Bone is a dynamic tissue, which is constantly remodeled throughout life. Small shifts between bone formation and bone resorption lead to pathological situations. In osteoporosis and cancer-induced osteolysis, the excessive degradation of bone is due to increased osteoclast activity. The host institution uses target discovery to identify genes that are required in osteoclast function. Target discovery is a new interdisciplinary discipline involving molecular biology, bioinformatics, histology and cell biology. The host institution has already identified a series of genes that are up regulated during the formation or human osteoclasts in vitro. The aim of the present project is to establish the in viva expression of osteoclastgenes, and to assess their functional importance. The fulfillment of this aim requires a number of steps as explained in Form B4 (detailed proposal information: 3.1.2). More particularly, I will take care of the following tasks as integrated parts of the target discovery program.
(1) I will use laser micro dissection microscopy to isolate single osteoclasts from human biopsies collected by the host institution, as well as from animal models for osteoporosis and bone metastasis currently used in the host institution.
(2) I will use quantitative PCR to quantify accurately the expression of the genes that my colleagues will have selected through large scale gene profiling, bioinformatics and tissue arrays.
(3) I will perform functional validation by using antisense, function blocking antibodies or viral transfection of osteoclasts, in different models that allow to evaluate the osteoclastic resortive activity and that have been developed in the host institution. The project is an opportunity to learn to work in a multidisciplinary program. I will become an important and integrated member of an established group of experts in different scientific areas, and will get immediate access to a large number of highly innovative and advanced techniques. More specifically, I will be involved in high-level technologies such as laser micro-dissection microscopy, quantitative PCR, osteoclast transfection, and frontline bone biology, I will have to coordinate my work with my colleagues, and adapt my studies according to information arising from large scale gene expression profiling, bioinformatics, tissue arrays, in site hybridizations, I will have a demanding scientific responsibility for planning, designing, execution and conclusion of the experiments required by the multidisciplinary program. My work will help to bring the information the host institution got on regulated genes, to a mature level. My work will lead to the long-term opportunities of Nordic Bioscience to develop new drugs for treatment of osteoporosis and bone metastasis.