Inactivation of Menin, the Product of the Multiple Endocrine Neoplasia Type 1 Gene, Inhibits the Commitment of Multipotential Mesenchymal Stem Cells into the Osteoblast Lineage

Sowa, Hideaki; Kaji, Hiroshi; Canaff, Lucie; Hendy, Geoffrey N.; Tsukamoto, Tatsuo; Yamaguchi, Toru; Miyazono, Kohei; Sugimoto, Toshitsugu; Chihara, Kazuo

doi:10.1074/jbc.m302044200

Cited by 89 publications

(107 citation statements)

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“…As we reported previously (14), ALP activity increases with time, and the expression of type 1 collagen and osteopontin peaks at 7 and 14 days of culture, respectively, whereas mRNA expression of OCN, a terminal differentiation marker, is highest at day 21 of culture. The levels of Tmem119 mRNA increased for up to 2 weeks, and the level was maintained up to week 3 in MC3T3-E1 cells (Fig.…”

Section: Tmem119 Expression During Osteoblast Differentiation-supporting

confidence: 83%

“…Transient or Stable Transfection-Each vector was transfected into MC3T3-E1, ST-2, COS-7, or C2C12 cells with Lipofectamine (Invitrogen) as described previously (13)(14)(15). Six h later, the cells were fed fresh ␣-MEM containing 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

“…Reverse transcription of 5 g of cultured cell total RNA was carried out for 50 min at 42°C and then for 15 min at 70°C using the SuperScript First Strand Synthesis System for RT-PCR (Invitrogen), which contained RT buffer, oligo(deoxythymidine) [12][13][14][15][16][17][18] , 5ϫ first-strand solution, 10 mM deoxy-NTP, 0.1 M dithiothreitol, SuperScript II (RT enzyme), and ribonuclease H (ribonuclease inhibitor). PCR, using primers to unique sequences in each cDNA, was carried out in a volume of 10 l of PCR reaction mixture (supplied by TaKaRa, Otsu, Japan) supplemented with 2.5 units of TaKaRa Taq, deoxy-NTP (1.5 mM each), and PCR buffer (10ϫ; 100 mM Tris-HCl (pH 8.3), 500 mM KCl, and 15 mM MgCl 2 ); 25 ng of each primer and 1 l of template (from a 50-l RT reaction) were used.…”

Section: Methodsmentioning

confidence: 99%

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Parathyroid Hormone-responsive Smad3-related Factor, Tmem119, Promotes Osteoblast Differentiation and Interacts with the Bone Morphogenetic Protein-Runx2 Pathway

Hisa

Inoue

Hendy

et al. 2011

Journal of Biological Chemistry

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The mechanisms whereby the parathyroid hormone (PTH) exerts its anabolic action on bone are incompletely understood. We previously showed that inhibition of ERK1/2 enhanced Smad3-induced bone anabolic action in osteoblasts. These findings suggested the hypothesis that changes in gene expression associated with the altered Smad3-induced signaling brought about by an ERK1/2 inhibitor would identify novel bone anabolic factors in osteoblasts. We therefore performed a comparative DNA microarray analysis between empty vector-transfected mouse osteoblastic MC3T3-E1 cells and PD98059-treated stable Smad3-overexpressing MC3T3-E1 cells. Among the novel factors, Tmem119 was selected on the basis of its rapid induction by PTH independent of later increases in endogenous TGF-␤. The levels of Tmem119 increased with time in cultures of MC3T3-E1 cells and mouse mesenchymal ST-2 cells committed to the osteoblast lineage by BMP-2. PTH stimulated Tmem119 levels within 1 h as determined by Western blot analysis and immunocytochemistry in MC3T3-E1 cells. MC3T3-E1 cells stably overexpressing Tmem119 exhibited elevated levels of Runx2, osteocalcin, alkaline phosphatase, and ␤-catenin, whereas Tmem119 augmented BMP-2-induced Runx2 levels in mesenchymal cells. Tmem119 interacted with Runx2, Smad1, and Smad5 in C2C12 cells. In conclusion, we identified a Smad3-related factor, Tmem119, that is induced by PTH and promotes differentiation in mouse osteoblastic cells. Tmem119 is an important molecule in the pathway downstream of PTH and Smad3 signaling in osteoblasts.Bone remodeling is controlled by bone resorption and subsequent osteoblastic bone formation. Insights into the mechanisms of osteoclastic bone resorption have led to the development of reagents that potently suppress bone resorption for the treatment of osteoporosis. On the other hand, the impairment of osteoblastic bone formation is considered to be the main driving force in age-related, glucocorticoid-induced, and diabetes-related osteoporosis. However, many aspects of osteoblastic bone formation remain to be understood. Intermittent parathyroid hormone (PTH) 3 is one of the most potent anabolic agents; it stimulates de novo osteoblastic bone formation and is superior to bisphosphonates in the treatment of osteoporosis (1, 2). The anabolic action of PTH is exerted partly through local growth factors and transcriptional regulators as well by as an anti-apoptotic action in osteoblasts (3, 4). However, the precise mechanisms by which PTH exerts its anabolic action on bone are incompletely understood.We showed previously that Smad3, a crucial TGF-␤-signaling molecule, promotes alkaline phosphatase (ALP) activity in mouse osteoblastic MC3T3-E1 cells (5, 6), and Borton et al. (7) have found that mice with targeted disruption of Smad3 exhibit osteopenia caused by decreased bone formation, suggesting that the Smad3 molecule is a promoter of bone formation. Moreover, we demonstrated that TGF-␤-responsive ERK1/2 and c-Jun N-terminal kinase (JNK) cascades negatively regulate Smad3-ind...

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Section: Tmem119 Expression During Osteoblast Differentiation-supporting

confidence: 83%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Parathyroid Hormone-responsive Smad3-related Factor, Tmem119, Promotes Osteoblast Differentiation and Interacts with the Bone Morphogenetic Protein-Runx2 Pathway

Hisa

Inoue

Hendy

et al. 2011

Journal of Biological Chemistry

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“…Although the precise biochemical function of menin in this complex needs to be elucidated, the importance of menin in this complex is apparent from the observation that the loss of menin results in reduced H3K4 methylation and associated reduction in the expression of known mixed lineage leukemia targets Hoxc6 and Hoxc8 (6). Interaction of menin with Smad3 is reported to result in activation of transforming growth factor-h (TGF-h)-induced transcription (8), and the interaction of menin with Smad1, Smad3, Smad5, and Runx2 may participate in the TGF-hmediated and bone morphogenetic protein-2 -mediated pathways of commitment of multipotential mesenchymal stem cells to osteoblast lineage (9)(10)(11). Previous studies of gene expression in tumors, cell lines, and embryos have identified nonoverlapping sets of gene(s), whose expression is affected by menin (5,12,13).…”

Section: Introductionmentioning

confidence: 99%

Mouse Embryo Fibroblasts Lacking the Tumor Suppressor Menin Show Altered Expression of Extracellular Matrix Protein Genes

Ji¹,

Prasad²,

Novotny³

et al. 2007

Molecular Cancer Research

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Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant familial cancer syndrome characterized primarily by endocrine tumors of the parathyroids, anterior pituitary, and enteropancreatic endocrine tissues. Affected individuals carry a germ-line loss-of-function mutation of the MEN1 gene, and tumors arise after loss of the second allele. Homozygous loss of Men1 in the germ line of mice results in early embryonic lethality, with defective development of neural tube, heart, liver, and craniofacial structures. We generated immortalized wild-type (WT) and menin-null mouse embryo fibroblast (MEF) cell lines and evaluated their characteristics, including global expression patterns. The WT and menin-null cell lines were aneuploid, and the nulls did not display tumorigenic characteristics in soft agar assay. Expression arrays in menin-null MEFs revealed altered expression of several extracellular matrix proteins that are critical in organogenesis. Specifically, transcripts for fibulin 2 (Fbln2), periostin (Postn), and versican [chondroitin sulfate proteoglycan (Cspg2)], genes critical for the developing heart and known to be induced by transforming growth factor-B (TGF-B), were decreased in their expression in menin-null MEFs. Fbln2 expression was the most affected, and the reduction in menin-null MEFs for Fbln2, Postn, and Cspg2 was 16.18-, 5.37-, and 2.15-fold, respectively. Menin-null MEFs also showed poor response to TGF-B -induced Smad3-mediated transcription in a reporter assay, supporting a role for menin in this pathway. Postn and Cspg2 expression in WT, unlike in null MEFs, increased on TGF-B treatment. The expression changes associated with the loss of the tumor suppressor menin provide insights into the defective organogenesis observed during early embryonic development in Men1-null mouse embryos.

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“…Numerous studies have aimed to elucidate menin's function, primarily through investigating its molecular interactions (for a review see Poisson et al, 2003). Menin binds to a variety of transcription factors (Heppner et al, 2001;Kaji et al, 2001;Chandrasekharappa and Teh, 2003;Sowa et al, 2003). It has been found to repress JunD-activated transcription (Gobl et al, 1999;Kim et al, 2003), revealing that, when deprived of menin, JunD switches from growth suppressor to growth promoter (Agarwal et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Lack of augmentation of tumor spectrum or severity in dual heterozygous Men1 and Rb1 knockout mice

et al. 2006

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To identify possible genetic interactions between the mechanisms of tumor suppression of menin and pRb, we intercrossed mice with targeted deletions of Men1 and Rb1, and compared tumor development in cohorts of animals carrying single or dual mutations of these tumorsuppressor genes. In mice lacking one copy of Men1, pancreatic islet and anterior pituitary adenomas are common. In animals lacking one copy of Rb1, intermediate pituitary and thyroid tumors occur at high frequency, with less frequent development of pancreatic islet hyperplasia and parathyroid lesions. In mice heterozygous for both Men1 and Rb1, pancreatic hyperplasia and tumors of the intermediate pituitary and thyroid occurred at high frequency. Serum measurements of calcium and glucose did not vary significantly between genotypic groups. Loss of heterozygosity at the Rb1 locus was common in pituitary and thyroid tumors, whereas loss of menin was observed in pancreatic and parathyroid lesions. The tumor spectrum in the double heterozygotes was a combination of pathologies seen in each of the individual heterozygotes, without decrease in age of onset, indicating independent, non-additive effects of the two mutations. Together with the lack of increased tumor spectrum, this suggests that menin and pRb function in a common pathway of tumor suppression.

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Inactivation of Menin, the Product of the Multiple Endocrine Neoplasia Type 1 Gene, Inhibits the Commitment of Multipotential Mesenchymal Stem Cells into the Osteoblast Lineage

Abstract: The physiological roles of menin, the product of the multiple endocrine neoplasia type 1 gene, are not known. Homozygous menin knockout mice exhibit cranial and facial hypoplasia. We, therefore, investigated the role of menin in the regulation of osteoblastic differ-

Cited by 89 publications

References 56 publications

Parathyroid Hormone-responsive Smad3-related Factor, Tmem119, Promotes Osteoblast Differentiation and Interacts with the Bone Morphogenetic Protein-Runx2 Pathway

Parathyroid Hormone-responsive Smad3-related Factor, Tmem119, Promotes Osteoblast Differentiation and Interacts with the Bone Morphogenetic Protein-Runx2 Pathway

Mouse Embryo Fibroblasts Lacking the Tumor Suppressor Menin Show Altered Expression of Extracellular Matrix Protein Genes

Lack of augmentation of tumor spectrum or severity in dual heterozygous Men1 and Rb1 knockout mice

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