Novel Genetic Models to Study the Role of DNp63 in Squamous Cell Carcinoma

描述（由申请方提供）：鳞状细胞癌（SCC）是最常见的皮肤恶性肿瘤之一，全球发病率不断增加，因此构成了严重的公共卫生问题。尽管迄今为止，已经深入研究了导致SCC发展和进展的潜在遗传和表观遗传事件，但尚未成功开发出针对这种潜在致命形式的皮肤癌的预防或治疗策略。因此，迫切需要进行更多的研究，以更好地了解这种皮肤病的分子病因，并利用这些知识开发新的有效治疗方法。来自我们实验室和其他实验室的证据，以及来自癌症基因组图谱计划的大规模测序数据，强烈表明转录因子p63，特别是最常见的Np 63亚型，在SCC发展的早期阶段作为癌基因发挥作用。我们发现，Np 63在转基因小鼠（Np 63 BG）皮肤中的过度表达导致了一种严重的表型，该表型与SCC的早期阶段相关的许多潜在的组织学、病理学和分子特征相同。然而，我们目前关于Np 63在SCC早期阶段发挥作用的分子基础的知识仍然很少。因此，我们将利用Np 63 BG和Np 63-GFP敲入小鼠，这两种小鼠都是在我们的实验室中开发的，以解决两个重要的感兴趣的领域.首先，鉴于SCC中Np 63水平升高，我们建议通过对Np 63 BG和Np 63杂合子小鼠进行两阶段皮肤化学致癌研究，实验评估这种亚型在肿瘤发展早期的贡献。从这种体内研究中获得的数据将建立Np 63的致癌功能，并提供了一个更好的理解的具体作用的Np 63亚型在早期阶段的SCC的开始和发展，这是迄今为止还没有可能由于缺乏Np 63亚型特异性小鼠模型。其次，我们建议在全基因组范围内确定Np 63特异性靶基因和促进SCC发展的调控网络。为此，我们将进行染色质免疫沉淀（ChIP），然后进行深度测序（ChIP-seq），以确定Np 63的新的直接靶基因，这些基因在Np 63 BG小鼠皮肤中发生了改变，并且可能有助于SCC的发展。总的来说，从这些遗传学研究中收集到的信息不仅有可能显着提高我们对SCC潜在致病性改变的分子理解，而且还可能发现新的驱动途径和关键参与者，以供未来开发作为治疗选择。

DESCRIPTION (provided by applicant): Squamous cell carcinoma (SCC) is one of the most common forms of skin malignancies with increasing global incidence, thus constituting a serious public health concern. Although to date the underlying genetic and epigenetic events contributing to the development and progression of SCC have been intensively studied, no resulting preventative or therapeutic strategies have been successfully developed to target this potentially lethal form of skin cancer. Therefore there is a vital need for additional studies to better understand the molecular etiology of this skin disease and harness that knowledge for the development of new and effective treatments. Evidence from our laboratory as well as others, together with the large-scale sequencing data emerging from the Cancer Genome Atlas Project, strongly suggest that the transcription factor p63, specifically the most prevalent Np63 isoforms, function as an oncogene in the early stages of SCC development. We show that overexpression of Np63 in transgenic mouse (Np63BG) skin results in a severe phenotype that share many of the underlying histological, pathological and molecular features associated with the early stages of SCC. However, our current knowledge regarding the molecular underpinnings through which Np63 functions in the early stages of SCC remains sparse. Hence, we will utilize Np63BG and Np63-GFP knock- in mice, both of which were developed in our laboratory, to address two important areas of interest. First, given the elevated levels of Np63 in SCC, we propose to experimentally evaluate the contribution of this isoform in the early stages of tumor development by performing two-stage skin chemical carcinogenesis studies on the Np63BG and Np63 heterozygous mice. Data obtained from such in vivo studies will establish the oncogenic function of Np63 and provide a better understanding of the specific role of the Np63 isoforms in the early stages of SCC initiation and progression, which to date have not been possible due to the lack of Np63 isoform specific mouse models. Secondly we propose to identify, on a genome-wide scale, Np63 specific target genes and regulatory networks contributing to the development of SCC. Towards this end, we will perform chromatin-immunoprecipitation (ChIP) followed by deep sequencing (ChIP-seq) to identify novel direct target genes of Np63, which are altered in the Np63BG mouse skin, and which likely contribute to the development of SCC. Collectively the information gleaned from these genetic studies has the potential to not only significantly enhance our molecular understanding of the underlying pathogenic alterations of SCC but also to unearth new driver pathways and critical players for future exploitation as therapeutic options.