Identification of Novel Cancer Selective Compounds in 3D Tumor Organoid Assays

摘要 乳腺癌是一种病理上和基因上不同的疾病，反映了不同的信号通路。 对癌症的发展负有责任。乳腺癌的体外模型必须结合复杂的性质 疾病的培养方法与患者所见的不同临床类型的癌症非常相似。二零零七年， 乳腺癌新增病例超过17.8万例，死亡4万例，成为全球第二大乳腺癌 女性癌症相关死亡的原因，这些统计数据表明需要制定更多的 有效治疗(美国癌症协会；http://www.cancer.org).在过去的十年里，最大的进步 在乳腺癌的化疗治疗中涉及到剂量的优化和时间安排 标准疗法。几种新的靶向疗法和生物制剂已被批准用于治疗 乳腺癌；然而，许多癌症即使用理想的药物方案也不起作用或复发。因为 特定的分子标记可以将乳腺癌分为多个亚型，具有不同的临床特征 特征，我们现在可以针对每个人选择性地追求治疗的发展 肿瘤类型。因此，乳腺癌必须被认为是一种多方面的疾病，每种亚型都具有独特性。 化疗干预的机会。 我们已经开发出一种新的抗癌药物发现屏幕，它包含了细胞和基因 乳腺癌的多样性。这种筛查的独特之处在于它在分子上使用了小鼠原发肿瘤器官 分类为与在患者中观察到的实际乳腺癌亚型相似。亚型特异性肿瘤有机体 根据化学文库进行筛选，以确定新的小分子抗癌化合物。同时， 化合物也被筛选出它们对正常的初级有机物的影响。使用此屏幕，我们确定了一个 新型小分子C-6，可杀死来自小鼠和人类的原发肿瘤组织，以及随后的 构效关系分析揭示了该分子的基本药效团。重要的是，C-6 对从小鼠和小鼠体内分离的基底样癌和腔癌有机化合物具有选择性细胞毒作用 而不是从正常小鼠和人体组织中分离出来的有机化合物。这个化合物的结构是 新奇的，可能通过独特的作用机制杀死肿瘤细胞。我们观察到C-6延迟上皮细胞 正常乳腺器官腔内的细胞凋亡；然而，它不影响分支 乳腺器官的形态发生。C-6的选择性细胞毒作用及其对细胞毒性的影响 正常的生物学过程表明，这种化合物针对的是对体内平衡至关重要的分子途径。 乳腺癌细胞。因此，本提案中概述的目标将建立最佳的药效团 C-6的结构，确定分子靶点(S)，并评价其在肿瘤和正常组织中的生物学作用 细胞。

ABSTRACT Breast cancer is a pathologically and genetically heterogeneous disease, reflecting diverse signaling pathways responsible for cancer progression. In vitro models of breast cancer must integrate the complex nature of the disease with culturing methods that closely mimic different clinical types of cancer seen in patients. In 2007, there were more than 178,000 new cases and 40,000 deaths from breast cancer, making it the second leading cause of cancer related death in women and these statistics suggest the need for the development of more effective treatments (American Cancer Society; http://www.cancer.org). Over the past ten years, most advances in chemotherapeutic treatment of breast cancer have involved the optimization of doses and scheduling of standard therapeutics. Several new targeted therapies and biologics have been approved for the treatment of breast cancer; however, many cancers do not respond to or recur even with the ideal drug regimens. Because specific molecular signatures can classify breast cancer into multiple subtypes with distinct clinical characteristics, we can now pursue the development of therapeutics with selectiveness against each individual tumor type. Thus, breast cancer must be considered a multifaceted disease with each subtype offering unique opportunities for chemotherapeutic intervention. We have developed a novel anti-cancer drug discovery screen that encompasses the cellular and genetic diversity of breast cancer. This screen is unique in that it uses murine primary tumor organoids molecularly classified as similar to actual breast cancer subtypes observed in patients. The subtype-specific tumor organoids are screened against a chemical library to identify novel small-molecule anti-cancer compounds. In parallel, the compounds are also screened for their effect on normal primary organoids. Using this screen, we identified a novel small molecule, C-6, that kills primary tumor tissue derived from both mice and humans, and subsequent structure activity relationship analyses revealed the basic pharmacophore of the molecule. Importantly, C-6 exhibits selective cytotoxicity against both basal-like and luminal cancer organoids isolated from mice and human patients, but not organoids isolated from normal mice and human tissue. This compound is structurally novel and may kill tumor cells through a unique mechanism of action. We observed that C-6 delays epithelial cell apoptosis within the lumens of normal mammary organoids; however, it did not affect branching morphogenesis of mammary organoids. The selective cytotoxicity of C-6 combined with its minimal affect on normal biological processes suggests that this compound targets a molecular pathway vital to the homeostasis of breast cancer cells. Thus, the objectives outlined in this proposal will establish the optimal pharmacophore structure of C-6, identify the molecular target(s), and evaluate its biological effects in both tumor and normal cells.