Transforming growth factor-β1 regulates fibronectin isoform expression and splicing factor SRp40 expression during ATDC5 chondrogenic maturation

Han, Fei; Gilbert, James R.; Harrison, Greg J.; Adams, Christopher S.; Freeman, Theresa A.; Tao, Zhuliang; Zaka, Raihana; Liang, Hongyan; Williams, Charlene J.; Tuan, Rocky S.; Norton, Pamela A.; Hickok, Noreen J.

doi:10.1016/j.yexcr.2007.01.028

Cited by 31 publications

(28 citation statements)

References 43 publications

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“…In light of the above data implicating ECM stiffness and cell contractility as modulators of EDB-FN splicing, and because SRp40 is one of the main regulators of EDB-FN expression (11,12), we extended our analysis to the SR proteins. Remarkably, phosphorylation levels of SR proteins increased with matrix stiffness in vitro (Fig.…”

Section: Ecm Stiffness Regulates Srp40 Phosphorylation In Culture and Inmentioning

confidence: 99%

“…EDB-FN is produced by endothelial cells (ECs) in tumors and may favor angiogenesis (7)(8)(9). Splicing of FN is regulated by several SR proteins, including SRp40 and SRp20, in several different cell types (10)(11)(12). Although the presence of EDB-FN is fairly unique to tumor vasculature, the mechanisms governing its expression within the tumor microenvironment are not clear.…”

mentioning

confidence: 99%

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Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Bordeleau

Califano

Abril

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Alternative splicing of proteins gives rise to different isoforms that play a crucial role in regulating several cellular processes. Notably, splicing profiles are altered in several cancer types, and these profiles are believed to be involved in driving the oncogenic process. Although the importance of alternative splicing alterations occurring during cancer is increasingly appreciated, the underlying regulatory mechanisms remain poorly understood. In this study, we use both biochemical and physical tools coupled with engineered models, patient samples, and a murine model to investigate the role of the mechanical properties of the tumor microenvironment in regulating the production of the extra domain-B (EDB) splice variant of fibronectin (FN), a hallmark of tumor angiogenesis. Specifically, we show that the amount of EDB-FN produced by endothelial cells increases with matrix stiffness both in vitro and within mouse mammary tumors. Matrix stiffness regulates splicing through the activation of serine/arginine rich (SR) proteins, the splicing factors involved in the production of FN isoforms. Activation of the SR proteins by matrix stiffness and the subsequent production of EDB-FN are dependent on intracellular contractility and PI3K-AKT signaling. Notably, matrix stiffness-mediated splicing is not limited to EDB-FN, but also affects splicing in the production of PKC βII and the VEGF 165b splice variant. Together, these results demonstrate that the mechanical properties of the microenvironment regulate alternative splicing and establish a previously unidentified mechanism by which cells can adapt to their microenvironment.alternative splicing | extracellular matrix | matrix stiffness | angiogenesis | cancer progression D ifferential expression of protein isoforms through alternative splicing is a key element to generating protein diversity and can result in widely different cell phenotypes and behaviors (1-3). Interestingly, tumors exhibit several major differences in protein isoform expression patterns compared with healthy tissue (2, 4), and some of these changes are thought to favor oncogenesis (1). Notably, splicing events are regulated at the pre-mRNA level by splicing regulatory factors that include a family of serine/arginine rich (SR) proteins (5). Elevated SR protein levels have been associated with cancer (5); however, the physiological cues that drive splicing are not well defined.Among the various alternatively spliced proteins present in tumors, the splice variant of fibronectin (FN) that includes the extra domain-B (EDB) type III repeat has been of particular interest in the cancer community because inclusion of the EDB fragment has been proposed as a way to identify and target tumor vasculature (6). EDB-FN is produced by endothelial cells (ECs) in tumors and may favor angiogenesis (7-9). Splicing of FN is regulated by several SR proteins, including SRp40 and SRp20, in several different cell types (10-12). Although the presence of EDB-FN is fairly unique to tumor vasculature, the mechanisms governin...

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Section: Ecm Stiffness Regulates Srp40 Phosphorylation In Culture and Inmentioning

confidence: 99%

mentioning

confidence: 99%

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Bordeleau

Califano

Abril

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…A number of morphogenetic factors including hedgehog proteins (Hhgs), Wnt proteins, Notch ligands, members of TGF-β superfamily of growth factors, or FGFs, have been implicated to play important roles in controlling the regulation of these transcription factors in musculoskeletal tissue development. TGF-β1 is among the earliest signals in chondrogenic condensation, and has been shown to stimulate the synthesis of fibronectin, which in turn regulates N-CAM [90]. BMPs play an important role during bone morphogenesis by initiating chondro-progenitor cell determination and differentiation [91].…”

Section: Growth Factors-mentioning

confidence: 99%

“…BMPs play an important role during bone morphogenesis by initiating chondro-progenitor cell determination and differentiation [91]. BMP has also been shown to regulate the later stages of chondrocytes maturation and terminal differentiation to the hypertrophic phenotype [90].…”

Section: Growth Factors-mentioning

confidence: 99%

Controlled differentiation of stem cells

Hwang

Varghese

Elisseeff

2008

Advanced Drug Delivery Reviews

303

217

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The extracellular microenvironment plays a significant role in controlling cellular behavior. Identification of appropriate biomaterials that support cellular attachment, proliferation and, most importantly in the case of human embryonic stem cells, lineage-specific differentiation is critical for tissue engineering and cellular therapy. In addition to growth factors and morphogenetic factors known to induce lineage commitment of stem cells, a number of scaffolding materials, including synthetic and naturally-derived biomaterials, have been utilized in tissue engineering approaches to direct differentiation. This review focuses on recent emerging findings and well-characterized differentiation models of human embryonic stem cells. Additionally, we also discuss about various strategies that have been used in stem cell expansion.

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“…FN expression is increased in areas of cellular condensation that are decreased as cytodifferentiation proceeds (27,28). According to recent studies, isoform switching of Fn mRNA is important for chondrogenic differentiation (29)(30)(31) and that TGF-ß1 treatment during chondrogenic differentiation of ATDC5 cells increases the expression of FN isoforms that lack the extra domainA and B during chondrogenic differentiation of ATDC5 cells (32,33). Regulation of both quantity and quality of FN by TGF-ß1 could be involved in the early stage of chondrogenesis.…”

Section: Discussionmentioning

confidence: 99%

Activation of epidermal growth factor receptor gene is involved in transforming growth factor-β-mediated fibronectin expression in a chondrocyte progenitor cell line

Takeda

Inoue²,

Kutsuna³

et al. 2010

Int J Mol Med

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Abstract. Chondrocytes produce many types of ECM to maintain elasticity and plasticity in articular cartilage of revolute joints. Both transforming growth factor ß (TGF-ß) and bone morphogenetic proteins (BMPs) induce extracellular matrix proteins such as type II·1 collagen and aggrecan during chondrogenic differentiation in vitro. However, differences in the matrix gene expression pattern by the stimulation of TGF-ßs and BMPs remains unclear. In the present study, we created a customized PCR-based ECM array to investigate the pattern of ECM expression genes in the chondrocyte progenitor cell line ATDC5, that was stimulated by TGF-ßs or BMPs. Fibronectin (Fn) expression was drastically induced after TGF-ß stimulation, but not BMP-4. Epidermal growth factor receptor (Egfr) gene was also significantly activated in TGF-ß1-induced chondrogenic differentiation as compared to BMP-4-mediated differentiation. Furthermore, EGFRknockdown assay of the cells showed decreasing Fn expression during TGF-ß1-induced chondrogenic differentiation. These data indicated that Egfr gene activation by TGF-ß is involved in the differences in the expression of cellular matrix genes such as Fn, as compared to the expression pattern induced by BMPs.

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Transforming growth factor-β1 regulates fibronectin isoform expression and splicing factor SRp40 expression during ATDC5 chondrogenic maturation

Cited by 31 publications

References 43 publications

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Controlled differentiation of stem cells

Activation of epidermal growth factor receptor gene is involved in transforming growth factor-β-mediated fibronectin expression in a chondrocyte progenitor cell line

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