METASTATIC REGULATION OF DIFFERENTIAL SPLICING OF CD44

这是一个香农奖提供部分支持的研究 项目不属于指定机构的资助范围，但 在优秀的边缘。香农奖旨在提供 支持测试方法的可行性;支持进一步测试 和完善研究技术;进行二次分析或提供 数据集;或进行可以证明PI的离散项目 研究能力或为已经优秀的 应用程序.以下摘要摘自原始文件 由主要经销商提交。 描述（改编自研究者摘要）：研究期间 转移时，肿瘤细胞脱离原发肿瘤块并通过 通过细胞外基质侵入身体的其他部位。 的 肿瘤细胞穿透基质并相互附着的能力 部分是由于肿瘤细胞上存在细胞表面蛋白 与细胞外基质或细胞表面相互作用 其他细胞的蛋白质。 一种这样的细胞表面蛋白， 细胞外结构域与转移潜能相关， 多功能粘附糖蛋白，CD 44，表达于 哺乳动物中的各种细胞。 来自CD 44基因的RNA经历 广泛的选择性剪接。 该基因含有10个内部盒 可选择地包括以产生CD 44的变体形式的外显子 有额外的细胞外结构域， 携带CD 44的细胞的特异性。 包括一个特定的子集 选择性外显子与转移扩散有很强的相关性 在结肠癌、乳腺癌、卵巢癌和脑癌中 CD 44的变异形式现在被用来评估肿瘤的转移潜能， 人类癌症 含有交替包含的外显子的CD 44 cDNA的表达 与转移相关的基因足以使同基因的 肿瘤细胞和针对CD 44变体形式的抗体可防止转移 在动物模型中传播。 因此，剪接模式的改变 CD 44 mRNA的表达足以使肿瘤细胞转移。 这 该提案旨在了解参与的分子机制， 卵巢癌和乳腺癌转移中CD 44选择性剪接 癌肿瘤细胞系和比较转移性替代 剪接到白细胞生长刺激期间发生的剪接，或 上皮细胞和早期发育中的组织形成期间。 具体目标包括：1）确定顺式作用序列， 调节CD 44替代外显子的包含。 这些目标 实验是区分由于组织的剪接调节- 由于基因组的改变， 一般拼接机械的水平或活动; 2）确定 的交易所需的因素，以规管列入的 转移性细胞与正常细胞中的CD 44替代外显子; 3） 选择性剪接事件与细胞凋亡关系的研究 产生CD 44转移性变体和正常组织建立 在哺乳动物的早期发育中。

THIS IS A SHANNON AWARD PROVIDING PARTIAL SUPPORT FOR THE RESEARCH PROJECTS THAT FALL SHORT OF THE ASSIGNED INSTITUTE'S FUNDING RANGE BUT ARE IN THE MARGIN OF EXCELLENCE. THE SHANNON AWARD IS INTENDED TO PROVIDE SUPPORT TO TEST THE FEASIBILITY OF THE APPROACH; DEVELOP FURTHER TESTS AND REFINE RESEARCH TECHNIQUES; PERFORM SECONDARY ANALYSIS OR AVAILABLE DATA SETS; OR CONDUCT DISCRETE PROJECTS THAT CAN DEMONSTRATE THE PI'S RESEARCH CAPABILITIES OR LEND ADDITIONAL WEIGHT TO AN ALREADY MERITORIOUS APPLICATION. THE ABSTRACT BELOW IS TAKEN FROM THE ORIGINAL DOCUMENT SUBMITTED BY THE PRINCIPAL INVESTIGATOR. DESCRIPTION (adapted from the investigator's abstract): During metastasis, tumor cells break away from the primary tumor mass and pass through the extracellular matrix to invade other areas of the body. The ability of tumor cells to penetrate the matrix and attach to each other is, in part, due to the presence of cell surface proteins on tumor cells that interact with either the extracellular matrix or with cell surface proteins on other cells. One such cell surface protein whose extracellular domains have been correlated with metastatic potential is the multifunctional adhesion glycoprotein, CD44, which is expressed in a wide variety of cells in mammals. RNA from the CD44 gene undergoes extensive alternative splicing. The gene contains 10 internal cassette exons that are alternatively included to produce variant forms of CD44 with additional extracellular domains that influence the binding specificity of cells bearing CD44. Inclusion of a specific sub-set of the alternative exons has a strong correlation with metastatic spread in colon, breast, ovarian and brain cancer such that expression of variant forms of CD44 is now used to assess metastatic potential of human cancers. Expression of CD44 cDNAs containing the alternatively-included exons associated with metastasis is sufficient to confer metastasis on syngenic tumor cells and antibodies to the variant form of CD44 prevent metastatic spread in animal models. Therefore, alteration in the splicing pattern of CD44 mRNA is sufficient to render a tumor cell metastatic. This proposal is aimed at understanding the molecular mechanism involved in the alternative splicing of CD44 during metastasis of ovarian and breast cancer tumor cell lines and comparing the metastatic alternative splicing to that occurring during growth stimulation of leukocyte or epithelial cells and during tissue formation in early development. Specific aims include: 1) determination of the cis-acting sequences that regulate inclusion of the CD44 alternative exons. The goal of these experiments is to distinguish regulation of splicing due to tissue- specific factors from regulation of splicing due to alterations in the levels or activities of the general splicing machinery; 2) determination of the trans-acting factors required for regulation of inclusion of the CD44 alternative exons in metastatic versus normal cells; 3) Investigation of the relationship between the alternative splicing event producing the CD44 metastatic variants and normal tissue establishment during early mammalian development.