The primary goals of the work proposed here are: 1) to examine the evolutionary history of the Type X collagen gene and to establish its relationship to other genes encoding proteins with collagenous domains, 2) to examine the biosynthesis of the molecule, and its potential role in the physiological removal of cartilage during endochondral ossification, 3) to examine the expression of Types II and X collagens during embryonic development and maturation of cartilage, and 4) to determine the molecular mechanisms of the high level of expression of Type X collagen in hypertrophic cartilage. To accomplish these goals, we will use a combination of DNA cloning/sequencing and protein chemistry. Type X collagen genes will be isolated from rat and human genomic libraries and their structure, as determined by nucleotide sequence analysis, will be compared to that of the chicken gene which we have isolated. Nucleotide sequences for chicken, rat, and human Type X collagens will be used to synthesize peptides. These will be used as haptens to generate specific antibodies. Such antibodies, together with specific DNA probes, will be used to study the synthesis of Type X collagen and the expression of the Type X gene during normal and abnormal cartilage development. Isolated cDNA and genomic clones will allow studies of the molecular mechanisms responsible for the expression of Type X collagen by hypertrophic chondrocytes. Nuclease hypersensitivity and methylation in and around the Type X gene will be mapped in expressing and non-expressing chondrocytes. The transient expression of the cloned genes will be studied in cultured cells, and we will assay for the binding of nuclear proteins to sequences within and flanking the gene. The long term objective of these studies is to understand the biogenesis and cellular regulation of extracellular matrices during normal and abnormal development. The studies on Type X collagen should provide information about the function and mechanisms controlling the synthesis of a specific collagen in a connective tissue undergoing morphogenetic destruction. They may also provide more general insight into the synthesis of extracellular matrix components by chondrocytes in normal cartilage and in disease states such as rheumatoid arthritis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR036819-06
Application #
3157744
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1985-07-01
Project End
1991-12-31
Budget Start
1990-01-01
Budget End
1990-12-31
Support Year
6
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Hu, Kai; Olsen, Bjorn R; Besschetnova, Tatiana Y (2017) Cell autonomous ANTXR1-mediated regulation of extracellular matrix components in primary fibroblasts. Matrix Biol 62:105-114
Nagao, Masashi; Hamilton, John L; Kc, Ranjan et al. (2017) Vascular Endothelial Growth Factor in Cartilage Development and Osteoarthritis. Sci Rep 7:13027
Hu, Kai; Olsen, Bjorn R (2017) Vascular endothelial growth factor control mechanisms in skeletal growth and repair. Dev Dyn 246:227-234
Hu, Kai; Besschetnova, Tatiana Y; Olsen, Bjorn R (2017) Soluble VEGFR1 reverses BMP2 inhibition of intramembranous ossification during healing of cortical bone defects. J Orthop Res 35:1461-1469
Hu, Kai; Olsen, Bjorn R (2016) Osteoblast-derived VEGF regulates osteoblast differentiation and bone formation during bone repair. J Clin Invest 126:509-26
Olsen, Bjorn R; Berendsen, Agnes D; Besschetnova, Tatiana Y et al. (2016) Fell-Muir Lecture: Regulatory mechanisms of skeletal and connective tissue development and homeostasis - lessons from studies of human disorders. Int J Exp Pathol 97:296-302
Hu, Kai; Olsen, Bjorn R (2016) The roles of vascular endothelial growth factor in bone repair and regeneration. Bone 91:30-8
Huang, Wei; Li, Qing; Amiry-Moghaddam, Mahmood et al. (2016) Critical Endothelial Regulation by LRP5 during Retinal Vascular Development. PLoS One 11:e0152833
Duan, Xuchen; Bradbury, Seth R; Olsen, Bjorn R et al. (2016) VEGF stimulates intramembranous bone formation during craniofacial skeletal development. Matrix Biol 52-54:127-140
Nagao, M; Cheong, C W; Olsen, B R (2016) Col2-Cre and tamoxifen-inducible Col2-CreER target different cell populations in the knee joint. Osteoarthritis Cartilage 24:188-91

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