THE PHYSIOLOGY OF IGFBP DEGRADING PROTEINASES IN BONE

IGFBP 降解骨中蛋白酶的生理学

基本信息

批准号：
7650176
负责人：
John L Fowlkes
金额：
$ 24.36万
依托单位：
ARKANSAS CHILDREN'S HOSPITAL RES INST
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-06-15 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7650176
关键词：
Affect Affinity Anabolic Agents Binding Binding Proteins Biological Biological Availability Cell Line Cell surface Complex Data Differentiation and Growth Distraction Osteogenesis Eosinophil Major Basic Protein Equilibrium FDA approved Family Gene Expression Hormones In Vitro Insulin-Like Growth Factor Binding Protein 4 Insulin-Like Growth Factor Binding Protein 5 Insulin-Like Growth Factor I Insulin-Like Growth Factor II Insulin-Like Growth Factor Receptor Insulin-Like Growth-Factor-Binding Proteins Insulin-Like-Growth Factor I Receptor Intervention Laboratories Lead Matrix Metalloproteinases Mediating Mediator of activation protein Metalloproteases Modeling Osteoblasts Osteogenesis Parathyroid Hormones Peptide Hydrolases Physiology Play Pregnancy-Associated Plasma Protein-A Process Production Proteolysis Proteolytic Processing Reagent Recombinants Resistance Role Signal Transduction Small Interfering RNA Somatomedins Stream System Technology Therapeutic Intervention Transgenic Organisms Up-Regulation bone bone cell bone growth factor bone loss collagenase 3 human PTH protein in vivo inhibitor/antagonist insight member mutant osteoblast differentiation overexpression proteinase In

项目摘要

DESCRIPTION (provided by applicant): Insulin-like growth factors (IGFs) are important mediators of growth and differentiation of osteoblasts; however, six high-affinity IGF-binding proteins (IGFBPs 1-6) antagonize their binding to cell surface type-1 IGF receptors. Proteases produced by bone cells can degrade IGFBPs into low-affinity fragments, allowing for IGF-IGF receptor interactions to occur. We have identified pregnancy-associated plasma protein-A (PAPP-A) as an IGFBP-4 proteinase and MMP-13 (collagenase-3) as an IGFBP-5 proteinase, and both are produced by the osteoblast cell line, MC3T3-E1. Thus, proteolytic processing of these IGFBPs may modulate IGF action during osteoblastogenesis. We have created recombinant mutant forms of IGFBP-4 and IGFBP-5 that are resistant to proteolysis, and have used these constructs to establish stably transfected MC3T3-E1 cell lines overexpressing protease resistant IGFBP mutants. Herein, we propose to use these reagents in order to explore how IGFBP proteolysis influences osteoblast growth and differentiation. We will compare how protease-resistant forms of IGFBP-4 or -5 impact osteoblastogenesis compared to cleavable forms, and if inhibiting or down-regulating IGFBP-degrading proteinases (i.e., PAPP-A and MMP-13) influences this process. In vivo, we will examine how protease-resistant or sensitive forms of IGFBP-4 or -5 alter new bone formation, and if PAPP-A can overcome osteoinhibitory effects of transgenic overproduction of IGFBP-4 on new bone formation. To explore how MMP-13 may mediate anabolic affects of PTH on bone cells, we will determine how blunting of PTH-mediated up-regulation of MMP-13 may alter IGF action in osteoblasts in vitro. We will also explore how MMP-13 resistant forms of IGFBP-5 impact PTH actions on osteoblast cultures. In vivo studies will determine if systemic PTH administration alters the IGF-IGFBP-IGF protease axis in bone and if deletion of MMP-13 affects PTH and IGF-mediated new bone formation. Together, these studies will provide new insights into mechanisms controlling IGF bioavailability in bone, and how up-stream mediators of bone formation, such as PTH, may exert their affects on osteoblastogenesis through regulating IGF action.

描述（由申请人提供）：胰岛素样生长因子（IGF）是成骨细胞生长和分化的重要介体；但是，六种高亲和力IGF结合蛋白（IGFBPS 1-6）拮抗它们与细胞表面型IGF受体的结合。骨细胞产生的蛋白酶可以将IGFBP降解为低亲和力片段，从而使IGF-IGF受体相互作用发生。我们已经将妊娠相关的血浆蛋白A（PAPP-A）确定为IGFBP-4蛋白酶，MMP-13（胶原酶3）是IGFBP-5蛋白酶，两者都是由成骨细胞系MC3T3-E1产生的。因此，这些IGFBP的蛋白水解处理可能会在成骨细胞生成期间调节IGF作用。我们已经创建了对蛋白水解具有抗性的IGFBP-4和IGFBP-5的重组突变形式，并使用这些构建体来建立稳定转染的MC3T3-E1细胞系，过表达抗蛋白酶的IGFBP突变体。在本文中，我们建议使用这些试剂，以探讨IGFBP蛋白水解如何影响成骨细胞的生长和分化。我们将比较与可切除形式相比，IGFBP-4或-5的蛋白酶形式如何影响成骨细胞生成，以及抑制或下调IGFBP-降解蛋白酶（即PAPP-A和MMP-13）会影响这一过程。在体内，我们将研究IGFBP-4或-5的抗蛋白酶耐蛋白酶或敏感形式如何改变新的骨形成，并且PAPP-A是否可以克服iGFBP-4转基因过量产生对新骨形成的骨基抑制作用。为了探索MMP-13如何介导PTH对骨细胞的合成代谢影响，我们将确定PTH介导的MMP-13上调的钝化可能如何改变体外成骨细胞中IGF作用。我们还将探讨MMP-13 IGFBP-5的抗性形式如何影响PTH的作用对成骨细胞培养物。体内研究将确定全身性PTH给药是否会改变骨中的IGF-IGFBP-IGF蛋白酶轴，以及MMP-13的缺失是否会影响PTH和IGF介导的新骨形成。总之，这些研究将提供有关控制骨骼IGF生物利用度的机制的新见解，以及诸如PTH之类的骨形成的上流介质如何通过调节IGF作用来对成骨细胞生成产生影响。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The role of vitamin D in the metabolic homeostasis of diabetic bone.

维生素 D 在糖尿病骨代谢稳态中的作用。

DOI：
10.1007/s12018-012-9127-9
发表时间：
2013
期刊：
Clinical reviews in bone and mineral metabolism
影响因子：
1.8
作者：
Thrailkill,KathrynM;Fowlkes,JohnL
通讯作者：
Fowlkes,JohnL

Effects of long-term doxycycline on bone quality and strength in diabetic male DBA/2J mice.

DOI：
10.1016/j.bonr.2014.10.001
发表时间：
2015-01
期刊：
Bone reports
影响因子：
2.5
作者：
通讯作者：

Osteo-promoting effects of insulin-like growth factor I (IGF-I) in a mouse model of type 1 diabetes.

DOI：
10.1016/j.bone.2013.07.017
发表时间：
2013-11
期刊：
BONE
影响因子：
4.1
作者：
Fowlkes, John L.;Nyman, Jeffry S.;Bunn, R. Clay;Jot, Chanhee;Wahl, Elizabeth C.;Liu, Lichu;Cockrell, Gael E.;Morris, Lindsey M.;Lumpkin, Charles K., Jr.;Thrailkill, Kathryn M.
通讯作者：
Thrailkill, Kathryn M.

SGLT2 inhibitor therapy improves blood glucose but does not prevent diabetic bone disease in diabetic DBA/2J male mice.

SGLT2 抑制剂治疗可改善糖尿病 DBA/2J 雄性小鼠的血糖，但不能预防糖尿病性骨病。

DOI：
10.1016/j.bone.2015.07.025
发表时间：
2016
期刊：
Bone
影响因子：
4.1
作者：
Thrailkill,KathrynM;ClayBunn,R;Nyman,JeffryS;Rettiganti,MallikarjunaR;Cockrell,GaelE;Wahl,ElizabethC;Uppuganti,Sasidhar;LumpkinJr,CharlesK;Fowlkes,JohnL
通讯作者：
Fowlkes,JohnL

The impact of SGLT2 inhibitors, compared with insulin, on diabetic bone disease in a mouse model of type 1 diabetes.

DOI：
10.1016/j.bone.2016.10.026
发表时间：
2017-01
期刊：
Bone
影响因子：
4.1
作者：
Thrailkill KM;Nyman JS;Bunn RC;Uppuganti S;Thompson KL;Lumpkin CK Jr;Kalaitzoglou E;Fowlkes JL
通讯作者：
Fowlkes JL