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In Vitro characterization of lung cancer stem cells through cancer progression
Belgium (BeMSA) - KU Leuven, Leuven
Translational Cell & Tissue Research Dept. of Imaging and Pathology
Type of Research Project
- Basic science
What is the background of the project?
Investigation in lung cancer has been increased in the last decades, as this disease has become one of the most deadly diseases in the developed countries. However, it is still a poorly understood cancer and therapies used have only marginally improved the survival rates. Understanding of the cellular and molecular components of that behaviour is a main interest in current lung-cancer research. Our lab has previously described Lgr6 (Leucine Rich Repeat Containing G Protein-Coupled Receptor 6) as a lung stem-cell marker. In addition, we have observed the enrichment in Lgr6+ cells during lungadenocarcinoma progression. Lgr6+ cells harbor stem-cell properties (self-renewal and differentiation), and higher tumorigenic potential. The mechanism of selection comprises defective repression of the miR-17-92 cluster (microribonucleic acid-17-92 cluster). High levels of the miR-19 member target and reduced p38a-protein levels providing a specific survival signal for Lgr6+ cells, mediated by increased Wnt/b-catenin activity. We are now interested in a functional and molecular characterization of these Lgr6+ tumoral cells.
What is the aim of the project?
Application of cellular and molecular techniques to study the contribution of special cell types with stem cell properties to the initiation and development of Lung Cancer.
What techniques and methods are used?
Tumorigenicity, invasion and metastatic potential will be addressed and assessed in vitro and in vivo. Multiple in vitro techniques will be used such as soft agar growth, boyden chambers, ex-vivo tumor growth and stem cell differentiation (using mouse and human lung explants), and genetic expression profile of Lgr6+ cells from human lung tumors at different stages of cancer development. We have broad experience in all those techniques and the culture of multiple primary lung and tumor cells. - Isolation, culturing and assessment of cells extracted from human lung cancer tissue with different cell markers. - Comparison between cell populations of their transforming and cancer initiation potential. - Genetic comparative analysis of cell populations. - Genetic manipulation of target genes to test contribution to cancer
What is the role of the student?
- The tasks of the student will be performed on his/her own
What are the tasks expected to be accomplished by the student?
Learn and participate in the maintenance of lung stem cell populations in culture. Learn and perform DNA (deoxyribonucleic acid) and protein isolation and analysis by qPCR (quantitative polymerase chain reaction) and western blot. Participate in the analysis and discussion of the results. Learn and perform isolation and characterization of lung populations by Flow cytometry and microscopy.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary reading of articles, seminars.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student’s name will be mentioned in a future publication - The student will prepare an abstract
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Curiosity, good lab practice, working enthusiasm, intellectual contribution.
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
- Regulation of Human Lung Alveolar multipotent cells by a novel p38a MAPK/miR-17- 92 axis. F. Oeztuerk-Winder; A. Guinot; A. Ochalek and J-J. Ventura. EMBO J. 2012 Jul 24;31(16):3431-41
- A paracrine network regulates the crosstalk between human lung stem cells and the stroma. EJ. Ruiz; F. Oeztuerk-Winder and J-J. Ventura. Nat Comm. 5 (2014) Jan 16: 3175.
- miR-17-92/p38α dysregulation enhances Wnt signal and selects Lgr6+ cancer stem cell-like cells during human lung adenocarcinoma progression. A. Guinot; F. Oeztuerk-Winder and J-J. Ventura. Cancer Res. (2016) May 4; 76 (13); 1-11. doi: 10.1158/0008-5472.CAN-15-3302.
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