NF Center: from animal models to therapeutics

描述(由申请人提供)： 在这份题为：从动物模型到治疗学的申请中，我们请求资金继续支持核因子中心。我们在之前的成功基础上，利用动物模型获得了对肿瘤病因学的新见解，并揭示了肥大细胞作为治疗靶点的作用，目前正处于临床试验中。该中心支持一系列高度互动的多学科研究，旨在(1)阐明NF1中最常见的两种肿瘤：真皮和丛状神经纤维瘤的机制，以及(2)解剖神经纤维蛋白在调节成人内皮和血管平滑肌功能方面的未知功能，这两种细胞系对于神经纤维瘤中肿瘤血管的建立和NF1血管病变的发展至关重要。该中心由一个小型行政核心、一个转基因/动物核心和四个项目组成。转基因/动物核心(PI，Luis F.Parada)负责培育转基因和基因敲除小鼠，基因分型，以及开发或引进更多的小鼠品系。项目1(Pi，Luis F.Parada)是研究NF1突变导致丛状神经纤维瘤和恶性周围神经鞘瘤的分子和细胞机制的富有成效的努力的继续。项目2(PI，D.Wade Clapp)专注于促进神经纤维瘤发生过程中肿瘤微环境中NF1-/-Schwann细胞和不同杂合细胞系之间相互作用的信号机制，并利用FDA批准的治疗药物瞄准这些细胞间的相互作用。项目2还将利用从NF1患者中分离的人类细胞来验证小鼠模型中的观察结果是否在人类系统中真实重现，作为临床前平台，以确定神经纤维瘤微环境中的治疗靶点。项目3(PI，David Ingram)将研究NF1在控制血管内皮细胞和血管平滑肌/周细胞功能中的作用。项目4(Pi，Lu Q.Le)将研究小鼠皮肤神经纤维瘤的病因，并提议对人类患者进行临床试验。项目4还提议建立一个综合性的NF1诊所。总之，拟议的研究计划有望在分子、细胞和系统水平上更好地理解NF1相关的肿瘤发生。 公共卫生相关性：1型神经纤维瘤病是一种不治之症，发病率为每3500名活产儿中就有一名。NF1的显著特征包括咖啡馆Au lait斑点、腋窝和腹股沟雀斑，以及包括真皮和丛状神经纤维瘤在内的多发性周围和中枢神经肿瘤。核因子中心研究人员目前的研究将有助于更好地理解NF1中发生的潜在分子和细胞事件。 2P50NS052606-06/路易斯帕拉达项目1 描述(由申请人提供)： 1型神经纤维瘤病是一种遗传性疾病，对患者具有广泛的后果，从潜在的智力和认知缺陷到出现在周围神经系统的特发性肿瘤，统称为神经纤维瘤。在过去的12年里，我们已经建立了NF1的小鼠模型，目的是总结患者的各种病理特征。这一建议继续并扩展了我们在生成NF1相关神经纤维瘤的忠实基因复制方面的经验。在该奖项资助的之前的工作中，我们的科学团队联手测试了雪旺细胞系外的NF1单倍体功能不全对丛状神经纤维瘤的发展做出了关键贡献的假设。这一假说来自对我们的小鼠模型的研究，结果揭示了肥大细胞在促进肿瘤表型方面的重要性，并最终导致了阻止患者肥大细胞活性的临床试验。在目前的应用中，我们建议扩展我们的小鼠建模能力，以进一步了解丛状神经纤维瘤的病因，识别真皮神经纤维瘤的来源和病因，并使用我们的MPNST模型来寻找治疗机会。在具体目标1中，我们将使用他莫昔芬可诱导的ERE驱动系和替代方法来更好地确定丛状神经纤维瘤的起源细胞来源，并确定肿瘤发展的时间窗口。在具体目标2中，我们将详述我们最近开发的皮肤神经纤维瘤小鼠模型，以及皮肤来源前体(SKP)是这些肿瘤的起源细胞这一发现。我们将使用多种技术，包括使用鸡/鹌鹑胚胎移植神经管来检查神经脊是否是这些肿瘤活性细胞的原始来源。我们还将开发新的有价值的他莫昔芬诱导的转基因Cre驱动系，以探索神经脊源性组织的肿瘤潜能。最后，在特定的目标3中，我们将对MPNST来源的肿瘤细胞进行原代筛选，以进行小分子化学和RNAi高通量筛选。这些筛选旨在识别肿瘤细胞增殖和生长所需的小分子化合物和基因，并可能成为治疗的靶点。 公共卫生相关性：神经纤维瘤和MPNSTs是一种无法治愈的癌症，发生在1型神经纤维瘤病中。我们在小鼠身上创造了NF1基因的突变，使我们能够建立许多疾病病理的可靠模型。在这项应用中，我们使用我们的遗传小鼠模型来研究丛状和真皮神经纤维瘤肿瘤，以获得治疗开发的关键信息。我们还提出了寻找治疗MPNSTs的新的潜在治疗分子的策略。

DESCRIPTION (provided by applicant): In this application, entitled: NF: from animal models to therapeutics, we are requesting funds for continued support of the NF Center. We build on our preceding success in exploiting animal models to gain novel insight into tumor etiology and revealing the mast cell as a therapeutic target that is now in clinical trials. The Center supports a series of highly interactive and multidisciplinary studies aimed at (1) delineating the mechanisms underlying the two most common tumors in NF1: dermal and plexiform neurofibromas and (2) dissecting the unknown functions of neurofibromin in regulating adult endothelial and vascular smooth muscle function - two cell lineages critical for establishment of the tumor vasculature in neurofibromas, and in the development of NF1 vasculopathies. The Center is organized into a small Administrative Core, a Transgenic/Animal Core, and four Projects. The Transgenic/Animal Core (PI, Luis F. Parada) is responsible for breeding transgenic and knockout mice, genotyping, and for developing or importing additional mouse strains. Project 1 (PI, Luis F. Parada) is a continuation of a productive effort to study the molecular and cellular mechanisms by which NF1 mutation engenders plexiform neurofibromas and malignant peripheral nerve sheath tumors. Project 2 (PI, D. Wade Clapp) focuses on the signaling mechanisms that promote interplay between Nf1-/- Schwann cells and different heterozygous cell lineages identified in the tumor microenvironment in neurofibroma development and targeting these cell-cell interactions with FDA-approved therapeutics. Project 2 will also utilize human cells isolated from NF1 patients to verify that the observations in the murine model are faithfully recapitulated in the human system as a preclinical platform to identify therapeutic targets within the neurofibroma microenvironment. Project 3 (PI, David Ingram) will study the role of Nf1 in controlling endothelial and vascular smooth muscle/pericyte function. Project 4 (PI, Lu Q. Le) will investigate the etiology of dermal neurofibromas in mouse and proposes a clinical trial for treatment of human patients. Project 4 also proposes the establishment of a comprehensive NF1 clinic. Together, the proposed program of research promises to contribute to a better understanding of NF1-associated tumorigenesis at the molecular, cellular, and systems levels. PUBLIC HEALTH RELEVANCE: Neurofibromatosis type 1 is an incurable disease with an incidence of 1 in 3,500 live births. Hallmark features of NF1 include cafe au lait macules, and axillary and groin freckling, along with multiple peripheral and central nerve tumors including dermal and plexiform neurofibromas. The current research of the NF Center investigators will lead to a better understanding of the underlying molecular and cellular events that occur in NF1. 2P50NS052606-06/Project 1 PARADA, LUIS DESCRIPTION (provided by applicant): Neurofibromatosis type 1 is a genetic disease with wide ranging consequences on the afflicted individuals ranging from potential intellectual and cognitive deficits to appearance of idiopathic tumors in the peripheral nervous system collectively called neurofibromas. Over the past 12 years we have developed mouse models of NF1 with the objective of recapitulating a variety of the pathologic features seen in patients. This proposal continues and expands upon our experience in generating faithful genocopies of NF1-associated neurofibromas. In preceding work funded by this award, our scientific teams joined forces to test the hypothesis that NF1 haploinsufficiency outside the Schwann cell lineage provided critical contribution to plexiform neurofibroma development. This hypothesis emerged from studies with our mouse models and the outcome has revealed the importance of mast cells in contributing to the tumor phenotype and ultimately leading to clinical trials to block mast cell activity in patients. In the present application we propose to extend our mouse modeling capabilities to further understand the etiology of plexiform neurofibromas, to identify the source and etiology of dermal neurofibromas, and to use our MPNST models to seek out therapeutic opportunities. In Specific Aim 1, we will employ tamoxifen-inducible ere driver lines and alternative approaches to better define the source of the cell of origin for plexiform neurofibromas and to define the temporal window of competence for tumor development. In Specific Aim 2, we will expand on our recent development of a murine model for dermal neurofibromas and on the discovery that skin-derived precursors (SKPs) are the cell of origin for these tumors. We will use multiple techniques including the use of chick/quail embryo transplantation of neural tubes to examine whether the neural crest is the original source of these tumor-competent cells. We will also develop new valuable tamoxifen-inducible transgenic Cre driver lines to probe the neural crest-derived tissues for tumor potential. Finally, in Specific Aim 3, we will screen primary MPNST-derived tumor cells to undertake small chemical and RNAi high throughput screens. These screens aim to identify small molecule compounds and genes that are required for tumor cell proliferation and growth and that can become targets for therapeutics. PUBLIC HEALTH RELEVANCE: Neurofibromas and MPNSTs are incurable cancers that arise in Neurofibromatosis Type 1. We have created mutations in the NF1 gene in mice that allow us to generate faithful models of many of the disease pathologies. In this application, we use our genetic mouse models to study plexiform and dermal neurofibroma tumors to derive critical information for therapy development. We also propose strategies to discover novel potential therapeutic molecules for the treatment of MPNSTs.