BIOLOGY AND PATHOLOGY OF A MODULATOR OF FGF

描述（申请人摘要）：成纤维细胞生长因子（FGF-1或FGF-2）是 在成人的大多数正常组织中以显著浓度存在。 然而，这些FGF以非活性状态固定在细胞外基质上。 基质，并且仅很少了解它们是如何溶解和活化的 到达它们的受体。这些生长因子可以通过以前的 鉴定了FGF（BP 1）的分泌结合蛋白。 我们发现，类维生素A调节的BP 1可以增强局部的 储存的、固定的FGF和BP 1的表达支持肿瘤生长， FGF-2阳性非致瘤细胞的血管生成。新近发现的一 新的BP 2显示出相似的生物活性。在初步研究中我们发现 BP 1蛋白的加入增强了FGF在不同生物测定中的作用 系统.此外，BP 1 cDNA的表达在胚胎中是致死的， 转基因小鼠，并导致鸡胚血管渗漏。 BP mRNA在正常成年人或小鼠组织中低于检测，但在正常成年人或小鼠组织中低于检测。 可在小鼠胚胎肠道和皮肤中检测到。致癌物治疗上调 SCID中成年小鼠皮肤和致癌物处理的人皮肤异种移植物中的BP 1 小鼠BP 1在鳞状细胞癌（SCC）患者样本和细胞中表达 细胞系以及结肠癌细胞系和肿瘤样品中。相反，BP 2 似乎仅在黑色素瘤中上调，而在肿瘤中未检测到 表达BP 1的类型。 在人ME-180（SCC）和Ls 174 T（结肠）中观察到肿瘤生长的抑制 癌）细胞系，其中内源性BP 1水平已经用 BP 1靶向核酶。这一发现与BP 1的表达相结合， 提示BP 1对恶性肿瘤的限速作用。 SCC和结肠癌的表型。我们计划进行以下研究： 在目标1下，我们提出了一系列实验来定位BP / FGF蛋白/ 蛋白质相互作用来定义原子水平上的相互作用。下 目的2：我们将确定BPs和与硫酸乙酰肝素的相互作用对 FGF在细胞和组织中诱导的作用。在目标3下，我们将使用 核酶靶向BP mRNA，以评估BP 2的表达程度， 黑色素瘤的致瘤性表型的速率限制以及 肿瘤生长和转移BP 2和BP 1的作用最显著。下 目的4研究BP 1基因在转基因小鼠和鸡中表达的影响 胚胎组织选择性载体和条件性四环素依赖性 表达将用于避免BP 1表达的胚胎致死性。

DESCRIPTION (Applicant's Abstract): Fibroblast growth factors (FGF-1 or -2) are present at significant concentrations in most normal tissues in the adult. However, these FGFs are immobilized in an inactive state on the extracellular matrix and it is only poorly understood how they are solubilized and activated to reach their receptors. These growth factors can be mobilized by a previously identified secreted binding protein for FGF (BP1). We showed that the retinoid-regulated BP1 can enhance the activity of locally stored, immobilized FGFs and that expression of BP1 supports tumor growth and angiogenesis of FGF-2 positive non-tumorigenic cells. A recently identified novel BP2 showed similar biologic activities. In preliminary studies we found that addition of BP1 protein enhances FGF effects in different biological assay systems. Furthermore, expression of the BP1 cDNA was embryonically lethal in transgenic mice and caused vascular leakage in chicken embryos. BP mRNA was below detection in normal adult human or murine tissues but was detectable in murine embryonic gut and skin. Carcinogen treatment upregulates BP1 in adult mouse skin and in carcinogen-treated human skin xenografts in SCID mice. BP1 was expressed in squamous cell cancer (SCC) patient samples and cell lines as well as in colon cancer cell lines and tumor samples. In contrast, BP2 appears to be upregulated only in melanoma and was not detected in the tumor types expressing BP1. Inhibition of tumor growth was observed in human ME-180 (SCC) and Ls174T (colon cancer) cell lines in which the endogenous BP1 levels had been reduced with BP1-targeted ribozymes. This finding in conjunction with the expression of BP1 in human cancer samples suggests a rate-limiting role of BP1 for the malignant phenotype of SCC and colon cancer. We plan the following studies: Under aim 1 we propose a series of experiments to map the BP / FGF protein / protein interactions to define these interactions at the atomic level. Under aim 2 we will define the effects of BPs and interaction with heparansulfate on the effects induced by FGFs in cells and tissues. Under aim 3 we will use ribozyme-targeting of BP mRNA to assess to what extent expression of the BP2 is rate limiting for the tumorigenic phenotype of melanoma and to what stage of tumor growth and metastasis BP2 and BP1 contribute most significantly. Under aim 4 we will study the effect on BP1 expression in transgenic mice and chicken embryos. Tissue-selective vectors and conditional tetracycline-dependent expression will be used to circumvent embryonic lethality of BP1 expression.