Comprehensive characterization of GWAS hits – pipeline to novel drug targets for anabolic treatment of osteoporosis (GWASforAna)

Osteoporosis is the most common skeletal disease with deterioration of bone structure and increased risk of bone fractures. Even though the biology of osteoporosis has been well studied, effective personalized approach to prevention, diagnosis and particular anabolic drugs to prevent bone fractures are still lacking and there is an urgent need to identify novel targets for anabolic treatment and prevention of bone fractures. One of the powerful approaches to identify novel drug targets is identification of genetic loci that are associated with osteoporosis phenotypes in genome-wide association studies (GWAS). GWAS result in hundreds of genetic loci bearing potential to be a drug target or biomarkers for prediction of fracture risks. Yet, to benefit from these big data, functional characterization of potential drug targets is necessary. Therefore, our objective is to set up a platform for functional characterization of genetic targets associated with bone diseases. In this project, we will thoroughly functionally characterize a set of GWAS hits using state-of the art methods including mesenchymal stem cells, genome editing by CRISPR/Cas9 techniques, in vivo animal model and analysis of clinical samples. Specifically, we will (A) first determine whether a selected gene target plays a role in bone formation by gain and loss of function experiments during osteogenic differentiation of human mesenchymal stem cells (MSC). The impact of target gene silencing on osteogenic differentiation will tell us whether or not a target gene plays a role in bone formation. Given the pleiotropic nature of targets, we will also examine the function of genetic targets during formation of adipocytes and miocytes. Similarly to osteogenic differentiation, MSCs will be genetically modified and the capacity of MSCs to differentiate into different cells assessed by gene expression. Results of this experiments will explain whether a gene plays a role in non-bone related biological processes. (B) Next, in addition to cell culture experiments, we will examine the physiological role of target genes by employing zebrafish animal model in the collaboration with Bar Ilan University. (C) Moreover, to elucidate the function of genetic targets in pathological conditions, correlation of target gene expression in patient tissue samples will be examined in collaboration with University Medical Centre and Orthopaedic Hospital Valdoltra. For that purpose bone, muscle and adipose tissues will be collected and gene expression analysed. Results of clinical part of the project will unveil the relevance of genetic targets in pathological processes in humans and provide crucial information on medical relevance of examined targets and determine the predictive potential of the genetic loci. (D) Finally, in the collaboration with National Institute of Chemistry, we will analyze whether genetic loci form a specific three-dimensional DNA structure that may explain alteration of their function. The proposed project is one of the first comprehensive studies that will functionally evaluate several osteoporosis associated genetic loci in vitro, in vivo and in clinical studies. Moreover, the role of genetic targets will be simultaneously examined in three different cell types (bone, fat, muscle) and in animal model. Therefore, we expect to obtain important results that will unveil the relevance of potential biomarkers/drug targets from different perspectives and thus significantly contribute also to the field of bone biology research. In summary, we will develop a pipeline for identification of novel drug targets for anabolic treatment of osteoporosis and provide fundamental advances in the field of bone biology. Importantly, our study may lead to discovery of novel biomarkers of fracture risks assessments with the direct implications in clinical practice.

In the proposed project, GWAS hits will be functionally characterised from different perspectives, leading to comprehensive and relevant results required for decision making process for choosing appropriate drug targets for novel treatments of osteoporosis. The major result of the project will be a set of very well characterised gene targets suitable for drug development and personalized based medicine for prevention of osteoporosis. Importantly, we will set up the methods that will serve as a system for functional characterization of any other candidate gene that will be identified as potential target interfering with bone remodelling, fat or muscle metabolism. Given the comprehensive approach, we expect to provide fundamental advances in the field of bone biology. Besides strengthening international collaborations with outstanding researches from abroad, and connecting basic and clinical research in Slovenia, this project will also be part of education process at the Faculty of Pharmacy. Two PhD students (already enrolled in the PhD Program) and several master students will be recruited to work on the project. Notably, this project is part of an ongoing collaboration within international GEFOS consortium (financed by EU COST project GEMSTONE), which has already led to outstanding scientific findings. Given that current project involves international collaboration with excellent foreign scientist and the topic is extremely hot this promises highly relevant results of the proposed study.

In the proposed project, GWAS hits will be functionally characterised from different perspectives, leading to comprehensive and relevant results required for decision making process for choosing appropriate drug targets for novel treatments of osteoporosis. The major result of the project will be a set of very well characterised gene targets suitable for drug development and personalized based medicine for prevention of osteoporosis. Importantly, we will set up the methods that will serve as a system for functional characterization of any other candidate gene that will be identified as potential target interfering with bone remodelling, fat or muscle metabolism. Given the comprehensive approach, we expect to provide fundamental advances in the field of bone biology. Besides strengthening international collaborations with outstanding researches from abroad, and connecting basic and clinical research in Slovenia, this project will also be part of education process at the Faculty of Pharmacy. Two PhD students (already enrolled in the PhD Program) and several master students will be recruited to work on the project. Notably, this project is part of an ongoing collaboration within international GEFOS consortium (financed by EU COST project GEMSTONE), which has already led to outstanding scientific findings. Given that current project involves international collaboration with excellent foreign scientist and the topic is extremely hot this promises highly relevant results of the proposed study.