The Development of Screening Assays For Novel Inhibitors Of ARNO And Its Effector

描述(申请人提供)：癌症是美国第二大死因，仅次于心脏疾病，约占所有死亡人数的23%。尽管在许多癌症的治疗方面取得了重大进展，但据估计，2010年将有569,490名美国人死于这些疾病(《癌症事实和数字2010》，美国癌症协会)，而且迄今还没有针对多种癌症的有效治疗方法。癌症的分子复杂性使得识别更多的致癌途径成为治疗干预的首要任务。2010年6月，NIH发布了一份计划公告(PA-10-213)，用于开发用于探针和治疗前发现的高通量筛查分析，以鼓励开发新的、具有科学价值的分析，这些分析有可能开发成相关的、高优先级疾病靶点的高通量筛查(HTS)，用于识别小分子工具和候选治疗。我们和我们的合作者共同提出的数据提供了令人信服的证据，证明抑制细胞粘附素，激活Arf家族GTP酶的鸟核苷酸交换因子，在非常不同的细胞谱系中产生抗癌活性。此外，我们证明了这种抑制在RAS/Raf/ERK结构性激活的背景下减少了体外增殖和侵袭。这些受体酪氨酸激酶信号通路下游成分的突变极大地限制了目前可用的癌症治疗的实用性和有效性。因此，我们认为，靶向细胞粘附素Arno及其效应物Arf6可能会产生有希望的新的小分子抑制剂，可用于对抗先天和获得性化疗耐药。在本申请中，我们寻求具体解决NIH定义的需求，并提出了一种策略，以开发符合HTS运动的分析方法，用于识别和评估Arno和/或Arf6负调制子。如果成功，我们将寻求通过与美国国立卫生研究院最近推出的分子图书馆和成像计划或其他机构合作，实现这些筛选的自动化，这些机构可以访问大型、多样化的化合物图书馆。从长远来看，这一策略有望识别新的小分子探针和针对这些新的致癌靶点的治疗前化合物。

DESCRIPTION (provided by applicant): Cancer is the second leading cause of death in the United States-second only to diseases of the heart-and accounts for about 23% of all deaths. Although significant progress has been made in the treatment of many cancers, it is estimated that in 2010, 569,490 Americans will die from these diseases (Cancer Facts and Figures 2010, American Cancer Society) and there is as yet no efficacious treatment for many forms of cancers. The molecular complexity of cancer makes the identification of additional oncogenic pathways amenable to therapeutic intervention paramount. In June 2010, NIH posted a program announcement (PA-10- 213) for the Development of Assay for High-throughput Screening for Use in Probe and Pre-therapeutic Discovery to encourage the development of novel, scientifically outstanding assays that have the potential to be developed into high-throughput screens (HTS) of relevant, high priority disease targets both for the identification of small molecule tools and therapeutic candidates. Together, we and our collaborators present data that provide compelling evidence that the inhibition of cytohesins, the guanine nucleotide exchange factors that activate Arf family GTPases, produces anticancer activity in vastly different cell lineages. Further, we demonstrate that this inhibition reduces in vitro proliferation and invasion in the context of Ras/Raf/ERK constitutive activation. Mutations of these downstream components of receptor tyrosine kinase signaling pathways have greatly limited the utility and effectiveness of currently available cancer therapies. Therefore, we believe targeting the cytohesin, ARNO, and its effector, Arf6, may produce promising new small molecule inhibitors useful at combating both innate and acquired chemotherapeutic resistance. In this application we seek to specifically address the need defined by NIH and present a strategy to developing assays amenable to a HTS campaign intended for the identification and evaluation of ARNO and/or Arf6 negative modulators. If successful, we will then seek to automate these screens through collaborations with the NIH's recently launched Molecular Libraries and Imaging initiative or other institutions with access to large, diverse compound libraries. In the long term, this strategy promises to identify new small molecule probe and pre-therapeutic compounds against these novel oncogenic targets.