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

空间 但前提是。 麻疹病毒核衣壳基因(MVNP)和p62p392L在帕金森病中的作用才刚刚开始 澄清了。然而，它们如何相互作用导致帕金森病以及涉及的细胞和分子机制 目前仍不清楚。我们的假设是环境因素(MVNP)和遗传因素(P62P392L)都是 MVNP的许多作用是通过其介导的 诱导OCL系细胞产生高水平的IL-6。项目2将检查细胞和体内的 MVNP和p62p392L共表达的作用及参与帕金森病发生发展的机制。 项目1中与项目2密切相关的研究将检查所涉及的分子机制 N MVNP和p62PS92L对PD OCL形成和IL-6产生的影响。至 验证这一假说，我们将：1.确定MVNP和p62P394L的共同表达是否会诱发Paget病 老鼠。我们将MVNP基因针对OCL谱系的小鼠(TRAP-MVNP小鼠)与 P62P394LK1(P62KI)小鼠。我们将确定具有PD OCLs特征的OCLs是否在体外形成，以及 从OCL和成骨细胞活性、OCL形态和成骨细胞活性等方面评价这些小鼠的骨表型 组织形态计量学与寻常型骨病损的发展和程度。基因的分子基础 MVNP和p62p394L共表达对小鼠OCL和成骨细胞活性的影响 分别在项目1和项目3中描述了其特点。2.确定MVNP是否诱导IL-6表达增加 在OCL中负责在TRAP-MVNP小鼠中发生寻常型OCL和骨损伤。陷阱- MVNP将与IL-6‘A小鼠交配，产生TRAP-MVNP/IL-B“’小鼠。OCL和成骨细胞 这些小鼠的表型以及它们发展为帕金森病特有的局灶性骨骼病变的能力将是 与WT、TRAP-MVNP和IL-6J小鼠相比。作为这些实验的一部分，我们将确定是否阻断 具有IL-6中和抗体的IL-6活性抑制骨髓培养中传播性OCL的发展 来自帕金森氏症患者。3.检验以下假设：IL-6表达增加和 P62P394L突变足以诱发小鼠变应性骨损伤和OCL。我们之前已经展示了 P62KI小鼠不会出现寻常型骨质损害，这些小鼠的OCL也不会过度敏感。 多核或对1，25-(OH)2D3高度反应，不产生升高的IL-6水平，特征 PD OCL的特点。因此，我们将产生表达IL-6的TRAP-IL-6小鼠 靶向OCL血统的细胞，并将它们与p62KI小鼠杂交。TRAP-IL-6/p62KI小鼠 在体外和体内研究它们的骨表型、OCL和成骨细胞活性 IL-6表达增加和p62P392L突变对OCL中NFicB和p38MAPK信号的影响 以及这些小鼠身上的基质细胞。作为这些实验的一部分，我们将治疗正常的人类OCL前体 用不同浓度的IL-6转染p62P392L，以确定它们是否形成表达a的OCL 完整的传教士表型。

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.