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OSTEOBLAST AND PTH/PTHRP BONE RESORPTION

成骨细胞和 PTH/PTHRP 骨吸收

基本信息

批准号：
2376650
负责人：
KARL Leonard INSOGNA
金额：
$ 21.29万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1989
资助国家：
美国
起止时间：
1989-08-01 至 1999-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2376650
关键词：
biological signal transduction colony stimulating factor genetic library homeostasis in situ hybridization laboratory rat molecular biology newborn animals northern blottings nuclear runoff assay nucleic acid sequence osteoblasts parathyroid hormone related protein parathyroid hormones pathologic bone resorption receptor expression

项目摘要

The osteoblast or an osteoblast-like cell is the principal to which parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP) bind in bone. We hypothesize that osteoblast-derived factors are therefore critically important local mediators of PTH-and PTHrP-induced bone resorption. A corollary to this hypothesis is that differences in the actions of PTH and PTHrP on osteoblasts may help explain the different in vivo effects of these two hormones. To pursue this hypothesis, the effects of these two agonists on osteoblasts-derived factors which may 1) activate mature osteoclasts or 2) stimulate recruitment of pre-osteoclasts will be examined. We have observed that PTH and PTHrP-treated osteoblasts release a soluble bone-resorbing activity. This activity has been partially purified and can directly stimulate resorption by isolated osteoclasts (under conditions where PTHrP is inactive). This material will be purified to homogeneity to obtain primary structural information. Amino-acid sequence information will be used to synthesize oligonucleotide probes for screening an Saos-2cDNA library. Based on our previous work, demonstrating PTH-induce GM-CSF release from osteoblasts, this molecule will be studied as an example of an osteoblast-derived factor which may regulate osteoclast recruitment. The molecular mechanisms underlying PTH/PTHrP-stimulated release of GM-CSF from osteoclast will be examining the effects of neutralizing antisera to GM-CSF on PTH/PTHrP-induced resorption in the fetal rat long-bone and mouse metatarsal assays. We have very recently identified CSF-1 receptors (c-fms) in mature rat osteoclasts. Further, in preliminary studies we have identified down- regulation of this receptor by GM-CSF suggesting an important interaction between these two colony-stimulating factors in regulating osteoclast biology. These observations will be pursued using immunocytochemical techniques and in situ hybridization. Specifically, we will study whether the effect of GM-CSF on c-fms occurs at the level of transcription or translation. Finally, a mammalian expression system employing rat insulinoma cells has been developed using the three available cDNA clones for PTHrP. The effects of amino-terminal purified recombinant PTHrP on signal transduction mechanisms in, and release of resorptive cytokines from, osteoblasts will be studied. In the aggregate, these studies should allow a more precise picture of the cellular mechanisms by which PTH and PTHrP induce resorption in bone and should lead to a clearer picture of how these compounds act under physiologic conditions as well as those of disturbed mineral homeostasis.

成骨细胞或成骨细胞样细胞是主要的，甲状旁腺激素（PTH）和甲状旁腺相关蛋白（PTHrP）在骨头上绑起来。我们假设成骨细胞衍生因子是因此，PTH和PTHrP诱导的骨吸收这一假设的一个推论是， PTH和PTHrP对成骨细胞的作用可能有助于解释这两种激素在体内的不同作用。追求这个假设，这两种激动剂对成骨细胞源性的影响可能1）激活成熟破骨细胞或2）刺激将检查前破骨细胞的募集。我们已经观察到PTH和PTHrP处理的成骨细胞释放可溶性骨吸收活性。该活性已被部分纯化，可以直接刺激分离的破骨细胞的吸收（在 PTHrP失活的条件）。这些材料将被纯化，同质性以获得初级结构信息。氨基酸序列信息将用于合成寡核苷酸探针筛选Saos-2cDNA文库。基于我们之前的工作，证明PTH诱导GM-CSF从成骨细胞释放，这种分子将作为成骨细胞衍生因子的一个例子进行研究，调节破骨细胞募集。的分子机制将检查PTH/PTHrP刺激的破骨细胞释放GM-CSF GM-CSF中和抗血清对PTH/PTHrP诱导的在胎鼠长骨和小鼠跖骨测定中的吸收。我们最近在成年大鼠中鉴定了CSF-1受体（c-fms），破骨细胞此外，在初步研究中，我们已经确定下来- GM-CSF对该受体的调节表明了重要的相互作用这两种集落刺激因子在调节破骨细胞中的作用生物学这些观察结果将采用免疫细胞化学技术和原位杂交。具体来说，我们将研究 GM-CSF对c-fms的作用是否发生在转录或翻译。最后，使用大鼠胰岛素瘤细胞的哺乳动物表达系统具有使用PTHrP的三个可用cDNA克隆开发。的氨基端纯化的重组PTHrP对信号的影响再吸收性细胞因子的转导机制和释放，将研究成骨细胞。总体而言，这些研究应允许更精确地了解PTH和 PTHrP诱导骨吸收，并应导致更清晰的图片，这些化合物如何在生理条件下起作用，以及矿物质平衡紊乱

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Treatment of humoral hypercalcemia of malignancy in rats with inhibitors of carbonic anhydrase.

用碳酸酐酶抑制剂治疗大鼠恶性肿瘤体液高钙血症。

DOI：
10.1002/jbmr.5650051007
发表时间：
1990
期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
0
作者：
Brown,GM;Morris,CA;Mitnick,MA;Insogna,KL
通讯作者：
Insogna,KL

Parathyroid hormone-related protein. Evidence for secretion of a novel mid-region fragment by three different cell types.

甲状旁腺激素相关蛋白。

DOI：
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Soifer,NE;Dee,KE;Insogna,KL;Burtis,WJ;Matovcik,LM;Wu,TL;Milstone,LM;Broadus,AE;Philbrick,WM;Stewart,AF
通讯作者：
Stewart,AF

Parathyroid hormone-related protein: evidence for isoform- and tissue-specific posttranslational processing.

甲状旁腺激素相关蛋白：异构体和组织特异性翻译后加工的证据。

DOI：
10.1021/bi00189a054
发表时间：
1994
期刊：
Biochemistry
影响因子：
2.9
作者：
Yang,KH;dePapp,AE;Soifer,NE;Dreyer,BE;Wu,TL;Porter,SE;Bellantoni,M;Burtis,WJ;Insogna,KL;Broadus,AE
通讯作者：
Broadus,AE

Evidence for a functional association between phosphatidylinositol 3-kinase and c-src in the spreading response of osteoclasts to colony-stimulating factor-1.