title: Emerging techniques for the discovery and validation of therapeutic targets for skeletal diseases.
abstract: Advances in genomics and proteomics have revolutionised the drug discovery process and target validation. Identification of novel therapeutic targets for chronic skeletal diseases is an extremely challenging process based on the difficulty of obtaining high-quality human diseased versus normal tissue samples. The quality of tissue and genomic information obtained from the sample is critical to identifying disease-related genes. Using a genomics-based approach, novel genes or genes with similar homology to existing genes can be identified from cDNA libraries generated from normal versus diseased tissue. High-quality cDNA libraries are prepared from uncontaminated homogeneous cell populations harvested from tissue sections of interest. Localised gene expression analysis and confirmation are obtained through in situ hybridisation or immunohistochemical studies. Cells overexpressing the recombinant protein are subsequently designed for primary cell-based high-throughput assays that are capable of screening large compound banks for potential hits. Afterwards, secondary functional assays are used to test promising compounds. The same overexpressing cells are used in the secondary assay to test protein activity and functionality as well as screen for small-molecule agonists or antagonists. Once a hit is generated, a structure-activity relationship of the compound is optimised for better oral bioavailability and pharmacokinetics allowing the compound to progress into development. Parallel efforts from proteomics, as well as genetics/transgenics, bioinformatics and combinatorial chemistry, and improvements in high-throughput automation technologies, allow the drug discovery process to meet the demands of the medicinal market. This review discusses and illustrates how different approaches are incorporated into the discovery and validation of novel targets and, consequently, the development of potentially therapeutic agents in the areas of osteoporosis and osteoarthritis. While current treatments exist in the form of hormone replacement therapy, antiresorptive and anabolic agents for osteoporosis, there are no disease-modifying therapies for the treatment of the most common human joint disease, osteoarthritis. A massive market potential for improved options with better safety and efficacy still remains. Therefore, the application of genomics and proteomics for both diseases should provide much needed novel therapeutic approaches to treating these major world health problems.
authors.affiliation: GlaxoSmithKline Pharmaceuticals, Department of Musculoskeletal Diseases, UW2109, 709 Swedeland Rd, King of Prussia, PA 19406, USA.
authors.forename: Christine H
authors.initials: CH
authors.lastname: Cho
authors.forename: Mark E
authors.initials: ME
authors.lastname: Nuttall
mesh_terms: D000818:Animals; D001847:Bone Diseases / Q000188:drug therapy*; D002352:Carrier Proteins; D020169:Caspases / Q000502:physiology; D056657:Cathepsin K; D002403:Cathepsins / Q000037:antagonists & inhibitors; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000502:physiology; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D051766:Early Growth Response Protein 1; D023281:Genomics; D006023:Glycoproteins / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins / Q000502:physiology; D008562:Membrane Glycoproteins; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D015394:Molecular Structure; D010003:Osteoarthritis / Q000188:drug therapy / Q000378:metabolism; D010024:Osteoporosis / Q000188:drug therapy / Q000378:metabolism; D053244:Osteoprotegerin; D040901:Proteomics; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear / Q000187:drug effects / Q000502:physiology; D018124:Receptors, Tumor Necrosis Factor / Q000502:physiology; D014157:Transcription Factors / Q000502:physiology; D020798:Two-Hybrid System Techniques; D019061:src-Family Kinases
chemical_list: D002352:Carrier Proteins; D015853:Cysteine Proteinase Inhibitors; D004268:DNA-Binding Proteins; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D006023:Glycoproteins; D017874:Immediate-Early Proteins; D008562:Membrane Glycoproteins; D053244:Osteoprotegerin; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C505686:TNFRSF11A protein, human; C506030:TNFRSF11B protein, human; C505688:TNFSF11 protein, human; C505687:Tnfrsf11a protein, mouse; C506031:Tnfrsf11b protein, mouse; C505689:Tnfsf11 protein, mouse; D014157:Transcription Factors; D019061:src-Family Kinases; D002403:Cathepsins; D020169:Caspases; C534130:CTSK protein, human; D056657:Cathepsin K; C534132:Ctsk protein, mouse