COBRE PROJ 8: ROLES OF IKK ?LPHA IN SKIN DEVELOPMENT AND DYSPLASIA

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 李秋堂，PI 该项目的长期目标是阐明IKK α对表皮稳态发挥作用的细胞和分子机制。IKK α是NF-κ B信号通路中IKK复合物的亚基。IKK α敲除小鼠在出生时死于与角质形成细胞增殖增加和终末分化受损相关的皮肤缺陷。 IKK α在角质形成细胞中的功能不依赖于其激酶活性和NF-κ B信号通路。 IKK α调节角质形成细胞增殖和分化的机制尚不清楚。 在本研究中，我们将测试的假设，IKK α调节角质形成细胞的细胞周期和功能作为一个肿瘤抑制基因在皮肤中。 具体而言，将解决以下问题：（1）IKK α在角质形成细胞细胞周期和细胞周期检查点的调节中的作用是什么？ (2)角质形成细胞中与IKKa相互作用的蛋白质有哪些？ (3)这些相互作用的蛋白质是如何参与角质形成细胞中IKK α信号转导的？ (4)IKK α在皮肤中作为肿瘤抑制因子发挥作用吗？ 将使用遗传学、细胞学和生物化学方法的组合，在胚胎第18.5天从野生型或IKK α缺陷胚胎中分离的原代角质形成细胞中进行细胞周期分析和功能研究。 将进一步验证和表征IKK α免疫沉淀和质谱法产生的数据，以在分子水平上了解IKK α的功能。 我们将检查暴露于致突变致癌物的IKK α +/-小鼠的皮肤肿瘤易感性和转移潜力，并检查IKK α +/-小鼠肿瘤中IKK α基因座的杂合性丢失频率。 这些知识对于阐明皮肤癌等临床相关问题非常重要。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Qiutang Li, PI The long-term goal of this project is to elucidate the cellular and molecular mechanism by which IKKalpha exerts its effects on epidermal homeostasis. IKKalpha is a subunit of IKK complexes in the NF-kappa B signaling pathway. IKKalpha knockout mice die at birth from the skin defects associated with increased proliferation and impaired terminal differentiation in keratinocytes. The function of IKKalpha in keratinocytes is independent from its kinase activity and NF-kappa B signaling pathway. The mechanism by which the IKKalpha regulates keratinocyte proliferation and differentiation remains unclear. In the present studies, we will test the hypothesis that IKKalpha regulates keratinocyte cell cycle and functions as a tumor suppressor gene in skin. Specifically, the following questions will be addressed: (1) What are the roles of the IKKalpha in regulation of the keratinocyte cell cycle and cell cycle checkpoint? (2) What are the IKKalpha-interacting proteins in keratinocyte? (3) How do those interacting proteins participate in the IKKalpha signaling in the keratinocytes? (4) Does IKKalpha function as a tumor suppressor in skin? Cell cycle analysis and functional studies will be performed in the primary keratinocytes isolated from either wild-type or the IKKalpha deficient embryos at embryonic day 18.5 using a combination of genetics, cytological, and biochemical approaches. The data that have been generated from IKKalpha immunoprecipitation and mass spectrometry will be further validated and characterized to understand the IKKalpha function at a molecular level. We will examine skin tumor susceptibility and metastatic potential in IKKalpha+/- mice exposed to mutagenic carcinogen and check the loss of heterozygosity frequency at the IKKalpha locus in IKKalpha+/- mouse tumors. The knowledge will be important for elucidating clinically relevant problems such as skin cancers.