NOVEL SERPIN INHIBITOR OF ORAL SQUAMOUS CARCINOMA

描述（由申请人提供）：本提案试图检验的假设是，CBP 2/Hsp 47构成了决定胶原XVIII（col 18）是否产生和加工以发挥运动原或内皮抑制素（ES）功能的调节复杂性的一部分。此外，CBP 2/HSP 47和col 18形成了血管生成抑制-血管生成转换轴的组成部分，其确保肿瘤生长并促进侵袭。该建议试图证明在常氧期间，col 18在CBP 2/HSP 47的帮助下产生和分泌，其中col 18本身或被基质金属蛋白酶加工成运动片段促进肿瘤细胞运动和侵袭。然而，在由缺氧或坏死引起的低微环境pH下，col 18被组织蛋白酶L加工成ES，除非被细胞外CBP 2/Hsp 47抑制。内皮抑制素抑制肿瘤细胞运动和侵袭，直到促血管生成信号建立新血管系统，重新建立正常氧。所提出的研究将通过完成以下具体目标来完成：1）使用体外模型证明短形式的胶原XVIII（缺乏卷曲结构域）促进肿瘤细胞运动和侵袭。然后表明胶原蛋白XVilI的蛋白水解片段，特别是三聚NC 1结构域和基质金属蛋白酶活性后产生的片段同样具有运动原性，而羧基末端、内皮抑制素结构域抑制运动性和侵袭性; 2）确定CBP 2/Hsp 47是否构成通过过表达或敲除CBP 2/Hsp 47来控制胶原蛋白XVII 11的调节复杂性的重要特征。Hsp 47在人口腔鳞癌肿瘤细胞系中的表达; 3）确定胶原XVII！促进肿瘤细胞侵袭和反馈机制，通过该反馈机制，其C-末端片段内皮抑制素起拮抗剂的作用，和4）在实践中提供证据，即在实体人肿瘤中，CBP 2/HSP 47和微细胞环境构成了通过调节胶原XVIII的产生和加工来控制肿瘤细胞生长和侵袭模式的调节复杂性的重要特征。这些研究将有助于将基因转录谱、表达的蛋白质库及其活性与正常和异常细胞的功能状态相关联。

DESCRIPTION (provided by applicant): The hypothesis that this proposal seeks to test is that CBP2/Hsp47 constitutes a portion of the regulatory complexity that determines whether collagen XVIll (col 18) is produced and processed to function as a motogen or as an endostatin (ES). Moreover, that CBP2/Hsp47 and col 18 form an integral component of the angiostatic-angiogenic switching axis that ensures tumor growth and promotes invasion. This proposal seeks to prove that during normoxia col 18 with the aid of CBP2/Hsp47 is produced and secreted where it may itself or processed by matrix metalloproteinases to motogenic fragments promote tumor cell motility and invasion. However, at low microenvironmental pH resulting from hypoxia or necrosis, col 18 is processed by cathepsin L, unless inhibited by extracellular CBP2/Hsp47, to ES. Endostatin inhibits tumor cell motility and invasion until and proangiogenic signaling establishes a neovasculature that reestablishes normoxia. The proposed research will be accomplished through the completion of the following specific aims: 1) demonstrate that short form of collagen XVIII (lacking the frizzled domain) promote tumor cell motility and invasion using in vitro models. Then show that proteolytic fragments of collagen XVilI, particularly the trimerized NC1 domain and fragments produced following matrix metalloproteinase activity are likewise motogenic while the carboxyl terminus, endostatin domain, inhibits both motility and invasion; 2) determine whether CBP2/Hsp47 constitutes an important feature of the regulatory complexity that controls collagen XVill by over expressing or knocking out CBP2/Hsp47 in human oral squamous carcinoma tumor cell lines; 3) Determine the signaling pathways by which collagen XVII! promotes tumor cell invasion and the feedback mechanism(s) by which its' C-terminal fragment, endostatin, functions as an antagonist and 4 )provide the proof in practice that in solid human tumors CBP2/Hsp47 and the microcellular environment constitute important features of the regulatory complexity that control patterns of tumor cell growth and invasion by regulating the production and processing of collagen XVIII. These studies will aid in correlating the gene transcription profiles, and repertoire of proteins expressed and their activities with functional status of both normal and aberrant cells.