NF Center: from animal models to therapeutics

描述（由申请人提供）： 在这份题为“NF：从动物模型到治疗学”的申请中，我们请求为NF中心的持续支持提供资金。我们建立在我们先前的成功，利用动物模型，以获得新的洞察肿瘤病因和揭示肥大细胞作为一个治疗目标，现在在临床试验。该中心支持一系列高度互动和多学科的研究，旨在（1）描绘NF 1中两种最常见肿瘤的机制：真皮和丛状神经纤维瘤和（2）解剖神经纤维蛋白在调节成人内皮和血管平滑肌功能中的未知功能-两种对神经纤维瘤中肿瘤脉管系统的建立至关重要的细胞谱系，和NF 1血管病变的发展。该中心分为一个小的行政核心，一个转基因/动物核心，和四个项目。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，以及开发或进口其他小鼠品系。项目1（PI，Luis F. Parada）是研究NF 1突变产生丛状神经纤维瘤和恶性周围神经鞘肿瘤的分子和细胞机制的富有成效的努力的延续。项目2（PI，D。Wade Clapp）专注于促进Nf 1-/- Schwann细胞与神经纤维瘤发展中肿瘤微环境中鉴定的不同杂合细胞谱系之间相互作用的信号传导机制，并利用FDA批准的疗法靶向这些细胞间相互作用。项目2还将利用从NF 1患者中分离的人类细胞来验证小鼠模型中的观察结果在人类系统中忠实地再现，作为临床前平台来识别神经纤维瘤微环境中的治疗靶点。项目3（PI，大卫英格拉姆）将研究NF 1在控制内皮和血管平滑肌/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，并提出一项治疗人类患者的临床试验。项目4还提议建立一个综合性的NF 1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NF 1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1/3500活产婴儿。NF 1的标志性特征包括咖啡Au lait斑，腋窝和腹股沟雀斑，沿着多发性外周和中枢神经肿瘤，包括真皮和丛状神经纤维瘤。NF中心的研究人员目前的研究将导致更好地了解NF 1中发生的潜在分子和细胞事件。 2 P50 NS 052606 -06/项目1 PARADA，LUIS 描述（由申请人提供）： 1型神经纤维瘤病是一种遗传性疾病，对患病个体具有广泛的后果，从潜在的智力和认知缺陷到外周神经系统中特发性肿瘤的出现，统称为神经纤维瘤。在过去的12年里，我们已经开发了NF 1的小鼠模型，目的是重现患者中观察到的各种病理特征。该建议继续并扩展了我们在产生NF 1相关神经纤维瘤的忠实基因组拷贝方面的经验。在该奖项资助的先前工作中，我们的科学团队联合起来检验了一个假设，即雪旺细胞谱系外的NF 1单倍不足对丛状神经纤维瘤的发展做出了关键贡献。这一假设来自我们的小鼠模型研究，结果揭示了肥大细胞在促成肿瘤表型方面的重要性，并最终导致临床试验以阻断患者中肥大细胞的活性。在本申请中，我们提出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，以确定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，我们将采用他莫昔芬诱导的ere驱动线和替代方法，以更好地确定丛状神经纤维瘤起源细胞的来源，并确定肿瘤发展的时间窗口。在具体目标2中，我们将扩大我们最近开发的皮肤神经纤维瘤小鼠模型，并发现皮肤来源的前体细胞（SKPs）是这些肿瘤的起源细胞。我们将使用多种技术，包括使用鸡/鹌鹑胚胎移植的神经管，以检查是否神经嵴是这些肿瘤细胞的原始来源。我们还将开发新的有价值的他莫昔芬诱导型转基因Cre驱动系，以探测神经嵴衍生组织的肿瘤潜力。最后，在特定目标3中，我们将筛选原发性MPNST衍生的肿瘤细胞，以进行小化学和RNAi高通量筛选。这些筛选旨在鉴定肿瘤细胞增殖和生长所需的小分子化合物和基因，这些化合物和基因可以成为治疗的靶点。 公共卫生相关性：神经纤维瘤和MPNST是1型神经纤维瘤病中出现的不可治愈的癌症。我们已经在小鼠的NF 1基因中创建了突变，使我们能够生成许多疾病病理的忠实模型。在本申请中，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。我们还提出了策略，以发现新的潜在的治疗MPNST的治疗分子。

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.