SMRT-2 in Epidermal Neoplasia

描述（由申请方提供）：上皮恶性肿瘤代表了绝大多数人类癌症。了解这些癌症的发展和进展对于确定新的治疗靶点至关重要。鳞状细胞癌（SCC）是第二常见的皮肤癌，其结果通常与分化状态呈负相关。为了鉴定表皮分化和肿瘤形成的长链非编码RNA（lncRNA）调节因子，我们进行了RNA测序并鉴定了新的推定lncRNA SMRT-2。与正常患者和部位匹配对照相比，SMRT-2在所有12个测试的肿瘤样品中下调。SMRT-2也在表皮分化过程中上调，初步数据表明它是这一过程所必需的。 首先，我们将产生和表征SMRT-2表达改变的器官型人表皮肿瘤。我们将使用的模型与人类SCC非常相似，并且适合遗传操作[9]。我们已经设计了实验，以消耗SMRT-2和强制SMRT-2表达在这个模型的上下文中。这些实验将使我们能够评估SMRT-2促进肿瘤发生和减少肿瘤分化的能力，以及评估其作为肿瘤抑制因子在抑制肿瘤侵袭和细胞增殖中的潜在作用。总的来说，这些研究将使我们能够确定SMRT-2在表皮瘤形成中的具体作用。 LncRNA的功能机制以前已经通过寻找相互作用伴侣来阐明[14]。为了进一步表征SMRT-2的机制，我们接下来将通过鉴定其蛋白质、RNA和DNA相互作用伴侣来表征SMRT-2相互作用组。蛋白质相互作用伴侣将通过使用蛋白质微阵列芯片来鉴定，标记的SMRT-2将与该芯片杂交[32]。RNA和DNA相互作用伴侣将通过SMRT-2的RNA下拉来鉴定，随后将对相互作用的转录物[16]或基因组DNA [33]进行测序。这些研究将在表皮分化和器官型表皮瘤形成的背景下进行，以识别可能在癌症中紊乱的SMRT-2相互作用。此外，了解SMRT-2相互作用组将使我们能够了解SMRT-2在细胞生物分子背景下的作用机制。有了这些知识，SMRT-2或其相互作用伙伴之一可以用作表皮瘤形成的生物标志物或治疗靶点。 在拟议的资助期结束时，我们希望能够描述SMRT-2在表皮肿瘤进展中的作用，并通过其相互作用伙伴确定潜在的作用机制。这些结果可能阐明用于治疗SCC和其他上皮癌的其他治疗靶点。

DESCRIPTION (provided by applicant): Epithelial malignancies represent the vast majority of human cancers. Understanding the development and progression of these cancers is essential in the effort to identify new therapeutic targets. Squamous cell carcinoma (SCC) is the second most common skin cancer, with outcomes that are often inversely correlated with differentiation state. To identify long non-coding RNA (lncRNA) regulators of both epidermal differentiation and neoplasia, we performed RNA-Sequencing and identified the novel, putative lncRNA SMRT-2. SMRT-2 is down-regulated in all 12 tumor samples tested compared to normal patient and site matched controls. SMRT-2 is also up-regulated over the course of epidermal differentiation and preliminary data indicates it is required for this process. First, we will generate and characterie organotypic human epidermal neoplasia with altered SMRT-2 expression. The model we will use closely resembles human SCC and is amenable to genetic manipulation [9]. We have designed experiments to both deplete SMRT-2 and force SMRT-2 expression in the context of this model. These experiments will allow us to assess the ability of SMRT-2 to promote tumorigenesis and diminish neoplastic differentiation, as well as evaluate its potential role as a tumor suppressor in inhibiting neoplastic invasion and cell proliferation. Overall, these studies will allow us to identify the specific role of SMRT-2 in epidermal neoplasia. LncRNA functional mechanisms have previously been elucidated by looking to interaction partners [14]. To further characterize the mechanism of SMRT-2, we will next characterize the SMRT-2 interactome by identifying its protein, RNA, and DNA interaction partners. Protein interaction partners will be identified by using a protein microarray chip to which labeled SMRT-2 will be hybridized [32]. RNA and DNA interaction partners will be identified by RNA pulldown of SMRT-2 and interacting transcripts [16] or genomic DNA [33] will be subsequently sequenced. These studies will be conducted in the context of both epidermal differentiation and organotypic epidermal neoplasia to allow identification of SMRT-2 interactions that might be deranged in cancer. Additionally, knowing the SMRT-2 interactome will allow us to understand the mechanism of SMRT-2 action in the context of cellular biomolecules. With this knowledge, either SMRT-2 or one of its interaction partners could be used as a biomarker or therapeutic target of epidermal neoplasia. At the end of the proposed funding period, we hope to have characterized the role of SMRT-2 in epidermal tumor progression and identified a potential mechanism of action via its interaction partners. These results may illuminate additional therapeutic targets for the treatment of SCC and other epithelial cancers.