A regulatory cascade involving retinoic acid, Cbfa1, and matrix metalloproteinases is coupled to the development of a process of perichondrial invasion and osteogenic differentiation during bone formation

Abstract

Tissue-remodeling processes are largely mediated by members of the matrix metalloproteinase (MMP) family of endopeptidases whose expression is strictly controlled both spatially and temporally. In this article, we have examined the molecular mechanisms that could contribute to modulate the expression of MMPs like collagenase-3 and MT1-MMP during bone formation. We have found that all-trans retinoic acid (RA), which usually downregulates MMPs, strongly induces collagenase-3 expression in cultures of embryonic metatarsal cartilage rudiments and in chondrocytic cells. This effect is dose and time dependent, requires the de novo synthesis of proteins, and is mediated by RAR-RXR heterodimers. Analysis of the signal transduction mechanisms underlying the upregulating effect of RA on collagenase-3 expression demonstrated that this factor acts through a signaling pathway involving p38 mitogen-activated protein kinase. RA treatment of chondrocytic cells also induces the production of MT1-MMP, a membrane-bound metalloproteinase essential for skeletal formation, which participates in a proteolytic cascade with collagenase-3. The production of these MMPs is concomitant with the development of an RA-induced differentiation program characterized by formation of a mineralized bone matrix, downregulation of chondrocyte markers like type II collagen, and upregulation of osteoblastic markers such as osteocalcin. These effects are attenuated in metatarsal rudiments in which RA induces the invasion of perichondrial osteogenic cells from the perichondrium into the cartilage rudiment. RA treatment also resulted in the upregulation of Cbfa1, a transcription factor responsible for collagenase-3 and osteocalcin induction in osteoblastic cells. The dynamics of Cbfa1, MMPs, and osteocalcin expression is consistent with the fact that these genes could be part of a regulatory cascade initiated by RA and leading to the induction of Cbfa1, which in turn would upregulate the expression of some of their target genes like collagenase-3 and osteocalcin.

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

“…MMP-13 has a particular affinity for type II collagen and selectively enhances cleavage and denaturation of this type of collagen (43), and can also cleave aggrecan at specific sites (44). Interestingly, recent studies have also shown that retinoid-related signals strongly enhance the induction of MMP-13 in chondrocytic cells through the action of RAR-RXR heterodimers (45). In addition, reduced expression of MMP-13 is observed in cytokine-stimulated cells when NF-B is maintained in the cytoplasm by constitutive levels of I B (46).…”

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

“…Because the expression of MMP-13 is elevated in the arthritic joints (35,39), this proteinase is thought to be a key enzyme associated with osteoarthritis. Recent studies have shown that retinoid-related signals strongly enhance the induction of MMP-13 in chondrocytic cells through the action of retinoic acid receptor-retinoid X receptor heterodimers (41). In addition, it has been demonstrated that MMP-13 induction by inflammatory cytokines requires the activation of NFB (42).…”

Distinct Transglutaminase 2-independent and Transglutaminase 2-dependent Pathways Mediate Articular Chondrocyte Hypertrophy

“…To directly probe TG2 functions in chondrocyte differentiation and calcification we have taken advantage of the recent generation of TG2 knockout mice, which are phenotypically normal (23,24). To specifically assess for direct TG2 involvement in articular chondrocyte hypertrophy, we studied the effects on primary chondrocytes of CNP (25) and of the vitamin A-derived metabolite all-trans retinoic acid (ATRA) (26,27). CNP is expressed in developing long bones, and the stimulatory effects of CNP on chondrocyte proliferation and hypertrophy are essential for normal growth plate development (25).…”

“…CNP is expressed in developing long bones, and the stimulatory effects of CNP on chondrocyte proliferation and hypertrophy are essential for normal growth plate development (25). ATRA promotes maturation to hypertrophy and calcification by chick sternal chondrocytes in vitro (26) in a manner that requires the transcription factor Cbfa1 (27). In addition, hypervitaminosis A and toxicity of certain other retinoids is associated with accelerated terminal differentiation of endochondral chondrocytes and premature epiphyseal closure in vivo (28,29).…”