Neuropilin and Semaphorin Function in Development and Tumor Angiogenesis

我们的实验室长期以来一直对识别分子和理解 调节血管生成、肿瘤生长和进展的机制。我们最近报道了 在患有乳腺癌的女性的尿液中存在ADAMT 2（解整合素和金属蛋白酶12）， 进一步证明，它的存在可以预测这些女性的临床状态和分期。在这 同样的研究，我们首次报道，ADAM 12可以降解细胞外基质成分， 包括IV型胶原蛋白和纤连蛋白，并且它也具有明胶酶活性。这些数据让我们 假设ADAM 12可能通过重塑肿瘤血管生成和进展， 细胞外基质和/或通过各种其他机制，已归因于这个家庭的 酶，包括胞外域脱落，调节生长因子的可用性和介导细胞-细胞， 细胞-基质相互作用我们最近还证明，ADAM 12水平显著高于对照组。 在肿瘤进展过程中最早的检查点之一，血管生成开关， 其在人乳腺癌细胞中的过表达导致VEGF产生的显著增加。 ADAM 12过表达还导致人乳腺癌细胞运动性的显著增加， 侵袭性，经典间充质标志物波形蛋白和纤连蛋白的上调， 散射表型，所有这些都与EMI的诱导一致（上皮细胞到间质细胞）。 转化），这是癌症进展的标志。ADAM 12也赋予雌激素非依赖性（ER） ER阳性乳腺癌细胞的生长状态。最后，在一系列的初步研究中，我们发现， ADAM 12过表达促进原位人乳腺癌的体内生长。范围内 我们的提案的具体目标，我们将确定ADAM 12可能调节的机制 血管生成、肿瘤生长和转移。这些机械研究将得到一系列 在体内实验中，我们将确定ADAM 12对人类乳腺肿瘤生长的影响， 使用三种互补的体内模型进行进展。这些研究也将提供信息， ADAM 12作为治疗靶点在乳腺癌及其进展中的潜在作用。在 本研究的第三个目的是，我们将确定尿中的ADAM 12，单独或与其他癌症联合， 生物标志物可以是乳腺癌及其进展的诊断和/或预后生物标志物。这些 建议在以下具体目标的范围内进行研究： 1.为了确定ADAM 12调节血管生成、肿瘤生长和肿瘤生长的机制， 转移 2.确定ADAM 12是否促进人乳腺癌的体内生长和进展 3.为了确定是否存在ADAM 12，单独或与其他泌尿系癌症的多重 生物标志物，预测肿瘤的存在，进展和治疗效果的动物模型， 乳腺癌和患有乳腺癌的人类患者。 通过系统地鉴定可能在肿瘤的发展和进展中发挥作用的新分子， 乳腺癌，并通过解剖这些分子可能发挥其作用的机制， 我们将有机会开发新的和改进的治疗、诊断和预后策略 可以显著提高癌症患者的存活率。

Our laboratory has had a long standing interest in identifying the molecules and understanding the mechanisms that regulate angiogenesis, tumor growth and progression. We have recently reported the presence of ADAMT2 (A Disintegrin and Metalloproteinase 12),in the urine of women with breast cancer and have further demonstrated that its presence predicts both clinical status and stage in these women. In this same study, we reported, for the first time, that ADAM12 can degrade extracellular matrix components including type IV collagen and fibronectin and that it has gelatinase activity as well. These data have led us to hypothesize that ADAM12 may play a role in tumor angiogenesis and progression by remodeling the extracellular matrix and/or through a variety of other mechanisms that have been attributed to this family of enzymes, including ectodomain shedding, regulation of growth factor availability and mediating cell-cell and cell-matrix interactions. We have also recently demonstrated that ADAM12 levels are significantly upregulated during one of the earliest checkpoints during tumor progression, the angiogenic switch and that its overexpression in human breast cancer cells results in a significant increase in the production of VEGF. ADAM12 overexpression also resulted in a significant increase in human breast cancer cell motility and invasivity, an upregulation of the classic mesenchymal markers vimentin and fibronectin and the acquisition of a scattering phenotype, all of which are consistent with the induction of EMI (Epithelial to Mesenchymal Transformation), a hallmark of cancer progression. ADAM12 also conferred estrogen-independent(ER) growth status to ER positive breast cancer cells. Finally, in a series of preliminary studies, we found that ADAM12 overexpression promoted prthotopic human breast cancer growth in vivo. Within the context of the Specific Aims of our proposal, we will determine the mechanism(s) by which ADAM12 may be regulating angiogenesis, tumor growth and metastasis. These mechanistic studies will be complimented by a series of in vivo ones in which we will determine the effects of ADAM12 on human breast tumor growth and progression using three complimentary in vivo models. These studies will also be informative as to the potential role of ADAM12 as a therapeutic target in the treatment of breast cancer and its progression. In the third Aim of this study, we will determine whether urinary ADAM12, alone or in combination with other cancer biomarkers, may be a diagnostic and/or prognostic biomarker for breast cancer and its progression. These studies are proposed within the context of the following Specific Aims: 1. To determine the mechanism(s) by which ADAM12 may regulate angiogenesis, tumor growth and metastasis 2. To determine whether ADAM12 promotes human breast cancer growth and progression in vivo 3. To determine whether the presence of ADAM12, alone or multiplexed with other urinary cancer biomarkers, predicts tumor presence, progression, and therapeutic efficacy in animal models of breast cancer and in human patients with oreast cancer. By systematically identifying novel molecules that may be playing a role in the development and progression of breast cancer, and by dissecting the mechanisms by which such molecules may be exerting their effects, we will have the opportunity to develop new and improved therapeutic, diagnostic and prognostic strategies that could result in significantly improved cancer patient survival.