Efficient Expression of a Heterodimer of Bone Morphogenetic Protein Subunits Using a Baculovirus Expression System

Hazama, Masatoshi; Aono, Aki; Ueno, Naoto; Fujisawa, Yukio

doi:10.1006/bbrc.1995.1578

Cited by 112 publications

(76 citation statements)

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“…In vitro studies have shown that co-expression of BMP2 and BMP7 can lead to the formation of biologically active heterodimeric BMP2/7 molecules (Hazama et al, 1995;Israel et al, 1996;Zhao et al, 2005). Although the physiological role of heterodimeric BMP molecules still awaits validation in vivo, it has been shown that forced expression or treatment with BMP2/7 heterodimers potently stimulated bone formation in vitro as well as in vivo (Hazama et al, 1995;Israel et al, 1996;Zhu et al, 2004;Zhao et al, 2005), thus providing a rationale for the development of BMP heterodimeric molecules for clinical use in bone fracture healing.…”

Section: Bmp2/7 Inhibits the Breast Cancer Stem Cell Subpopulation Jtmentioning

confidence: 99%

“…Although the physiological role of heterodimeric BMP molecules still awaits validation in vivo, it has been shown that forced expression or treatment with BMP2/7 heterodimers potently stimulated bone formation in vitro as well as in vivo (Hazama et al, 1995;Israel et al, 1996;Zhu et al, 2004;Zhao et al, 2005), thus providing a rationale for the development of BMP heterodimeric molecules for clinical use in bone fracture healing. In line with these observations, a recent publication on a BMP2/6 heterodimer also demonstrated the superiority of the heterodimer over its homodimeric BMP2 or BMP6 counterparts in the differentiation induction of human embryonic stem cells in vitro (Valera et al, 2010).…”

Section: Bmp2/7 Inhibits the Breast Cancer Stem Cell Subpopulation Jtmentioning

confidence: 99%

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The BMP2/7 heterodimer inhibits the human breast cancer stem cell subpopulation and bone metastases formation

et al. 2011

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Accumulating evidence suggests that a subpopulation of breast cancer cells, referred to as cancer stem cells (CSCs), have the ability to propagate a tumor and potentially seed new metastases. Furthermore, stimulation of an epithelialto-mesenchymal transition by factors like transforming growth factor-b (TGFb) is accompanied with the generation of breast CSCs. Previous observations indicated that bone morphogenetic protein-7 (BMP7) antagonizes the protumorigenic and prometastatic actions of TGFb, but whether BMP7 action is mechanistically linked to breast CSCs has remained elusive. Here, we have studied the effects of BMP7, BMP2 and a BMP2/7 heterodimer on the formation of human breast CSCs (ALDH hi /CD44 hi / CD24 À/low ) and bone metastases formation in a preclinical model of intra-cardiac injection of MDA-MB-231 cells in athymic nude (Balb/c nu/nu) mice. The BMP2/7 heterodimer was the most efficient stimulator of BMP signaling and very effectively reduced TGFb-driven Smad signaling and cancer cell invasiveness. The tested BMPs-particularly the heterodimeric BMP2/7-strongly reduced the size of the ALDH hi /CD44 hi /CD24 À/low CSC subpopulation. In keeping with these in vitro observations, pretreatment of cancer cells with BMPs for 72 h prior to systemic inoculation of the cancer cells inhibited the formation of bone metastases. Collectively, our data support the notion that breast CSCs are involved in bone metastasis formation and describe heterodimeric BMP2/7 as a powerful TGFb antagonist with anti-metastatic potency.

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Section: Bmp2/7 Inhibits the Breast Cancer Stem Cell Subpopulation Jtmentioning

confidence: 99%

Section: Bmp2/7 Inhibits the Breast Cancer Stem Cell Subpopulation Jtmentioning

confidence: 99%

The BMP2/7 heterodimer inhibits the human breast cancer stem cell subpopulation and bone metastases formation

et al. 2011

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“…The BMP-7 vector induced only weak alkaline phosphatase activity. 35,36 Recombinant BMP adenoviruses have been the most commonly used vectors in BMP gene therapy. Direct injection of recombinant BMP-2 adenovirus can induce bone formation in immunodefective and immunocompetent mice and athymic nude rats, but could not stimulate ectopic bone formation in Sprague-Dawley rats.…”

Section: Introductionmentioning

confidence: 99%

Osteogenic potential of five different recombinant human bone morphogenetic protein adenoviral vectors in the rat

et al. 2003

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one morphogenetic protein (BMP) adenoviral vectors for the induction of osteogenesis are being developed for the treatment of bone pathology. However, it is still unknown which BMP adenoviral vector has the highest potential to stimulate bone formation in vivo. In this study, the osteogenic activities of recombinant human BMP-2, BMP-4, BMP-6, BMP-7, and BMP-9 adenoviruses were compared in vitro, in athymic nude rats, and in Sprague-Dawley rats. In vitro osteogenic activity was assessed by measuring the alkaline phosphatase activity in C2C12 cells transduced by the various BMP vectors. The alkaline phosphatase activity induced by 2 Â 10 5 PFU/well of BMP viral vector was 4890 Â 10 À12 U/well for ADCMVBMP-9, 302 Â 10 À12 U/well for ADCMVBMP-4, 220 Â 10 À12 U/well for ADCMVBMP-6, 45 Â 10 À12 U/well for ADCMVBMP-2, and 0.43 Â 10 À12 U/ well for ADCMVBMP-7. The average volume of new bone induced by 10 7 PFU of BMP vector in athymic nude rats was 0.3770.03 cm 3 for ADCMVBMP-2, 0.8970.07 cm 3 for ADCMVBMP-4, 1.0270.07 cm 3 for ADCMVBMP-6, 0.2470.05 cm 3 for ADCMVBMP-7, and 0.6370.07 cm 3 for ADCMVBMP-9. In immunocompetent Sprague-Dawley rats, no bone formation was demonstrated in the ADCMVBMP-2, ADCMVBMP-4, and ADCMVBMP-7 groups. ADCMVBMP-6 at a viral dose of 10 8 PFU induced 0.1070.03 cm 3 of new bone, whereas ADCMVBMP-9 at a lower viral dose of 10 7 PFU induced more bone, with an average volume of 0.2970.01 cm 3 .

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“…Heterodimers composed of distinct BMP family members show a higher specific activity than do homodimers of either subunit. Homodimers of BMP2, -4, or -7, for example, can all induce bone formation, but heterodimers of BMP2 plus BMP7, or BMP4 plus BMP7 are significantly more potent (5-to 20-fold) than any of the homodimers in osteogenic differentiation assays (4)(5)(6). BMP2/7 and BMP4/7 heterodimers also show enhanced activity in Xenopus and zebrafish embryo assays (7,8).…”

mentioning

confidence: 99%

The prodomain of BMP4 is necessary and sufficient to generate stable BMP4/7 heterodimers with enhanced bioactivity in vivo

Neugebauer¹,

Kwon

Kim³

et al. 2015

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

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Bone morphogenetic proteins 4 and 7 (BMP4 and BMP7) are morphogens that signal as either homodimers or heterodimers to regulate embryonic development and adult homeostasis. BMP4/7 heterodimers exhibit markedly higher signaling activity than either homodimer, but the mechanism underlying the enhanced activity is unknown. BMPs are synthesized as inactive precursors that dimerize and are then cleaved to generate both the bioactive ligand and prodomain fragments, which lack signaling activity. Our study reveals a previously unknown requirement for the BMP4 prodomain in promoting heterodimer activity. We show that BMP4 and BMP7 precursor proteins preferentially or exclusively form heterodimers when coexpressed in vivo. In addition, we show that the BMP4 prodomain is both necessary and sufficient for generation of stable heterodimeric ligands with enhanced activity and can enable homodimers to signal in a context in which they normally lack activity. Our results suggest that intrinsic properties of the BMP4 prodomain contribute to the relative bioactivities of homodimers versus heterodimers in vivo. These findings have clinical implications for the use of BMPs as regenerative agents for the treatment of bone injury and disease.B one morphogenetic proteins (BMPs) are members of the TGFβ superfamily that were originally isolated as boneinducing morphogens and were subsequently found to play central roles during embryogenesis and in adult homeostasis (1). BMPs are clinically important therapeutic agents that are used to reverse bone loss caused by trauma, disease, and tumor resection (2). Their use as regenerative agents is limited, however, by their short half-life and low specific activity when implanted in vivo. Understanding how BMP activity is regulated is important for the development of more effective therapeutic agents for the treatment of bone injury and disease.BMPs bind to and activate a receptor complex consisting of type I and type II transmembrane serine/threonine kinases. Following ligand binding, activated receptors propagate their signal by phosphorylating one of the SMADs that is specific for the BMP pathway (SMAD1, -5, or -8). The phosphorylated Smads then form heterooligomers with the common Smad, Smad4, and this complex translocates into the nucleus where it binds to BMP response elements and activates transcription of target genes (1).BMPs are classified into subfamilies based on sequence homology. They signal as either homodimers, or as heterodimers from different subfamilies. For example, class I BMPs, which consist of BMP2 and BMP4, can heterodimerize with class II BMPs, consisting of BMP5-8 (3). Heterodimers composed of distinct BMP family members show a higher specific activity than do homodimers of either subunit. Homodimers of BMP2, -4, or -7, for example, can all induce bone formation, but heterodimers of BMP2 plus BMP7, or BMP4 plus BMP7 are significantly more potent (5-to 20-fold) than any of the homodimers in osteogenic differentiation assays (4-6). BMP2/7 and BMP4/7 heterodimers also sho...

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Efficient Expression of a Heterodimer of Bone Morphogenetic Protein Subunits Using a Baculovirus Expression System

Cited by 112 publications

References 0 publications

The BMP2/7 heterodimer inhibits the human breast cancer stem cell subpopulation and bone metastases formation

The BMP2/7 heterodimer inhibits the human breast cancer stem cell subpopulation and bone metastases formation

Osteogenic potential of five different recombinant human bone morphogenetic protein adenoviral vectors in the rat

The prodomain of BMP4 is necessary and sufficient to generate stable BMP4/7 heterodimers with enhanced bioactivity in vivo

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