How KAI1/CD82 Suppresses Cancer Invasion and Metastasis

描述(申请人提供)：KAI1/CD82是实体瘤的转移抑制因子。KAI1/CD82介导的转移抑制机制在很大程度上仍不清楚。最近的研究表明，KAI1/CD82可以诱导脂筏和富含Tetraspanin的微区等膜微域的重组，减弱生长因子和整合素信号，抑制细胞突起和回缩。值得注意的是，我们还发现KAI1/CD82通过抑制癌细胞侵袭来抑制肿瘤转移。因此，KAI1/CD82如何抑制癌细胞运动成为了解KAI1/CD82如何抑制肿瘤转移的突出问题。我们推测KAI1/CD82通过重组膜微区抑制癌细胞的迁移和侵袭，从而减少突起和收缩过程以及由外而内的有丝分裂信号。为了阐明KAI1如何抑制肿瘤的侵袭和转移，我们将首先对含有KAI1/CD82的透射电子显微镜进行结构和功能的研究，包括：1)鉴定CD82分子中与脂筏物理连接的结构元件(S)；2)系统地表征CD82阳性的透射电子显微镜的蛋白质和脂质成分；其次，我们将通过研究含有CD82的透射电子显微镜如何使细胞膜弯曲和运输来确定含有KAI1/CD82的透射电子显微镜调节细胞运动活动的机制。我们还将评估CD82诱导的膜曲率改变和转运在i)膜运动活性、ii)有丝分裂信号和iii)CD82介导的抑制癌症侵袭中的作用。最后，我们将从以下几个方面确定KAI1/CD82抑制体内肿瘤侵袭的机制：i)CD82抑制体内肿瘤侵袭的细胞运动活性；ii)与CD82物理或功能相关的蛋白质对CD82介导的肿瘤侵袭的抑制作用；以及iii)上述分析的CD82功能是否对其抑制体内癌症侵袭至关重要。综上所述，这项研究将使我们能够了解KAI1/CD82作为膜微区的组织者如何调控细胞迁移、肿瘤侵袭和肿瘤转移，以及ii)揭示细胞运动和肿瘤转移调控的新机制，即膜弯曲和运输调节侵袭和转移。通过对KAI1/CD82在体外和体内机制的深入研究，我们将对肿瘤的侵袭和转移有一个完整的认识，最终将使KAI1/CD82成为肿瘤侵袭和转移的诊断标志和治疗靶点。

DESCRIPTION (provided by applicant): KAI1/CD82 is a metastasis suppressor of solid tumors. The mechanism of KAI1/CD82- mediated metastasis suppression still remains largely elusive. Recent studies indicate that KAI1/CD82 induces the reorganization of membrane microdomains such as lipid rafts and tetraspanin-enriched microdomain (TEM), attenuates growth factor and integrin signaling, and inhibits cell protrusion and retraction. Notably, we also found that KAI1/CD82 suppresses cancer metastasis by inhibiting cancer cell invasiveness. Thus, how KAI1/CD82 inhibits cancer cell movement becomes the outstanding question to understand how KAI1/CD82 suppresses cancer metastasis. We hypothesize that KAI1/CD82 inhibits cancer cell migration and invasion through re-organizing membrane microdomains and consequently reducing protrusive and retraction processes and the outside-in mitogenic signaling. To elucidate how KAI1 suppresses cancer invasion and metastasis, we will first carry out the structural and functional characterization of KAI1/CD82-containing TEM by i) identifying the structural element(s) in CD82 molecule that physically links TEM to lipid rafts, ii) systematically characterizing the protein and lipid components of CD82-positive TEM, and iii) analyze the trafficking and subcellular localization of CD82- containing TEM. Secondly, we will determine the mechanism by which KAI1/CD82-containing TEM regulates the cellular motile activities by addressing how CD82-containing TEM renders membrane curvature and trafficking. We will also assess the roles of CD82-induced alterations in membrane curvature and trafficking in i) membrane motile activities, ii) mitogenic signaling, and iii) CD82-mediated suppression of cancer invasion. Finally, we will determine the mechanism by which KAI1/CD82 inhibits cancer invasion in vivo by addressing i) through which cellular motile activity CD82 suppresses cancer invasion in vivo, ii) the contributions of the proteins physically or functionally associated with CD82 to CD82-mediated suppression of cancer invasion; and iii) whether the CD82 features analyzed above are crucial for its suppression of cancer invasion in vivo. Together, the proposed study will enable us to i) understand how KAI1/CD82 regulates cell migration, cancer invasion, and cancer metastasis as an organizer for membrane microdomains and ii) unveil a novel mechanisms by which cell motility and cancer metastasis are regulated, i.e., membrane curvature and trafficking regulates invasion and metastasis. From the in-depth in vitro and in vivo mechanistic study of KAI1/CD82, we will develop an integrated understanding of cancer invasion and metastasis, which will ultimately lead to the development of KAI1/CD82 into a diagnostic marker and therapeutic target for cancer invasion and metastasis.