Skeletogenesis proceeds through a series of phases in which resting chondrocytes within a cartilaginous anlage initiate proliferation, enter a hypertrophic state, and are replaced by osteoblasts that mineralize the surrounding matrix. This process is controlled by hormones and local effector molecules. In vitro the transforming growth factor betas (TGF- betas) have profound effects on both chondrocytes and osteoblasts. In vivo TGF-beta is an inhibitor of articular chondroctye differentiation, whereas in bone, TGF-beta overexpression causes a regional differentiation of osteoblasts to osteocytes and an osteoporotic state. These somewhat limited effects contrast with the widespread distribution of the TGF-betas in cartilage and bone as well as multiple skeletal defects in TGF-beta2 null mouse. We hypothesize that the involvement of TGF-beta in skeletogenesis is more extensive than reported because previous approaches failed to consider that TGF-beta action is controlled at the level of conversion of extracellular latent complex to the active form. We will examine the role of TGF-beta in cartilage and bone by using genetically manipulated mice that express either increased or decreased levels of active TGF-beta. We will manipulate the conversion of the normally latent TGF-beta complex, consisting of TGF-beta, the TGF-beta propeptide, and the latent TGF- beta binding protein (LTBP), to its active form, rather than altering TGF- beta gene expression or signaling. First, we will generate transgenic mice that express a form of LTBP that enhances activation of latent TGF-beta. Second, we will produce transgenic mice expressing a form of LTBP that suppresses activation. Each construct will be expressed using the collagen I promoter to target effects to early osteoblasts, the collagen II promoter to see effects during chondrocyte differentiation, and the osteocalcin promoter to establish the effects on osteoblast physiology. In addition; we will create a mouse in which the cysteine residue required for TGF-beta2 bonding to LTBP is mutated to serine precluding interaction of TGF-beta with LTBP resulting in excess TGF-beta activation. In vivo and in vitro histological and biochemical analysis of these animals should reveal phenotypes that yield insight into the role of TGF-beta in chondrogenesis and osteoblast biology. These experiments will contribute to the overall IRPG on Skeletal Modeling and Remodeling through interactions with other investigators interested in morphogenesis-(Project 1) and signaling (Project 2).

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
3R01DE013742-01A1S1
Application #
6500629
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Program Officer
Zhang, Guo He
Project Start
2001-02-01
Project End
2004-12-31
Budget Start
2001-09-01
Budget End
2001-12-31
Support Year
1
Fiscal Year
2001
Total Cost
$10,431
Indirect Cost
Name
New York University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10016
Dabovic, B; Levasseur, R; Zambuto, L et al. (2005) Osteopetrosis-like phenotype in latent TGF-beta binding protein 3 deficient mice. Bone 37:25-31
Ramirez, Francesco; Sakai, Lynn Y; Dietz, Harry C et al. (2004) Fibrillin microfibrils: multipurpose extracellular networks in organismal physiology. Physiol Genomics 19:151-4
Annes, Justin P; Chen, Yan; Munger, John S et al. (2004) Integrin alphaVbeta6-mediated activation of latent TGF-beta requires the latent TGF-beta binding protein-1. J Cell Biol 165:723-34
Annes, Justin P; Munger, John S; Rifkin, Daniel B (2003) Making sense of latent TGFbeta activation. J Cell Sci 116:217-24
Lack, Jeremy; O'Leary, Joanne M; Knott, Vroni et al. (2003) Solution structure of the third TB domain from LTBP1 provides insight into assembly of the large latent complex that sequesters latent TGF-beta. J Mol Biol 334:281-91
Ramirez, Francesco; Rifkin, Daniel B (2003) Cell signaling events: a view from the matrix. Matrix Biol 22:101-7
Isogai, Zenzo; Ono, Robert N; Ushiro, Shin et al. (2003) Latent transforming growth factor beta-binding protein 1 interacts with fibrillin and is a microfibril-associated protein. J Biol Chem 278:2750-7
Dabovic, Branka; Chen, Yan; Colarossi, Cristina et al. (2002) Bone abnormalities in latent TGF-[beta] binding protein (Ltbp)-3-null mice indicate a role for Ltbp-3 in modulating TGF-[beta] bioavailability. J Cell Biol 156:227-32
Dabovic, B; Chen, Y; Colarossi, C et al. (2002) Bone defects in latent TGF-beta binding protein (Ltbp)-3 null mice; a role for Ltbp in TGF-beta presentation. J Endocrinol 175:129-41