High-throughput screening for small molecules with specific toxicity for breast c

描述(由申请人提供)：有一个重要的未得到满足的需求，即识别能够靶向乳腺癌干细胞(CSCs)的小分子。肿瘤干细胞负责推动肿瘤的生长和转移。众所周知，CSCs对包括化疗和放射治疗在内的大多数传统疗法也具有抵抗力，这清楚地强调了针对CSCs的治疗的必要性。这项拟议的研究的目的是确定对乳腺CSCs具有选择性毒性的小分子探针。这些分子将对探索干细胞的生物学非常有用，也将成为有前途的治疗药物候选者。我们建议使用我们开发的一种新方法来筛选靶向乳腺CSCs的小分子，该方法利用最近发现的乳腺CSCs与上皮-间充质转分化之间的联系。在初步实验中，我们进行了原理验证的中试高通量筛选，成功地鉴定了一种对乳腺CSCs具有特定毒性的新化合物。筛查分析是稳健、灵敏和直接的。重要的是，二次筛查使用的细胞系试剂与初筛试验中使用的细胞系试剂基本上是同源的。这最大限度地减少了发现与感兴趣的过程无关的虚假化合物命中的可能性，从而提供了大多数屏幕中没有的非常有用的优势。试点筛查的成功使人们非常有信心地相信，如果获得资金，拟议的筛查将为基础研究和针对乳腺CSCs的潜在疗法提供有用的试剂。 公共卫生相关性：乳腺癌干细胞负责驱动肿瘤生长，目前对此知之甚少。乳腺癌干细胞对当前的抗癌治疗也具有耐药性，这表明它们是有效治疗的主要障碍。我们建议与MLPCN合作，采用一种新的、有效的基于高通量细胞的筛选来识别乳腺癌干细胞的选择性抑制物。

DESCRIPTION (provided by applicant): There is an important unmet need to identify small-molecules that can target breast cancer stem cells (CSCs). CSCs are responsible for driving tumor growth and metastasis. CSCs are also known to be resistant to most conventional therapies, including chemo- and radiation-therapy, clearly underscoring the need for therapies that target CSCs. The objective of the proposed study is to identify small-molecule probes that are selectively toxic to breast CSCs. Such molecules would be immensely useful to probe the biology of CSCs and would also serve as promising therapeutic drug candidates. We propose to screen for small molecules that target breast CSCs using a novel method we have developed that takes advantage of a recently discovered connection between breast CSCs and epithelial-mesenchymal transdifferentiation. In preliminary experiments, we have conducted a proof-of-principle pilot high-throughput screen that successfully led to the identification of a novel compound with specific toxicity for breast CSCs. The screen assay is robust, sensitive, and straightforward. Importantly, the secondary assay for the screen uses a cell-line reagent that is essentially isogenic with the cell line-reagent used from the primary screening assay. This minimizes the likelihood of find spurious compound hits unrelated to the process of interest, thereby providing a very useful advantage not present in most screens. The success of the pilot screen gives a very high degree of confidence that the proposed screen, if funded, would provide useful reagents for basic research and potential therapies targeted breast CSCs. PUBLIC HEALTH RELEVANCE: Breast cancer stem cells are responsible for driving tumor growth and are currently poorly understood. Breast cancer stem cells are also resistant to current anti-cancer treatments, indicating that they are major barriers to effective treatment. We propose to collaborate with the MLPCN to employ a novel, validated high-throughput cell-based screen to identify selective inhibitors of breast cancer stem cells.