Roles of long non-coding RNAs in human normal and neoplastic epidermis

该提案旨在深入了解人类正常和肿瘤皮肤中长链非编码RNA（lncRNA）的功能相关性。人类基因组编码数千个长度>200个碱基对的非蛋白质编码转录物。尽管最近的研究表明lncRNA在多种生物学过程中发挥重要作用，但它们对控制癌症发展期间成熟人体组织转变为肿瘤状态的影响在很大程度上仍然未知。通过本文提出的项目，我计划使用人类器官型表皮作为模型系统来表征6种皮肤癌相关lncRNA对肿瘤进展的影响。基于先前的深度RNA测序分析，选择了所有6种lncRNA用于本研究，并且在人皮肤癌标本（鳞状细胞癌; SCC）中被抑制以及在分化期间富集。因此，这些lnRNA在控制表皮组织分化以及SCC发展中具有双重作用的高度倾向，并且将通过2个广泛的目的进行功能分析：目的I旨在表征SCC相关lncRNA对表皮分化的功能影响。我们将使用RNA干扰产生所有6种SCC相关lncRNA缺陷的人表皮。然后将这些组织用于检查这些lncRNA对表皮分化的功能作用。在目的II中，我们将描述功能性SCC相关lncRNA对人类组织肿瘤的影响。为此，我们将产生持续表达SCC相关lncRNA的器官型肿瘤组织，这些lncRNA被证明对人类表皮分化具有重要功能（如目标I中所测试）。然后，我们将使用这些肿瘤组织来分析持续lncRNA表达对肿瘤细胞生长和侵袭率的影响。对癌症中被错误调控的长链非编码RNA的表征将进一步深入了解lncRNA在成熟人体组织分化和肿瘤形成中的功能和机制，并可能为预防和治疗表皮稳态障碍以及皮肤癌提供新的靶点。

This proposal seeks insight into the functional relevance of long non-coding RNAs (lncRNAs) in human normal and neoplastic skin. The human genome encodes several thousand long non-protein coding transcripts >200 base pairs in length. Although recent studies have shown that lncRNAs play important roles in a variety of biological processes, their impact on controlling the transition of mature human tissue into a neoplastic state during cancer development remains largely unknown. With the projects proposed here, I plan to characterize the impacts of 6 skin cancer-associated lncRNAs on neoplastic progression using human organotypic epidermis as a model system. All 6 lncRNAs were selected for this study based on previous deep RNA sequencing analyses, and were repressed in human skin cancer specimens (squamous cell carcinoma; SCC) as well as enriched during differentiation. Thus, these lnRNAs have high propensity for a dual-role in control of epidermal tissue differentiation as well as SCC- development and will be functionally analyzed through 2 broad aims:Aim I is designed to characterize the functional impact of SCC-associated lncRNAs on epidermal differentiation. We will generate human epidermis deficient for all of the 6 SCC-associated lncRNAs using RNA interference. These tissues will then be used to examine functional roles of these lncRNAs on epidermal differentiation. In Aim II, we will characterize the impacts of functional SCC-associated lncRNAs on human tissue neoplasia. For this, we will generate organotypic, neoplastic tissue with sustained expression of SCC-associated lncRNAs that prove to be functionally important for human epidermal differentiation (as tested in Aim I). We will then use these neoplastic tissues to analyze the impacts of sustained lncRNA expression on tumor cell growth and invasion rate. Characterization of long non-coding RNAs that are mis-regulated in cancer will gain further insight into functions and mechanisms of lncRNAs in differentiation as well as neoplasia of mature human tissue, and may provide novel targets for prevention and treatment of disorders of epidermal homeostasis as well as skin cancer.