Development of comprehensive functional genomic screens to identify novel mediators of osteoarthritis

Daouti, Sherif; Latario, Brian; Nagulapalli, Sujatha; Buxton, Frank P.; Uziel-Fusi, Susan; Chirn, Gung-Wei; Bodian, Dale L.; Song, Chi; Labow, Mark; Lotz, Martin; Quintavalla, Joseph; Kumar, C. Santhosh

doi:10.1016/j.joca.2005.02.003

Cited by 44 publications

(26 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2,26 Previous studies evaluated the expression levels of collagenases, cytokines and growth factors in experimental animal models of OA, which mimic the disease in humans. 27,28 However, those studies were primarily restricted to examine molecular changes in the cartilage layer without integrating information from the surrounding synovial membrane or subchondral bone in response to the disease. In more recent studies, researchers have studied the crosstalk between articular cartilage and synovial membrane.…”

Section: Discussionmentioning

confidence: 99%

The effects of interleukin-1β in modulating osteoclast-conditioned medium’s influence on gelatinases in chondrocytes through mitogen-activated protein kinases

Xie

Cai

et al. 2015

Int J Oral Sci

View full text Add to dashboard Cite

Osteoarthritis is recognised to be an interactive pathological process involving the cartilage, subchondral bone and synovium. The signals from the synovium play an important role in cartilage metabolism, but little is known regarding the influence of the signalling from bone. Additionally, the collagenases and stromelysin-1 are involved in cartilage catabolism through mitogen-activated protein kinase (MAPK) signalling, but the role of the gelatinases has not been elucidated. Here, we studied the influence of osteoclastic signals on chondrocytes by characterising the expression of interleukin-1β (IL-1β)-induced gelatinases through MAPK signalling. We found that osteoclast-conditioned media attenuated the gelatinase activity in chondrocytes. However, IL-1β induced increased levels of gelatinase activity in the conditioned media group relative to the mono-cultured chondrocyte group. More specifically, IL-1β restored high levels of gelatinase activity in c-Jun N-terminal kinase inhibitor-pretreated chondrocytes in the conditioned media group and led to lower levels of gelatinase activity in extracellular signal-regulated kinase or p38 inhibitor-pretreated chondrocytes. Gene expression generally correlated with protein expression. Taken together, these results show for the first time that signals from osteoclasts can influence gelatinase activity in chondrocytes. Furthermore, these data show that IL-1β restores gelatinase activity through MAPK inhibitors; this information can help to increase the understanding of the gelatinase modulation in articular cartilage.

show abstract

Section: Discussionmentioning

confidence: 99%

The effects of interleukin-1β in modulating osteoclast-conditioned medium’s influence on gelatinases in chondrocytes through mitogen-activated protein kinases

Xie

Cai

et al. 2015

Int J Oral Sci

View full text Add to dashboard Cite

show abstract

“…37,39,40 Recently, Axl polymorphisms were associated with salt-induced hypertension 38 and osteoarthritis. 41 An important role for the Gas6/Axl/Akt pathway in thrombosis has been shown because both Gas6 and Axlfamily receptor knockout mice exhibited abnormalities in platelet function, 13,14 and a Gas6 mutation is associated with thrombotic stroke in humans. 12 However, it is likely that Gas6-dependent signaling is important only during pathological processes because no correlation between plasma Gas6 and platelet aggregation in healthy people was found.…”

Section: Discussionmentioning

confidence: 99%

Axl, A Receptor Tyrosine Kinase, Mediates Flow-Induced Vascular Remodeling

Korshunov

Mohan

Georger

et al. 2006

Circulation Research

102

View full text Add to dashboard Cite

Abstract-Intima-media thickening (IMT) in response to hemodynamic stress is a physiological process that requires coordinated signaling among endothelial, inflammatory, and vascular smooth muscle cells (VSMC). Axl, a receptor tyrosine kinase, whose ligand is Gas6, is highly induced in VSMC after carotid injury. Because Axl regulates cell migration, phagocytosis and apoptosis, we hypothesized that Axl would play a role in IMT. Vascular remodeling in mice deficient in Axl (Axl Ϫ/Ϫ ) and wild-type littermates (Axl ϩ/ϩ ) was induced by ligation of the left carotid artery (LCA) branches maintaining flow via the left occipital artery. Both genotypes had similar baseline hemodynamic parameters and carotid artery structure. Partial ligation altered blood flow equally in both genotypes: increased by 60% in the right carotid artery (RCA) and decreased by 80% in the LCA. There were no significant differences in RCA remodeling between genotypes. However, in the LCA Axl Ϫ/Ϫ developed significantly smaller intimaϩmedia compared with Axl Key Words: Axl Ⅲ flow Ⅲ carotid artery Ⅲ remodeling Ⅲ mouse Ⅲ apoptosis Ⅲ inflammation T he receptor tyrosine kinase Axl (also known as Ufo and Tyro7) belongs to a family of tyrosine receptors that includes Tyro3 (Sky) and Mer (Tyro12). 1,2 A common ligand for Axl family is Gas6 (Growth arrest-specific protein 6). 2,3 Important cellular functions of Gas6/Axl include cell adhesion, migration, phagocytosis, and inhibition of apoptosis. 4 -7 Gas6 and Axl family receptors are highly regulated in a tissue and disease specific manner. 8 -11 A recent genetic study found a significant association of a Gas6 mutation with stroke in humans. 12 Both Gas6 and Axl-family receptor knockout mice show abnormalities in platelet function, further supporting a vascular role for the Gas6/Axl pathway. 13,14 In human umbilical vein endothelial cells (HUVEC) and pulmonary artery EC, Gas6/Axl signaling promotes cell survival, 8,15 possibly via an autocrine pathway. 16 Gas6/Axlmediated survival signals include activation of Akt, phosphorylation of nuclear factor B (NF-B), increased expression of Bcl-2, and decreased caspase-3 activation. 17 Similarly, in vascular smooth muscle cells (VSMC), Gas6/Axl powerfully stimulates cell migration and cell survival. 18 We reported that Axl was highly regulated after rat carotid balloon injury consistent with a role in intimal VSMC proliferation. 19 Our recent findings showed that the Gas6/Axl/PI3K/Akt pathway inhibited VSMC apoptosis. 20 Moreover, Axl contributes to generation of reactive oxygen species in VSMC. 21 In addition, the Gas6/Axl pathway is important in the mammalian immune system. 7 Recent data suggest that the Axl family is a central regulator of macrophage activation and phagocytosis. 22 We developed a reproducible mouse model of flowdependent vascular remodeling that resembles human intimamedia thickening (IMT). 23 In response to decreased blood flow IMT occurs, which involves critical interactions among cells of the vessel wall. 23,24 Based on the significant rol...

show abstract

“…of 5-10) harboring the SV40 large T antigen without any drug resistance or selectable markers. After 48 h postinfection, cells from each donor were detached with trypsin (Sigma, St. Louis, MS) treatment, pooled and divided into three parts: (1) one part was replated as monolayer in multiple 100 mm culture dishes for clone selection; (2) one part was encapsulated in 0.4% low melting agarose and layered over 0.7% agarose and cultured for 4 weeks according to a previously reported procedure (Daouti et al, 2005); and (3) one part was encapsulated in 1.2% alginate beads and cultured for 4 weeks according to a previously reported procedure (Majumdar et al, 2000).…”

Section: Chondrocyte Immortalization and Clone Expansionmentioning

confidence: 99%

Development and characterization of a human articular cartilage‐derived chondrocyte cell line that retains chondrocyte phenotype

Hoffman

Newman‐Tarr

Gibbard

et al. 2009

Journal Cellular Physiology

View full text Add to dashboard Cite

Chondrocytes, the only cell type present in articular cartilage, regulate tissue homeostasis by a fine balance of metabolism that includes both anabolic and catabolic activities. Therefore, the biology of chondrocytes is critical for understanding cartilage metabolism. One major limitation when studying primary chondrocytes in culture is their loss of phenotype. To overcome this hurdle, limited attempts have been made to develop human chondrocyte cell lines that retain the phenotype for use as a good surrogate model. In this study, we report a novel approach to the establishment and characterization of human articular cartilage-derived chondrocyte cell lines. Adenoviral infection followed by culture of chondrocytes in 3-dimensional matrix within 48 h post-infection maintained the phenotype prior to clonal selection. Cells were then placed in culture either as monolayer, or in 3-dimensional matrix of alginate or agarose. The clones were characterized by their basal gene expression profile of chondrocyte markers. Based on type II collagen expression, 21 clones were analyzed for gene expression following treatment with IL-1 or BMP-7 and compared to similarly stimulated primary chondrocytes. This resulted in selection of two clones that retained the chondrocyte phenotype as evidenced by expression of type II collagen and other extra-cellular matrix molecules. In addition, one clone (AL-4-17) showed similar responses as primary chondrocytes when treated with IL-1 or BMP-7. In summary, this report provides a novel procedure to develop human articular cartilage-derived chondrocyte cell lines, which preserve important characteristics of articular chondrocytes and represent a useful model to study chondrocyte biology.

show abstract

Development of comprehensive functional genomic screens to identify novel mediators of osteoarthritis

Cited by 44 publications

References 38 publications

The effects of interleukin-1β in modulating osteoclast-conditioned medium’s influence on gelatinases in chondrocytes through mitogen-activated protein kinases

The effects of interleukin-1β in modulating osteoclast-conditioned medium’s influence on gelatinases in chondrocytes through mitogen-activated protein kinases

Axl, A Receptor Tyrosine Kinase, Mediates Flow-Induced Vascular Remodeling

Development and characterization of a human articular cartilage‐derived chondrocyte cell line that retains chondrocyte phenotype

Contact Info

Product

Resources

About