Identification of Specific Antagonists that Bind Hedgehog and Block its Activity
鉴定结合 Hedgehog 并阻断其活性的特定拮抗剂
基本信息
- 批准号:8100944
- 负责人:
- 金额:$ 36.29万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2011
- 资助国家:美国
- 起止时间:2011-03-01 至 2015-02-28
- 项目状态:已结题
- 来源:
- 关键词:AffectAffinityAffinity ChromatographyApoptosisBindingBinding SitesBiological AssayBiological FactorsCancer ModelCancer cell lineCellsChemicalsClinicCollectionComplexDNADNA SequenceDevelopmentDiffusionDiseaseDoseDrug resistanceErinaceidaeExtracellular ProteinFDA approvedFamilyFluorescence PolarizationFutureGenesGoalsGrowthHeparan Sulfate ProteoglycanHeparinHeparin AntagonistsHeparin BindingHeparitin SulfateHumanInorganic SulfatesLengthLibrariesLigandsLightLiverLungMalignant NeoplasmsMalignant neoplasm of prostateMapsMeasuresMediatingOligonucleotidesPancreasPathway interactionsPatternPeptide aptamersPeptidesPhasePlayProteinsResearchResistanceRoleScreening procedureSeriesSignal TransductionSignaling ProteinSiteSolidStagingSuraminTestingTissuesUnspecified or Sulfate Ion SulfatesWestern BlottingWorkanti-cancer therapeuticaptamerbasebiochipcancer cellcancer typecounterscreeneffective therapyheparin proteoglycanhuman SMO proteinin vivo Modelinhibitor/antagonistinterestmigrationnovelnovel strategiesnovel therapeuticspeptide analogperlecanprotein protein interactionreceptorresponsesmall moleculesmall molecule librariessmoothened signaling pathwaysynthetic peptidetherapeutic targettooltraffickingtranscription factortumor
项目摘要
DESCRIPTION (provided by applicant): The long term goal of this initiative is to identify novel antagonists that target hedgehog (Hh) binding interactions important for Hh activity as the basis for the development of novel anti-cancer therapeutics. Aberrant expression of the Hh pathway has been implicated in the growth of approximately 20-25% of all cancers, with many driven by the over-expression of Hh ligand. Several natural product and chemical inhibitors that block Hh signaling have anti-tumor effects in models of cancer. These compounds work predominantly by targeting the Hh co-receptor Smoothened (Smo) or Gli transcription factor. Resistance to Smo inhibitors has been observed in the clinic and hence inhibitors targeting other components of the pathway are greatly needed. There is compelling evidence that heparin sulfate proteoglycans (HSPGs) are critically involved in Hh ligand trafficking and signaling activity with a role for HSPGs implicated by restriction in the range of Hh signaling in cells lacking the function of the tout velu gene, which encodes a heparin sulfate copolymerase. The goal of this proposal is to identify novel inhibitors of the Hh pathway by targeting the interaction of Hh ligand with HSPGs. We propose to do this using a novel Hh/heparin binding assay we have developed to identify molecules that can modulate heparin-Hh interactions. In this assay, unlabeled heparin molecules and the small molecule suramin inhibit this interaction and also blocked Hh activity in a cell based assay of Hh function. This assay will be used in high throughput mode to screen a library of 60,000 small molecules to identify small molecule antagonists of Hh/heparin binding. Small molecule mediated disruption of protein- protein interactions involving extracellular proteins such as Hh and their binding partners is challenging and we expect aptamer selection to provide an alternative and complementary approach to identifying antagonists of Hh protein mediated activity whether at the heparin binding site on Hh or elsewhere on the protein. Hence, the goals of the current proposal are to use the Hh/heparin binding assay as a primary screen to identify compounds from large chemical libraries that block Hh binding to heparin and to map the heparin binding site of Hh using synthetic peptides and (Specific Aim 1). To isolate and characterize Hh binding aptamers (Specific Aim 2). Use a series of secondary/orthogonal assays to first confirm and verify hits from primary screening and aptamers from the selection, and secondly identify those that selectively block Hh/HSPG association and inhibit Hh signaling in bioassays and affect proliferation, survival and migration of Hh- dependent cancer cell lines (Specific Aim 3). The various secondary and counter screens will be employed to further characterize the selectivity and activity of the identified molecules, setting the stage for future chemical optimization and subsequent in vivo models of cancer. We believe the resulting molecules will be especially valuable as research tools to better understand HSPG contributions to Hh activity and as novel inhibitors to probe diseases associated with Hh-ligand driven cancers.
PUBLIC HEALTH RELEVANCE: The Hedgehog (Hh) pathway is a compelling therapeutic target as it plays a central role in the growth of a vast array of human cancer types including those such as pancreatic, liver and lung with few effective treatment options. The few current inhibitors identified to date for the Hh pathway almost exclusively bind to the smoothened co-receptor or target the Gli transcription factors and hence inhibitors targeting the Hh pathway by alternative mechanisms are of great interest. There is accumulating evidence that the ability of Hh protein to function is modulated by heparin sulfate proteoglycans (HSPG), and so we propose to identify small molecule and DNA-based compounds that selectively block Hh/HSPG interactions, test them as inhibitors in cancers models and so potentially identify a novel therapeutic avenue for hedgehog- dependent cancers.
描述(由申请人提供):该计划的长期目标是鉴定靶向对Hh活性重要的Hedgehog(Hh)结合相互作用的新型拮抗剂,作为开发新型抗癌治疗剂的基础。Hh途径的异常表达与大约20-25%的所有癌症的生长有关,其中许多是由Hh配体的过表达驱动的。阻断Hh信号传导的几种天然产物和化学抑制剂在癌症模型中具有抗肿瘤作用。这些化合物主要通过靶向Hh共受体Smoothened(Smo)或Gli转录因子起作用。在临床中已经观察到对Smo抑制剂的抗性,因此非常需要靶向该途径的其他组分的抑制剂。有令人信服的证据表明,硫酸肝素蛋白聚糖(HSPG)是关键参与Hh配体运输和信号传导活性的作用,HSPG的牵连限制在细胞中的Hh信号传导的范围内缺乏tout velu基因的功能,它编码的硫酸肝素共聚酶。该提案的目标是通过靶向Hh配体与HSPG的相互作用来鉴定Hh通路的新型抑制剂。我们建议这样做,我们已经开发了一种新的Hh/肝素结合试验,以确定分子,可以调节肝素Hh相互作用。在该测定中,未标记的肝素分子和小分子苏拉明抑制这种相互作用,并且还在基于细胞的Hh功能测定中阻断Hh活性。该测定将以高通量模式用于筛选60,000个小分子的文库,以鉴定Hh/肝素结合的小分子拮抗剂。小分子介导的蛋白质-蛋白质相互作用的破坏涉及细胞外蛋白质如Hh及其结合配偶体是具有挑战性的,我们期望适体选择提供一种替代和互补的方法来鉴定Hh蛋白质介导的活性的拮抗剂,无论是在Hh上的肝素结合位点还是在蛋白质上的其他地方。因此,当前提案的目标是使用Hh/肝素结合试验作为初步筛选,以从大型化学文库中鉴定阻断Hh与肝素结合的化合物,并使用合成肽和(具体目标1)绘制Hh的肝素结合位点。分离和表征Hh结合适体(具体目标2)。使用一系列二次/正交试验,首先确认和验证来自初步筛选的命中物和来自选择的适体,其次鉴定在生物试验中选择性阻断Hh/HSPG缔合和抑制Hh信号传导并影响Hh依赖性癌细胞系的增殖、存活和迁移的那些适体(具体目标3)。将采用各种次级和反向筛选来进一步表征所鉴定分子的选择性和活性,为未来的化学优化和随后的癌症体内模型奠定基础。我们相信,所得到的分子将是特别有价值的研究工具,以更好地了解HSPG的贡献Hh活性和作为新的抑制剂,探针与Hh-配体驱动的癌症相关的疾病。
公共卫生关系:Hedgehog(Hh)通路是一个引人注目的治疗靶点,因为它在包括胰腺癌、肝癌和肺癌在内的多种人类癌症类型的生长中起着核心作用,而有效的治疗方案却很少。迄今为止鉴定的用于Hh途径的几种当前抑制剂几乎专门结合平滑的共受体或靶向Gli转录因子,因此通过替代机制靶向Hh途径的抑制剂引起了极大的兴趣。越来越多的证据表明,Hh蛋白发挥功能的能力受到硫酸肝素蛋白聚糖(HSPG)的调节,因此我们提出鉴定选择性阻断Hh/HSPG相互作用的小分子和基于DNA的化合物,在癌症模型中测试它们作为抑制剂,从而潜在地鉴定刺猬依赖性癌症的新治疗途径。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Kevin Peter Williams其他文献
Kevin Peter Williams的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Kevin Peter Williams', 18)}}的其他基金
Prenatal Alcohol Exposure and Fetal Alcohol Spectrum Disorder
产前酒精暴露和胎儿酒精谱系障碍
- 批准号:
10541715 - 财政年份:2022
- 资助金额:
$ 36.29万 - 项目类别:
Prenatal Alcohol Exposure and Fetal Alcohol Spectrum Disorder
产前酒精暴露和胎儿酒精谱系障碍
- 批准号:
10705759 - 财政年份:2022
- 资助金额:
$ 36.29万 - 项目类别:
Project 2: Prenatal Alcohol Exposure (PAE) and Fetal Alcohol Spectrum Disorder (FASD)
项目2:产前酒精暴露(PAE)和胎儿酒精谱系障碍(FASD)
- 批准号:
10540966 - 财政年份:2022
- 资助金额:
$ 36.29万 - 项目类别:
Project 2: Prenatal Alcohol Exposure (PAE) and Fetal Alcohol Spectrum Disorder (FASD)
项目2:产前酒精暴露(PAE)和胎儿酒精谱系障碍(FASD)
- 批准号:
10705860 - 财政年份:2022
- 资助金额:
$ 36.29万 - 项目类别:
Immune Microenvironments and Hepatocyte Growth Factor Signaling Interactions in Breast Cancer Disparities
乳腺癌差异中的免疫微环境和肝细胞生长因子信号传导相互作用
- 批准号:
10708080 - 财政年份:2017
- 资助金额:
$ 36.29万 - 项目类别:
Immune Microenvironments and Hepatocyte Growth Factor Signaling Interactions in Breast Cancer Disparities
乳腺癌差异中的免疫微环境和肝细胞生长因子信号传导相互作用
- 批准号:
10556581 - 财政年份:2017
- 资助金额:
$ 36.29万 - 项目类别:
Identification of potent and selective GLI1 inhibitors
有效且选择性 GLI1 抑制剂的鉴定
- 批准号:
8523361 - 财政年份:2013
- 资助金额:
$ 36.29万 - 项目类别:
The Hedgehog Pathway and Inflammatory Breast Cancer
刺猬通路和炎症性乳腺癌
- 批准号:
7666030 - 财政年份:2008
- 资助金额:
$ 36.29万 - 项目类别:
The Hedgehog Pathway and Inflammatory Breast Cancer
刺猬通路和炎症性乳腺癌
- 批准号:
7427272 - 财政年份:2008
- 资助金额:
$ 36.29万 - 项目类别:
The Hedgehog Pathway and Inflammatory Breast Cancer
刺猬通路和炎症性乳腺癌
- 批准号:
7922003 - 财政年份:2008
- 资助金额:
$ 36.29万 - 项目类别:
相似海外基金
Cellular membrane affinity chromatography kit for drug discovery
用于药物发现的细胞膜亲和层析试剂盒
- 批准号:
10506915 - 财政年份:2021
- 资助金额:
$ 36.29万 - 项目类别:
Cellular membrane affinity chromatography kit for drug discovery
用于药物发现的细胞膜亲和层析试剂盒
- 批准号:
10325006 - 财政年份:2021
- 资助金额:
$ 36.29万 - 项目类别:
SBIR Phase I: A New Class of Immobilized Metal Affinity Chromatography Resins
SBIR 第一阶段:一类新型固定金属亲和色谱树脂
- 批准号:
1746198 - 财政年份:2018
- 资助金额:
$ 36.29万 - 项目类别:
Standard Grant
Marine speciation of nickel using immobilized nickel affinity chromatography
使用固定镍亲和色谱法测定镍的海洋形态
- 批准号:
512537-2017 - 财政年份:2017
- 资助金额:
$ 36.29万 - 项目类别:
University Undergraduate Student Research Awards
I-Corps: Commercialization of Immobilized Metal Affinity Chromatography Resins Based on Nanomaterials
I-Corps:基于纳米材料的固定化金属亲和层析树脂的商业化
- 批准号:
1404605 - 财政年份:2014
- 资助金额:
$ 36.29万 - 项目类别:
Standard Grant
Antibody Purification via Affinity Chromatography that Utilizes the Unconventional Nucleotide Binding Site
利用非常规核苷酸结合位点通过亲和色谱法纯化抗体
- 批准号:
1263713 - 财政年份:2013
- 资助金额:
$ 36.29万 - 项目类别:
Continuing Grant
Development of multivalent DNA network based affinity chromatography diagnostics for isolating circulating tumour cells
开发基于多价 DNA 网络的亲和色谱诊断法,用于分离循环肿瘤细胞
- 批准号:
425749-2012 - 财政年份:2012
- 资助金额:
$ 36.29万 - 项目类别:
Postgraduate Scholarships - Master's
Next-Generation Affinity Chromatography with PEGylated Ligands
使用聚乙二醇化配体的新一代亲和色谱法
- 批准号:
1159886 - 财政年份:2012
- 资助金额:
$ 36.29万 - 项目类别:
Standard Grant
Immobilized zirconium ion affinity chromatography for specific enrichment of phosphoproteins
用于磷蛋白特异性富集的固定化锆离子亲和层析
- 批准号:
19560760 - 财政年份:2007
- 资助金额:
$ 36.29万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Accelerating drug discovery using frontal affinity chromatography/mass spectrometry
使用正面亲和色谱/质谱加速药物发现
- 批准号:
234753-2000 - 财政年份:2003
- 资助金额:
$ 36.29万 - 项目类别:
Collaborative Research and Development Grants