MVNP, p62P392L and IL-6 in the Pathogenesis of PD

空间 提供了 麻疹病毒核衣壳基因（MVNP）和p62 p392 L在PD中的作用刚刚开始被研究。 明确了然而，它们如何相互作用导致PD以及涉及的细胞和分子机制 还不清楚我们的假设是，环境因素（MVNP）和遗传因素（p62 P392 L） 有助于PD的发展，MVNP的许多作用是通过其 在OCL谱系的细胞中诱导高水平的IL-6。项目2将研究细胞和体内 MVNP和p62 p392 L共表达的影响，以及参与PD发展的机制。 项目1的研究与项目2密切相关，将研究相关的分子机制 MVNP和p62 PS 92 L的作用导致PD OCL的形成和IL-6的产生增加。到 测试这个假设，我们将：1。确定MVNP和p62 P394 L的共表达是否诱导小鼠中的佩吉特病。 小鼠我们将MVNP基因靶向OCL谱系的小鼠（TRAP-MVNP小鼠）与 p62 P394 L KI（p62 KI）小鼠。我们将确定是否在体外形成具有PD OCL特征的OCL， 根据OCL和成骨细胞活性，OCL形态， 组织形态计量学和pagetic骨病变的发展和程度。的分子基础 在小鼠中MVNP和p62 p394 L共表达时发生的OCL和成骨细胞活性的变化将被 分别在项目1和项目3中描述。2.确定MVNP是否诱导IL-6表达增加 在TRAP-MVNP小鼠中，OCL中的Pagetic OCL和骨病变的发展是负责的。陷阱- MVNP将与IL-6+小鼠交配以产生TRAP-MVNP/IL-B+小鼠。OCL与成骨细胞 这些小鼠的表型，以及它们发生PD特征性局灶性骨病变的能力，将被 与WT、TRAP-MVNP和IL-6+小鼠相比。作为这些实验的一部分，我们将确定 IL-6活性与IL-6中和抗体抑制骨髓培养中pagetic OCL的发展 帕金森病患者。3.检验IL-6表达增加和IL-6受体的存在是否与IL-6受体的表达有关的假设。 p62 P394 L突变可导致小鼠骨质增生和OCL。我们先前已经表明 p62 KI小鼠不发生pagetic骨病变，并且这些小鼠的OCL不是高表达的。 多核或对1，25-（OH）2D 3高度反应，并且不产生升高水平的IL-6，特征 PD OCL的特征。因此，我们将产生TRAP-IL-6小鼠，其中IL-6的表达被抑制。 靶向OCL谱系的细胞，并将它们与p62 KI小鼠杂交。TRAP-IL-6/p62 KI小鼠可在体外培养， 在体外和体内表征其骨表型和OCL和成骨细胞活性，以及 IL-6表达增加和p62 P392 L突变对OCL中NF κ B和p38 MAPK信号传导的影响 和基质细胞。作为这些实验的一部分，我们将治疗正常的人类OCL前体 用含有不同浓度IL-6的p62 P392 L转染，以确定它们是否形成表达IL-6的OCL。 完整的pagetic表型。

SPACE PROVIDED. The role of the measles virus nucleocapsid gene (MVNP) and p62p392L in PD are just beginning to be clarified. However, how they may interact to cause PD and the cellular and molecular mechanisms involved are still unclear. It is our hypothesis that both an environmental factor (MVNP) and a genetic factor (p62P392L) contribute to the development of PD, and that many of the effects of MVNP are mediated through its induction of high levels of IL-6 in cells of the OCL lineage. Project 2 will examine the cellular and in vivo effects of co-expression of MVNP and p62p392L, and the mechanisms involved in the development of PD. Studies in Project 1, which are closely linked to Project 2, will examine the molecular mechanisms involved n the effects of MVNP and p62PS92L that result in formation of PD OCL and increased IL-6 production. To test this hypothesis, we will: 1. Determine if co-expression of MVNP and p62P394L induce Paget's disease in mice. We will mate mice in which the MVNP gene is targeted to the OCL lineage (TRAP-MVNP mice) with p62P394L Kl (p62KI) mice. We will determine if OCLs with the characteristics of PD OCLs form in vitro, and assess the bone phenotype of these mice in terms of OCL and osteoblast activity, OCL morphology, histomorphometry and the development and extent of pagetic bone lesions. The molecular basis for the changes in OCL and osteoblast activity that occur upon co-expression of MVNP and p62p394L in mice will be characterized in Projects 1 and 3 respectively. 2. Determine if increased IL-6 expression induced by MVNP in OCL is responsible for the development of pagetic OCLs and bone lesions in TRAP-MVNP mice. TRAP- MVNP will be mated with IL-6'A mice to generate TRAP-MVNP/IL-B"'' mice. The OCL and osteoblast phenotype of these mice, as well as their capacity to develop focal bone lesions characteristic of PD, will be compared to WT, TRAP-MVNP and IL-6J" mice. As part of these experiments, we will determine if blocking IL-6 activity with a neutralizing antibody to IL-6 inhibits the development of pagetic OCLs in marrow cultures from patients with PD. 3. Test the hypothesis that increased expression of IL-6 and the presence of the p62P394L mutation are sufficient to induce pagetic bone lesions and OCL in mice. We have previously shown that the p62KI mice do not develop pagetic bone lesions, and that OCL from these mice are not hyper- multinucleated or hyper-responsive to 1,25-(OH)2D3, and do not produce elevated levels of IL-6, features characteristic of PD OCLs. Therefore, we will generate TRAP-IL-6 mice in which expression of IL-6 is targeted to cells of the OCL lineage, and interbreed them to the p62KI mice. The TRAP-IL-6/p62KI mice wil be characterized with regard to their bone phenotype and OCL and osteoblast activity in vitro and vivo, and the effects of increased IL-6 expression and p62P392L mutation on NFicB and p38 MAPK signaling in OCLs and stromal cells from these mice. As part of these experiments, we will treat normal human OCL precursors transfected with p62P392L with varying concentrations of IL-6 to determine if they form OCLs that express a complete pagetic phenotype.