Search Results: 1 - 10 of 200 matches for " Toshihisa Komori "
Regulation of bone mass at unloaded condition by osteocyte network
Toshihisa Komori
Arthritis Research & Therapy , 2012, DOI: 10.1186/ar3568
Abstract: We searched for the molecules responsible for disuse osteoporosis using BCL2 transgenic mice. Pyruvate dehydrogenase kinase isozymes (Pdk1, Pdk2, Pdk3, and Pdk4) are negative regulators of pyruvate dehydrogenase complex (PDC), which converts pyruvate to acetyl-CoA in the mitochondria, linking glycolysis to the energetic and anabolic functions of the tricarboxylic acid (TCA) cycle. Pdk4 was upregulated in femurs and tibiae of wild-type mice but not of BCL2 transgenic mice after tail suspension. Bone in Pdk4-/- mice developed normally and was maintained. At unloading, however, bone mass was reduced due to enhanced osteoclastogenesis and Rankl expression in wild-type mice but not in Pdk4-/- mice. Osteoclast differentiation of Pdk4-/- bone marrow-derived monocyte/macrophage lineage cells (BMMs) in the presence of M-CSF and RANKL was suppressed, and osteoclastogenesis was impaired in the coculture of wild-type BMMs and Pdk4-/- osteoblasts, in which Rankl expression and promoter activity were reduced. Further, introduction of Pdk4 into Pdk4-/- BMMs and osteoblasts enhanced osteoclastogenesis and Rankl expression and activated Rankl promoter. These findings indicate that upregulation of Pdk4 expression in osteoblasts and bone marrow cells after unloading is, at least in part, responsible for the enhancement of osteoclastogenesis and bone resorption after unloading [1].
Osteocyte Network; a Negative Regulatory System for Bone Mass Augmented by the Induction of Rankl in Osteoblasts and Sost in Osteocytes at Unloading
Takeshi Moriishi, Ryo Fukuyama, Masako Ito, Toshihiro Miyazaki, Takafumi Maeno, Yosuke Kawai, Hisato Komori, Toshihisa Komori
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0040143
Abstract: Reduced mechanical stress is a major cause of osteoporosis in the elderly, and the osteocyte network, which comprises a communication system through processes and canaliculi throughout bone, is thought to be a mechanosensor and mechanotransduction system; however, the functions of osteocytes are still controversial and remain to be clarified. Unexpectedly, we found that overexpression of BCL2 in osteoblasts eventually caused osteocyte apoptosis. Osteoblast and osteoclast differentiation were unaffected by BCL2 transgene in vitro. However, the cortical bone mass increased due to enhanced osteoblast function and suppressed osteoclastogenesis at 4 months of age, when the frequency of TUNEL-positive lacunae reached 75%. In the unloaded condition, the trabecular bone mass decreased in both wild-type and BCL2 transgenic mice at 6 weeks of age, while it decreased due to impaired osteoblast function and enhanced osteoclastogenesis in wild-type mice but not in BCL2 transgenic mice at 4 months of age. Rankl and Opg were highly expressed in osteocytes, but Rankl expression in osteoblasts but not in osteocytes was increased at unloading in wild-type mice but not in BCL2 transgenic mice at 4 months of age. Sost was locally induced at unloading in wild-type mice but not in BCL2 transgenic mice, and the dissemination of Sost was severely interrupted in BCL2 transgenic mice, showing the severely impaired osteocyte network. These findings indicate that the osteocyte network is required for the upregulation of Rankl in osteoblasts and Sost in osteocytes in the unloaded condition. These findings suggest that the osteocyte network negatively regulate bone mass by inhibiting osteoblast function and activating osteoclastogenesis, and these functions are augmented in the unloaded condition at least partly through the upregulation of Rankl expression in osteoblasts and that of Sost in osteocytes, although it cannot be excluded that low BCL2 transgene expression in osteoblasts contributed to the enhanced osteoblast function.
Bcl2 Deficiency Activates FoxO through Akt Inactivation and Accelerates Osteoblast Differentiation
Takeshi Moriishi, Yosuke Kawai, Hisato Komori, Satoshi Rokutanda, Yutaka Eguchi, Yoshihide Tsujimoto, Izumi Asahina, Toshihisa Komori
Abstract: Osteoblast apoptosis plays an important role in bone development and maintenance, and is in part responsible for osteoporosis in sex steroid deficiency, glucocorticoid excess, and aging. Although Bcl2 subfamily proteins, including Bcl2 and Bcl-XL, inhibit apoptosis, the physiological significance of Bcl2 in osteoblast differentiation has not been fully elucidated. To investigate this, we examined Bcl2-deficient (Bcl2?/?) mice. In Bcl2?/? mice, bromodeoxyuridine (BrdU)-positive osteoblasts were reduced in number, while terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive osteoblasts were increased. Unexpectedly, osteoblast differentiation was accelerated in Bcl2?/? mice as shown by the early appearance of osteocalcin-positive osteoblasts. Osteoblast differentiation was also accelerated in vitro when primary osteoblasts were seeded at a high concentration to minimize the reduction of the cell density by apoptosis during culture. FoxO transcription factors, whose activities are negatively regulated through the phosphorylation by Akt, play important roles in multiple cell events, including proliferation, death, differentiation, longevity, and stress response. Expressions of FasL, Gadd45a, and Bim, which are regulated by FoxOs, were upregulated; the expression and activity of FoxOs were enhanced; and the phosphorylation of Akt and that of FoxO1 and FoxO3a by Akt were reduced in Bcl2?/? calvariae. Further, the levels of p53 mRNA and protein were increased, and the expression of p53-target genes, Pten and Igfbp3 whose proteins inhibit Akt activation, was upregulated in Bcl2?/? calvariae. However, Pten but not Igfbp3 was upregulated in Bcl2?/? primary osteoblasts, and p53 induced Pten but not Igfbp3 in vitro. Silencing of either FoxO1 or FoxO3a inhibited and constitutively-active FoxO3a enhanced osteoblast differentiation. These findings suggest that Bcl2 deficiency induces and activates FoxOs through Akt inactivation, at least in part, by upregulating Pten expression through p53 in osteoblasts, and that the enhanced expression and activities of FoxOs may be one of the causes of accelerated osteoblast differentiation in Bcl2?/? mice.
Thrombospondin-1 Is a Putative Target Gene of Runx2 and Runx3
Xiuming Shi,Vishwa Deepak,Linghui Wang,Xueqing Ba,Toshihisa Komori,Xianlu Zeng,Wenguang Liu
International Journal of Molecular Sciences , 2013, DOI: 10.3390/ijms140714321
Abstract: Thrombospondin-1 (TSP-1), a matricellular protein widely acclaimed to be involved in the inhibition of angiogenesis and tumorigenesis, is synthesized and secreted by many cell types, including osteoblast and cancer cells. TSP-1 is highly upregulated during early stage of osteogenesis, whereas it inhibits terminal osteoblast differentiation. Expression of TSP-1 is downregulated in cancer cells, and its ectopic expression has been shown to restrain tumor growth. Transcriptional regulation of TSP-1 in osteogenesis and cancer is poorly understood; this prompted us to study its regulation by the two key regulators of the aforementioned processes: Runx2 and Runx3. Through a PCR-based cDNA subtraction technique, we identified and cloned a cDNA fragment for mouse TSP-1, whose expression was dramatically upregulated in response to Runx2 expression in mesenchymal stem cells. Moreover, TSP-1 expression was considerably reduced in the lung of Runx2 knockout mouse. On the other hand, TSP-1 gene expression drastically increased at both the transcriptional and translational levels in response to Runx3 expression in B16-F10 melanoma cells. In line with this, Runx2 and Runx3 bound to the TSP-1 promoter and stimulated its activity. Hence, these results provide first line of evidence that TSP-1 is a transcriptional target gene of Runx2 and Runx3.
Overexpression of Bcl2 in Osteoblasts Inhibits Osteoblast Differentiation and Induces Osteocyte Apoptosis
Takeshi Moriishi, Zenjiro Maruyama, Ryo Fukuyama, Masako Ito, Toshihiro Miyazaki, Hideki Kitaura, Hidetake Ohnishi, Tatsuya Furuichi, Yosuke Kawai, Ritsuko Masuyama, Hisato Komori, Kenji Takada, Hiroshi Kawaguchi, Toshihisa Komori
Abstract: Bcl2 subfamily proteins, including Bcl2 and Bcl-XL, inhibit apoptosis. As osteoblast apoptosis is in part responsible for osteoporosis in sex steroid deficiency, glucocorticoid excess, and aging, bone loss might be inhibited by the upregulation of Bcl2; however, the effects of Bcl2 overexpression on osteoblast differentiation and bone development and maintenance have not been fully investigated. To investigate these issues, we established two lines of osteoblast-specific BCL2 transgenic mice. In BCL2 transgenic mice, bone volume was increased at 6 weeks of age but not at 10 weeks of age compared with wild-type mice. The numbers of osteoblasts and osteocytes increased, but osteoid thickness and the bone formation rate were reduced in BCL2 transgenic mice with high expression at 10 weeks of age. The number of BrdU-positive cells was increased but that of TUNEL-positive cells was unaltered at 2 and 6 weeks of age. Osteoblast differentiation was inhibited, as shown by reduced Col1a1 and osteocalcin expression. Osteoblast differentiation of calvarial cells from BCL2 transgenic mice also fell in vitro. Overexpression of BCL2 in primary osteoblasts had no effect on osteoclastogenesis in co-culture with bone marrow cells. Unexpectedly, overexpression of BCL2 in osteoblasts eventually caused osteocyte apoptosis. Osteocytes, which had a reduced number of processes, gradually died with apoptotic structural alterations and the expression of apoptosis-related molecules, and dead osteocytes accumulated in cortical bone. These findings indicate that overexpression of BCL2 in osteoblasts inhibits osteoblast differentiation, reduces osteocyte processes, and causes osteocyte apoptosis.
MAML1 Enhances the Transcriptional Activity of Runx2 and Plays a Role in Bone Development
Takashi Watanabe,Toshinao Oyama,Maki Asada,Daisuke Harada,Yoshiaki Ito,Masayo Inagawa,Yutaka Suzuki,Sumio Sugano,Ken-ichi Katsube,Gerard Karsenty,Toshihisa Komori,Motoo Kitagawa,Hiroshi Asahara
PLOS Genetics , 2013, DOI: 10.1371/journal.pgen.1003132
Abstract: Mastermind-like 1 (MAML1) is a transcriptional co-activator in the Notch signaling pathway. Recently, however, several reports revealed novel and unique roles for MAML1 that are independent of the Notch signaling pathway. We found that MAML1 enhances the transcriptional activity of runt-related transcription factor 2 (Runx2), a transcription factor essential for osteoblastic differentiation and chondrocyte proliferation and maturation. MAML1 significantly enhanced the Runx2-mediated transcription of the p6OSE2-Luc reporter, in which luciferase expression was controlled by six copies of the osteoblast specific element 2 (OSE2) from the Runx2-regulated osteocalcin gene promoter. Interestingly, a deletion mutant of MAML1 lacking the N-terminal Notch-binding domain also enhanced Runx2-mediated transcription. Moreover, inhibition of Notch signaling did not affect the action of MAML1 on Runx2, suggesting that the activation of Runx2 by MAML1 may be caused in a Notch-independent manner. Overexpression of MAML1 transiently enhanced the Runx2-mediated expression of alkaline phosphatase, an early marker of osteoblast differentiation, in the murine pluripotent mesenchymal cell line C3H10T1/2. MAML1?/? embryos at embryonic day 16.5 (E16.5) had shorter bone lengths than wild-type embryos. The area of primary spongiosa of the femoral diaphysis was narrowed. At E14.5, extended zone of collagen type II alpha 1 (Col2a1) and Sox9 expression, markers of chondrocyte differentiation, and decreased zone of collagen type X alpha 1 (Col10a1) expression, a marker of hypertrophic chondrocyte, were observed. These observations suggest that chondrocyte maturation was impaired in MAML1?/? mice. MAML1 enhances the transcriptional activity of Runx2 and plays a role in bone development.
SP7 Inhibits Osteoblast Differentiation at a Late Stage in Mice
Carolina A. Yoshida,Hisato Komori,Zenjiro Maruyama,Toshihiro Miyazaki,Keishi Kawasaki,Tatsuya Furuichi,Ryo Fukuyama,Masako Mori,Kei Yamana,Kouhei Nakamura,Wenguang Liu,Satoru Toyosawa,Takeshi Moriishi,Hiroshi Kawaguchi,Kenji Takada,Toshihisa Komori
Abstract: RUNX2 and SP7 are essential transcription factors for osteoblast differentiation at an early stage. Although RUNX2 inhibits osteoblast differentiation at a late stage, the function of SP7 at the late stage of osteoblast differentiation is not fully elucidated. Thus, we pursued the function of SP7 in osteoblast differentiation. RUNX2 induced Sp7 expression in Runx2?/? calvarial cells. Adenoviral transfer of sh-Sp7 into primary osteoblasts reduced the expression of Alpl, Col1a1, and Bglap2 and mineralization, whereas that of Sp7 reduced Bglap2 expression and mineralization at a late stage of osteoblast differentiation. Sp7 transgenic mice under the control of 2.3 kb Col1a1 promoter showed osteopenia and woven-bone like structure in the cortical bone, which was thin and less mineralized, in a dose-dependent manner. Further, the number of processes in the osteoblasts and osteocytes was reduced. Although the osteoblast density was increased, the bone formation was reduced. The frequency of BrdU incorporation was increased in the osteoblastic cells, while the expression of Col1a1, Spp1, Ibsp, and Bglap2 was reduced. Further, the osteopenia in Sp7 or Runx2 transgenic mice was worsened in Sp7/Runx2 double transgenic mice and the expression of Col1a1 and Bglap2 was reduced. The expression of Sp7 and Runx2 was not increased in Runx2 and Sp7 transgenic mice, respectively. The expression of endogenous Sp7 was increased in Sp7 transgenic mice and Sp7-transduced cells; the introduction of Sp7 activated and sh-Sp7 inhibited Sp7 promoter; and ChIP assay showed the binding of endogenous SP7 in the proximal region of Sp7 promoter. These findings suggest that SP7 and RUNX2 inhibit osteoblast differentiation at a late stage in a manner independent of RUNX2 and SP7, respectively, and SP7 positively regulates its own promoter.
A System of Third-Order Differential Operators Conformally Invariant under $\mathfrak{so}(8,\mathbb{C})$
Toshihisa Kubo
Abstract: In earlier work, Barchini, Kable, and Zierau constructed a number of conformally invariant systems of differential operators associated to Heisenberg parabolic subalgebras in simple Lie algebras. The construction was systematic, but the existence of such a system was left open in several anomalous cases. Here, a conformally invariant system is shown to exist in the most interesting of these remaining cases. The construction may also be interpreted as giving an explicit homomorphism between generalized Verma modules for the Lie algebra of type $D_4$.
Special Values for Conformally Invariant Systems Associated to Maximal Parabolics of Quasi-Heisenberg Type
Abstract: In this paper we construct conformally invariant systems of first order and second order differential operators associated to a homogeneous line bundle $\Cal{L}_{s} \to G_0/Q_0$ with $Q_0$ a maximal parabolic subgroup of quasi-Heisenberg type. This generalizes the results by Barchini, Kable, and Zierau.To do so we use techniques different from ones used by them.
Stability of multidimensional skip-free Markov modulated reflecting random walks: Revisit to Malyshev and Menshikov's results and application to queueing networks
Toshihisa Ozawa
Abstract: Let $\{\boldsymbol{X}_n\}$ be a discrete-time $d$-dimensional process on $\mathbb{Z}_+^d$ with a supplemental (background) process $\{J_n\}$ on a finite set and assume the joint process $\{\boldsymbol{Y}_n\}=\{(\boldsymbol{X}_n,J_n)\}$ to be Markovian. Then, the process $\{\boldsymbol{X}_n\}$ can be regarded as a kind of reflecting random walk (RRW for short) in which the transition probabilities of the RRW are modulated according to the state of the background process $\{J_n\}$; we assume this modulation is space-homogeneous inside $\mathbb{Z}_+^d$ and on each boundary face of $\mathbb{Z}_+^d$. Further we assume the process $\{\boldsymbol{X}_n\}$ is skip free in all coordinates and call the joint process $\{\boldsymbol{Y}_n\}$ a $d$-dimensional skip-free Markov modulated reflecting random walk (MMRRW for short). The MMRRW is an extension of an ordinary RRW and stability of ordinary RRWs have been studied by Malyshev and Menshikov. Following their results, we obtain stability and instability conditions for MMRRWs and apply our results to stability analysis of a two-station network.