Epithelial matrix in carcinoma development

描述(由申请人提供)： 综述：上皮基质相互作用在肿瘤侵袭和组织中的重要作用 形态发生才刚刚开始被理解。通过我们在前一个资助周期中的努力产生的数据表明，层粘连蛋白5、骨形态发生蛋白1同工酶和α6β4整合素在鳞状细胞癌(SCC)进展中起着重要作用，而层粘连蛋白10在基底细胞癌(BCC)发展中起着重要作用。目前的研究旨在进一步确定细胞外基质在非黑色素瘤皮肤癌发展中的作用。通过与其他项目的协同合作，这些研究的最终应用将是发现抑制癌症发展的新分子方法。他们建议：1.确定层粘连蛋白5和β4整合素在鳞癌进展中的作用。这一目标将确定层粘连蛋白5和β4整合素影响鳞状细胞癌肿瘤的机制。他们假设层粘连蛋白5可能抑制鳞状细胞癌的进展，而b4整合素可能促进鳞癌的进展。这一目标将利用RAS驱动的角质形成模型，在该模型中，角质形成细胞被改造成免疫缺陷小鼠的SCC样肿瘤。通过体外和体内细胞侵袭和肿瘤侵袭试验，将在体内观察层粘连蛋白5和β4整合素基因关键结构域的定点突变对鳞癌进展的影响。将评估肿瘤的大小、形态、分化、细胞凋亡和与宿主间质的粘附性。 2.阐明BMP I同工酶在鳞癌发生发展中的作用。他们的初步结果表明，BMP-1同工酶介导层粘连蛋白5的加工支持SCC肿瘤的体外侵袭，并表征了一种有效且特异的BMP-1抑制剂，具有潜在的肿瘤抑制作用。为了达到这个目的，他们试图将这些结果扩展到鳞状细胞癌肿瘤发展的体内模型。四种BMP-1同工酶的表达和层粘连蛋白5的加工将在鳞癌肿瘤的调查中进行评估。BMP-1同工酶介导层粘连蛋白5的加工对鳞癌肿瘤进展的影响将通过过表达研究来研究。最后，将在体内评估特定的BMP-1同工酶抑制剂对肿瘤侵袭的影响。 3.了解层粘连蛋白对BCC发展的影响。层粘连蛋白在组织形态发生和肿瘤发展中起着重要作用。他们的初步结果表明，层粘连蛋白10在毛发发育中起着重要作用，并表明层粘连蛋白10也支持基底细胞癌的发育。更多研究表明，层粘连蛋白5可能对基底细胞癌有抑制作用。他们将在BCC动物模型中评估层粘连蛋白10抑制作用的研究中的一些肿瘤参数。将使用多种方法，包括使用抑制性抗体和使用层粘连蛋白5小鼠。他们将通过向原代和缺失EB角质形成细胞传递层粘连蛋白5的cDNA，在动物模型中评估层粘连蛋白5的过度表达和低表达对基底细胞癌发展的影响。

DESCRIPTION (provided by applicant): Overview: The important roles that epithelial matrix interactions play in tumor invasion and tissue morphogenesis are just beginning to be understood. Data generated through our efforts during the previous funding cycle suggest important roles for laminin 5, bone morphogenic protein (BMP 1) isozymes and alpha 6 beta 4 integrin in squamous cell carcinoma (SCC) progression and an important role for laminin 10 in basal cell carcinoma (BCC) development. The current studies are directed at further defining the role of the extracellular matrix in development of non melanoma skin cancers. Through synergistic collaboration with the other Projects, the ultimate application of these studies will be to discover new molecular approaches to inhibit carcinoma development. They propose to: 1. Determine the roles of laminin 5 and beta 4 integrin in SCC progression. This Aim will determine the mechanisms by which laminin 5 and beta 4 integrin impact upon SCC tumors. They hypothesize that laminin 5 may inhibit and b4 integrin may promote SCC progression. This aim will utilize a Ras driven turnorigenesis model where keratinocytes are engineered to form SCC like tumors in immunodeficient mice. The effects of site directed mutagenesis of key domains on laminin 5 and beta 4 integrin cDNA will be examined in vivo for effects on SCC progression by in vitro and in vivo assays of cell and tumor invasion. Tumor size, morphology, differentiation, apoptosis and adhesion to host stroma will be evaluated. 2. Demonstrate the function of BMP I isozymes in SCC progression. Their preliminary results showed that BMP 1 isozyme mediated processing of laminin 5 supported SCC tumor invasion in vitro and characterized a potent and specific BMP 1 inhibitor with potential tumor inhibitory effects. In this Aim, they seek to extend these results to in vivo models of SCC tumor development. Expression of each of the four BMP 1 isozymes and the processing of laminin 5 will be evaluated in a survey of SCC tumors. The effect of BMP 1 isozyme mediated laminin 5 processing on SCC tumor progression will be studied through overexpression studies. Finally, specific BMP 1 isozyme inhibitors will be evaluated in vivo for their effects on tumor invasion. 3. Discover how laminins impact upon BCC development. Laminins perform important roles in tissue morphogenesis as well as carcinoma development. Their preliminary results show an important role for laminin 10 in hair development and suggest that laminin 10 supports BCC development as well. Additional studies suggest that laminin 5 may have an inhibitory effect on BCC. They will evaluate a number of tumor parameters in the study of the effects of laminin 10 inhibition in a BCC animal model. Multiple approaches will be utilized including the use of inhibitory antibodies and the use of laminin 5 mice. They will assess the effects of laminin 5 overexpression and underexpression on BCC development in an animal model using laminin 5 cDNAs delivered to primary and laminin 5 null EB keratinocytes.