Proteases and their Involvement in Health and Disease
Our research is focused on the physiological and pathological role of cysteine proteases in extracellular matrix degradation and their functions in regulatory pathways. This includes the following mix of basic and translational research avenues and a wide spectrum of experimental methods (molecular biology, enzymology/biochemistry, crystallography and rational drug design, screening methods, cell culture techniques, and animal models including pharmacokinetic evaluation). The research strategies comprise:
• Drug target identification and characterization using molecular cloning and expression, structural analysis, and classical biochemistry approaches;
• Identification of novel inhibitors and their inhibitory mechanisms;
• Drug target validation using various enzymatic, cell-based, and murine models (transgenics and knockouts). Major disease and clinical indications include: osteoporosis, arthritis, atherosclerosis, jaw erosion, dentin-bounding stability, and skin ageing.
• Graduate Research Seminars (Faculty of Dentistry)
• Co-Chair in Biochem 551(Advanced topics in Biochemistry and Molecular Biology, Department of BMB)
• Graduate Student and Post-Doctoral Fellow supervision
!! Research Opportunities !!
MSc/PhD Graduate Student and Post-Doctoral Fellow positions in crystallography and computational modeling, proteomics, cellular biology, and murine disease models are available (For details see job postings.).
Dr. Dieter Brömme received his formal training (MSc and PhD) in Biochemistry from the Martin-Luther Universität (MLU) Halle-Wittenberg in Germany. The Halle-Wittenberg University was founded in 1502 (Wittenberg) and in 1694 (Halle) and represented the first modern research-driven university in Europe after which most European, North-American, and Japanese universities have been modeled. After graduation in the field of microbial proteases, Dr. Brömme worked as an academic scientist at the Institute of Biochemistry at the Faculty of Medicine at MLU and discovered his life-long passion for mammalian lysosomal cysteine proteases. Prior to joining the University of British Columbia in Vancouver, BC as a Tier I Canada Research Chair in 2004, he was a Senior Research Officer at the Biotechnology Research Institute in Montreal, QC (1991-1993), a project leader at the Biotech company Khepri Pharmaceuticals in San Francisco, CA (1993-1996), and an Associate/Full Professor at the Mount Sinai School of Medicine in New York, NY (1996-2004).
Dr. Brömme has 209 entries in the Web of Sciences with an H-index of 60 (2018). Currently, there are 180 peer-reviewed publications listed on PubMed, which include publications in Structural Biology, PNAS, J Clin Investigations, and various leading Research Society and specialty journals in cell biology, biochemistry, skeletal, and cardiovascular research. His laboratory attracts collaborations, as well as trainees (summer students, graduate students, post-doctoral fellows, and visiting scientists), from all continents and has been continuously funded by federal and society-based US and Canadian Institutions such as the NIH, CIHR, and NSERC.
For full bibliography see: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=search&DB=pubmed
Research in the Brömme Laboratory: Cathepsins and their Role in Health and Disease
Many human diseases are characterized by an excessive proteolytic degradation of proteins of the extracellular matrix (e.g., bone and cartilage diseases, atherosclerosis, lung diseases, cancer) or by inappropriate proteolytic processing of proteins – leading to auto-immune diseases and disorders caused by regulatory defects. Our laboratory is at the cutting edge of research into the identification of novel therapeutic targets among intracellular lysosomal proteases and is focused on the role of these proteases in the pathogenesis of osteoporosis, rheumatoid arthritis, atherosclerosis, dental diseases, skin aging, and certain forms of immune disorders. Our aim is to understand the role of lysosomal proteases in health and disease, which will lead to new therapeutic approaches to treat these illnesses. To achieve our objective, we exploit an interdisciplinary approach which includes methods of molecular biology, enzymology, crystallography, cell biology, histology, animal models, and various high-resolution imaging techniques.
Our laboratory was at the forefront in the identification and characterization of several human cysteine cathepsins (Cathepsins S, K (O2), V, F, and W (1-5)). We solved the first Cathepsin K structure (6), along with its collagenolytically active complexes containing glycosaminoglycans and the CatK-odanacatib structure (7-10). We also developed the concept of Affinity Crystallography to simultaneously identify and solve the structure of novel protease inhibitors from crude natural extracts (11).
Some cathepsins have been implicated in the regulation of antigen processing (Cathepsins S, V, and F (12,13)) and extracellular matrix degradation (Cathepsins K, V, and S (2,14-18)). Here we have primarily focused on the degradation of the two most common ECM proteins: collagen and elastin. We demonstrated early on that CatK is a unique and powerful collagenase predominantly expressed in osteoclasts (19-21), which made this protease a long-sought-after drug target for the treatment of skeletal disorders, such as osteoporosis and arthritis (14,22), and cardiovascular diseases (23,24). Although highly potent and selective inhibitors were developed by the pharmaceutical industry and proven to be efficacious in clinical trials, none were approved due to the onslaught of cardiovascular and skin side effects (25). Thus in recent years, we directed our efforts toward understanding these side effects and have developed the concept of “Substrate-specific Ectosteric Inhibition of proteases” which allows us to target the disease-relevant activity of a target enzyme without affecting its multiple other functions (26). The inhibition of regulatory functions of the ECM proteases by so-called active site-directed inhibitors leads to the observed side effects. We are currently identifying and optimizing new ectosteric inhibitors of cathepsins by means of various high-throughput enzymatic and computational screening approaches (27). Interestingly, Chinese medicinal herbs are a rich source of ectosteric inhibitors (28,29). We are applying this concept on diseases where extracellular matrix degradation is the major disease-defining event. These include skeletal and dental diseases such as osteoporosis (29,30), arthritis, bone cancer, jaw bone erosion, vascular diseases, and skin ageing (31). Our initial attempt to apply the concept of ectosteric inhibitors in an osteoporosis mouse model received wide-spread international media coverage, as it promises efficacy and less side effects than current therapies. (https://www.genengnews.com/gen-news-highlights/traditional-chinese-medicine-opens-new-path-to-treatment-of-bone-loss/81254864; https://www.hindustantimes.com/health/bone-cure-this-chinese-herb-may-treat-people-with-osteoporosis/story-mDXX6dNJlRE9cw6mhXRqMJ.html; https://www.pharmaceutical-technology.com/features/red-sage-ancient-chinese-herb-prevent-bone-loss/
Potential trainees and collaborators who are interested in our work are advised to explore previous and current publications listed in PubMed (https://www.ncbi.nlm.nih.gov/pubmed/?term=Bromme-d and https://www.ncbi.nlm.nih.gov/pubmed/?term=Broemme-d) to get more insight into the research of the Brömme laboratory.
1. Bromme, D., Bonneau, P. R., Lachance, P., Wiederanders, B., Kirschke, H., Peters, C., Thomas, D. Y., Storer, A. C., and Vernet, T. (1993) Functional expression of human cathepsin S in Saccharomyces cerevisiae. Purification and characterization of the recombinant enzyme. J Biol Chem 268, 4832-4838
2. Brömme, D., Okamoto, K., Wang, B. B., and Biroc, S. (1996) Human cathepsin O2, a matrix protein-degrading cysteine protease expressed in osteoclasts. Functional expression of human cathepsin O2 in Spodoptera frugiperda and characterization of the enzyme. J Biol Chem 271, 2126-2132
3. Brömme, D., Li, Z., Barnes, M., and Mehler, E. (1999) Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry 38, 2377-2385
4. Wang, B., Shi, G. P., Yao, P. M., Li, Z., Chapman, H. A., and Bromme, D. (1998) Human cathepsin F. J Biol Chem 273, 32000-32008
5. Linnevers, C., Smeekens, S. P., and Bromme, D. (1997) Human cathepsin W, a putative cysteine protease predominantly expressed in CD8+ T-lymphocytes. FEBS Lett 405, 253-259
6. McGrath, M. E., Klaus, J. L., Barnes, M. G., and Bromme, D. (1997) Crystal structure of human cathepsin K complexed with a potent inhibitor. Nat Struct Biol 4, 105-109.
7. Li, Z., Hou, W. S., Escalante-Torres, C. R., Gelb, B. D., and Bromme, D. (2002) Collagenase activity of cathepsin K depends on complex formation with chondroitin sulfate. J Biol Chem 277, 28669-28676
8. Li, Z., Kienetz, M., Cherney, M. M., James, M. N., and Bromme, D. (2008) The crystal and molecular structures of a cathepsin K:chondroitin sulfate complex. J Mol Biol 383, 78-91
9. Cherney, M. M., Lecaille, F., Kienitz, M., Nallaseth, F. S., Li, Z., James, M. N., and Bromme, D. (2011) Structure-activity analysis of cathepsin K/chondroitin 4-sulfate interactions. J Biol Chem 286, 8988-8998
10. Aguda, A. H., Panwar, P., Du, X., Nguyen, N. T., Brayer, G. D., and Bromme, D. (2014) Structural basis of collagen fiber degradation by cathepsin K. Proc Natl Acad Sci U S A 111, 17474-17479
11. Aguda, A. H., Lavallee, V., Cheng, P., Bott, T. M., Meimetis, L. G., Law, S., Nguyen, N. T., Williams, D. E., Kaleta, J., Villanueva, I., Davies, J., Andersen, R. J., Brayer, G. D., and Bromme, D. (2016) Affinity Crystallography: A New Approach to Extracting High-Affinity Enzyme Inhibitors from Natural Extracts. J Nat Prod 79, 1962-1970
12. Shi, G. P., Bryant, R. A., Riese, R., Verhelst, S., Driessen, C., Li, Z., Bromme, D., Ploegh, H. L., and Chapman, H. A. (2000) Role for cathepsin F in invariant chain processing and major histocompatibility complex class II peptide loading by macrophages. J Exp Med 191, 1177-1186
13. Tolosa, E., Li, W., Yasuda, Y., Wienhold, W., Denzin, L. K., Lautwein, A., Driessen, C., Schnorrer, P., Weber, E., Stevanovic, S., Kurek, R., Melms, A., and Bromme, D. (2003) Cathepsin V is involved in the degradation of invariant chain in human thymus and is overexpressed in myasthenia gravis. J Clin Invest 112, 517-526
14. Hou, W. S., Li, Z., Gordon, R. E., Chan, K., Klein, M. J., Levy, R., Keysser, M., Keyszer, G., and Bromme, D. (2001) Cathepsin K is a critical protease in synovial fibroblast-mediated collagen degradation. Am J Pathol 159, 2167-2177.
15. Hou, W. S., Li, Z., Buttner, F. H., Bartnik, E., and Bromme, D. (2003) Cleavage site specificity of cathepsin K toward cartilage proteoglycans and protease complex formation. Biol Chem 384, 891-897
16. Yasuda, Y., Li, Z., Greenbaum, D., Bogyo, M., Weber, E., and Bromme, D. (2004) Cathepsin V, a novel and potent elastolytic activity expressed in activated macrophages. J Biol Chem 279, 36761-36770
17. Panwar, P., Du, X., Sharma, V., Lamour, G., Castro, M., Li, H., and Bromme, D. (2013) Effects of cysteine proteases on the structural and mechanical properties of collagen fibers. J Biol Chem 288, 5940-5950
18. Panwar, P., Lamour, G., Mackenzie, N. C., Yang, H., Ko, F., Li, H., and Bromme, D. (2015) Changes in Structural-Mechanical Properties and Degradability of Collagen during Aging-associated Modifications. J Biol Chem 290, 23291-23306
19. Bromme, D., and Okamoto, K. (1995) Human cathepsin O2, a novel cysteine protease highly expressed in osteoclastomas and ovary molecular cloning, sequencing and tissue distribution. Biol Chem Hoppe Seyler 376, 379-384.
20. Kafienah, W., Bromme, D., Buttle, D. J., Croucher, L. J., and Hollander, A. P. (1998) Human cathepsin K cleaves native type I and II collagens at the N-terminal end of the triple helix. Biochem J 331, 727-732
21. Xia, L., Kilb, J., Wex, H., Lipyansky, A., Breuil, V., Stein, L., Palmer, J. T., Dempster, D. W., and Brömme, D. (1999) Localization of rat cathepsin K in osteoclasts and resorption pits: Inhibition of bone resorption cathepsin K-activity by peptidyl vinyl sulfones. Biol Chem 380, 679-687
22. Hou, W.-S., Li, W., Keyszer, G., Weber, E., Levy, R., Klein, M. J., Gravallese, E. M., Goldring, S. R., and Bromme, D. (2002) Comparison of cathepsins K and S expression within the rheumatoid and osteoarthritic synovium. Arthritis Rheum 46, 663-674
23. Samokhin, A. O., Buhling, F., Theissig, F., and Bromme, D. (2010) ApoE-deficient mice on cholate-containing high-fat diet reveal a pathology similar to lung sarcoidosis. Am J Pathol 176, 1148-1156
24. Samokhin, A. O., Lythgo, P. A., Gauthier, J. Y., Percival, M. D., and Bromme, D. (2010) Pharmacological inhibition of cathepsin S decreases atherosclerotic lesions in Apoe-/- mice. J Cardiovasc Pharmacol 56, 98-105
25. Bromme, D., Panwar, P., and Turan, S. (2016) Cathepsin K osteoporosis trials, pycnodysostosis and mouse deficiency models: Commonalities and differences. Expert Opin Drug Discov 11, 457-472
26. Panwar, P., Soe, K., Guido, R. V., Bueno, R. V., Delaisse, J. M., and Bromme, D. (2016) A novel approach to inhibit bone resorption: exosite inhibitors against cathepsin K. Br J Pharmacol 173, 396-410
27. Law, S., Andrault, P. M., Aguda, A. H., Nguyen, N. T., Kruglyak, N., Brayer, G. D., and Bromme, D. (2017) Identification of mouse cathepsin K structural elements that regulate the potency of odanacatib. Biochem J 474, 851-864
28. Guo, Y., Li, Y., Xue, L., Severino, R. P., Gao, S., Niu, J., Qin, L. P., Zhang, D., and Bromme, D. (2014) Salvia miltiorrhiza: an ancient Chinese herbal medicine as a source for anti-osteoporotic drugs. J Ethnopharmacol 155, 1401-1416
29. Panwar, P., Law, S., Jamroz, A., Azizi, P., Zhang, D. W., Ciufolini, M., and Bromme, D. (2018) Tanshinones that selectively block the collagenase activity of cathepsin K provide a novel class of ectosteric antiresorptive agents for bone. Brit J Pharmacol 175, 902-923
30. Panwar, P., Xue, L., Soe, K., Srivastava, K., Law, S., Delaisse, J. M., and Bromme, D. (2017) An Ectosteric Inhibitor of Cathepsin K Inhibits Bone Resorption in Ovariectomized Mice. J Bone Miner Res 32, 2415-30
31. Panwar, P., Butler, G. S., Jamroz, A., Azizi, P., Overall, C. M., and Bromme, D. (2018) Aging-associated modifications of collagen affect its degradation by matrix metalloproteinases. Matrix Biol 65, 30-44