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Role of ion channels in lymphocyte development and in leukemia clonal evolution
Universita degli studi di Firenze
Experimental and Clinical Medicine Department (General Pathology Section)
Type of Research Project
- Basic science
What is the background of the project?
The project is based on the phenotypic characterization of the Erg1b KO transgenic mouse model. Lymphocyte’s precursors will be isolated from the principal lymphoid organs (bone marrow, thymus, spleen, blood) and different signalling pathways interrogated. Acute lymphoblastic leukemia is the most common malignancy of childhood, long-term survival rates being still low due to systemic toxicity and chemoresistance. In the Hosting Laboratory, it has been provided evidence that different leukemias like BCP-ALL, where B-cell development is arrested at the pre-B checkpoint, and T-ALL overexpress a peculiar isoform of the ether-a-gó-gó-related gene 1 (hERG1), the so called hERG1B encoding for a K+ channel. In both AML and T-ALL, hERG1B controls relevant aspect of leukemia establishment and progression and its expression correlates with a worse prognosis. IFMSA International Secretariat, c/o IMCC, Nørre Allé 14, 2200 København N., Denmark The current hypothesis is that these facts can be traced back to a peculiar and relevant role exerted by hERG1B’s activity at specific stages of either B cell and T cell development.
What is the aim of the project?
The main aim of the present proposal is to unravel the mechanisms underlying hERG1b over-expression and dysregulation in B-ALL and T-ALL, in order to define the role and the function of the channel in the leukemogenic process.
What techniques and methods are used?
The process of B and T lymphocyte development and the signalling pathways that regulate it will be characterized using cytofluorimetric analysis techniques. The above mentioned antibodies will be developed applying molecular biology techniques, such as PCR, affinity chromatography, cloning, bacterial cells manipulation, trasfection, tumor cell culture, antibody labelling with fluorophores.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
At the beginning the student will follow the tutor during the experiments. As he/she has acquired expertise will perform the experiments for: - Isolation of lymphocytes and precursors from the principal murine lymphoid organs. - Phenotypic characterization of the isolated cells using cytofluorimetric techniques. - Check the different signalling pathways that regulate lymphocyte development. This will be performed by western blot techniques and by cytofluorimetric analysis.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
To understand the role of ion channels during lymphocyte development It is recommended the reading of the suggested papers. The rest of the teaching will be provided by the tutor during the course of the internship.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Good level of English, to be able to read scientific papers. Laboratory expertise will be acquired on site.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- - Feske S; Skolnik EY; Prakriya M. Ion channels and transporters in lymphocyte function and immunity. Nat Rev Immunol. 2012 Jun 15;12(7):532-47.
- - Arcangeli A; Crociani O; Lastraioli E; Masi A; Pillozzi S; Becchetti A. Targeting ion channels in cancer: a novel frontier in antineoplastic therapy. Curr Med Chem. 2009;16(1):66-93. Review. PMID: 19149563
- - Pillozzi S; Masselli M; DeLorenzo E; Accordi B; Cilia E; Crociani O; Amedei A; Veltroni M; D’Amico M; Basso G; Becchetti A; Campana D and Arcangeli A. Chemotherapy resistance in acute lymphoblastic leukemia requires hERG1 channels and is overcome by hERG1 blockers. Blood. 2011 Jan 20;117(3):902-14. doi: 10.1182/blood-2010-01-262691. PMID: 21048156
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