NOVEL SERPIN INHIBITOR OF ORAL SQUAMOUS CARCINOMA

描述(由申请人提供)：本提案试图测试的假设是，CBP2/Hsp47构成调控复杂性的一部分，该复杂性决定了XVI11胶原(COL18)是被生产和加工成作为促动素还是作为内皮抑素(ES)发挥功能。此外，CBP2/Hsp47和COL18形成了血管抑制-血管生成转换轴的组成部分，确保了肿瘤的生长和促进侵袭。这一建议试图证明在常氧条件下，COL18在CBP2/HSP47的帮助下在可能的地方产生和分泌，或被基质金属蛋白酶加工，以促进肿瘤细胞的运动和侵袭。然而，在缺氧或坏死导致的低微环境pH条件下，COL18被组织蛋白酶L降解，除非被胞外Cbp2/Hsp47抑制为ES。内皮抑素抑制肿瘤细胞的运动和侵袭，直到促血管生成信号建立重建正常氧的新生血管。本研究将通过完成以下具体目标来完成：1)利用体外模型证明短型胶原XVIII(缺少卷曲结构域)促进肿瘤细胞的运动和侵袭。结果表明，XVilI胶原蛋白水解区，特别是三聚化的NC1区和基质金属蛋白酶活性后产生的片段同样具有促动生作用，而其羧基末端内皮抑素结构域则同时抑制运动和侵袭；2)确定CBP2/Hsp47是否构成调控复杂性的一个重要特征，通过在口腔鳞癌细胞系中过表达或敲除CBP2/Hsp47来控制XVil型胶原；3)确定XVil型胶原的信号通路。促进肿瘤细胞侵袭的反馈机制(S)，通过它的C末端片段，内皮抑素，作为拮抗剂作用；4)在实践中证明，在实体人类肿瘤中，CBP2/HSP47和微细胞环境构成了调控复杂性的重要特征，通过调节XVIII胶原的产生和处理来控制肿瘤细胞的生长和侵袭模式。这些研究将有助于将正常和异常细胞的基因转录谱、表达蛋白质及其活性与功能状态相关联。

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.