - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
We expect that the student will attend at least 80% of the research exchange, be responsible for his/her actions at the practice and be accountable to the supervisor and LORE/NORE.
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) - Students in biomedical fields
- Telonis AG et al. Knowledge about the presence or absence of miRNA isoforms (isomiRs) can successfully discriminate amongst 32 TCGA cancer types.Nucleic Acids Res. 2017 Apr 7;45(6):2973-2985. doi: 10.1093/nar/gkx082.
- Telonis AG et al. Beyond the one-locus-one-miRNA paradigm: microRNA isoforms enable deeper insights into breast cancer heterogeneity.Nucleic Acids Res. 2015 Oct 30;43(19):9158-75. doi: 10.1093/nar/gkv922. Epub 2015 Sep 22.
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