Defining Melanoma Therapeutic Avenues by Integrative Functional Genomics

通过综合功能基因组学定义黑色素瘤治疗途径

• 批准号: 7431956
• 负责人: Levi A. Garraway
• 金额: $ 256.5万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7431956
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Although tumor classification and patient stratification based on genomic criteria offers tremendous clinical potential, discerning critical effectors of tumor genetic alterations—and developing robust therapeutic avenues to intercept them—represents a formidable obstacle to translational oncology. Recent advances in large-scale `perturbagen' approaches (e.g., viral RNAi and small molecule screening) hold great promise to alleviate such bottlenecks; however, their systematic application in cancer biology would benefit markedly from robust in vitro cancer models that fully encompass the genomic diversity manifest in patients. Malignant melanoma offers a rich avenue in this regard: unlike other solid tumors, cells from this lethal malignancy are readily cultured in vitro, thereby providing a diverse and tractable system for genomic and functional studies. Accordingly, we propose to apply pooled RNAi and small molecule microarray screening sequentially to a panel of genetically characterized and patient-derived melanoma cell lines. We will assemble a lentiviral library targeting all expressed genes located within the portion of the genome that is amplified in melanoma, and perform pooled RNAi screening across a panel of 20 melanoma lines representative of the most prevalent melanoma genomic alterations. `Hits' (e.g., selectively depleted shRNAs) will be correlated with genomic patterns to identify (onco)gene targets of melanoma amplifications. We will validate the most promising target genes using a series of cell survival assays in vitro (in arrayed RNAi format) and tumor formation assays in vivo (using shRNA `mini-pools'). Finally, we will identify candidate ligands that bind the top target (onco)protein candidates by performing small-molecule microarray screens using epitope-tagged protein constructs. If successful, this project should elaborate a spectrum of target proteins and potential lead compounds linked to common genetic changes in melanoma. Moreover, these efforts should inform a “platformizable” integrated approach applicable to all cancers for which tractable in vitro models exist.

尽管肿瘤分类和患者分层基于基因组标准 提供巨大的临床潜力，识别肿瘤遗传的关键效应因子 改变——并开发强大的治疗途径来拦截它们——代表了一种 转化肿瘤学的巨大障碍。大规模的最新进展 “扰动”方法（例如病毒 RNAi 和小分子筛选）效果很好 承诺缓解此类瓶颈；然而，它们在癌症中的系统应用 生物学将显着受益于强大的体外癌症模型，这些模型完全涵盖 基因组多样性体现在患者身上。恶性黑色素瘤提供了丰富的途径 在这方面：与其他实体瘤不同，来自这种致命恶性肿瘤的细胞很容易 体外培养，从而为基因组和 功能研究。 因此，我们建议应用混合 RNAi 和小分子微阵列 按顺序筛选一组具有遗传特征和源自患者的 黑色素瘤细胞系。我们将组装一个针对所有表达基因的慢病毒文库 位于黑色素瘤中扩增的基因组部分内，并执行 对一组 20 个黑色素瘤系进行汇总 RNAi 筛查，这些黑色素瘤系代表了最 普遍的黑色素瘤基因组改变。 “命中”（例如，选择性耗尽的 shRNA）将 与基因组模式相关以识别黑色素瘤的（癌）基因靶点 放大。我们将使用一系列细胞验证最有希望的靶基因 体外存活测定（以阵列 RNAi 形式）和体内肿瘤形成测定 （使用 shRNA“迷你池”）。最后，我们将确定结合顶部的候选配体 通过使用以下方法进行小分子微阵列筛选来靶向（癌）蛋白质候选物 表位标记的蛋白质构建体。如果成功，该项目应该制定一个频谱 与常见遗传变化相关的目标蛋白和潜在先导化合物 黑色素瘤。此外，这些努力应该为“平台化”集成方法提供信息 适用于所有存在可处理体外模型的癌症。

项目成果

Stepping on the GAS: a brake pedal for melanoma metastasis?

踩下GAS：黑色素瘤转移的刹车踏板？

• DOI: 10.1111/j.1755-148x.2008.00533.x
• 发表时间: 2009
• 期刊: Pigment cell & melanoma research
• 影响因子: 4.3
• 作者: Linja,Marika;Garraway,LeviA
• 通讯作者: Garraway,LeviA