MOLECULAR DETERMINANTS OF ANGIOGENSIS IN ORAL CANCER

描述：林根博士提出的职业发展计划由多个 通过具有新专业知识的实验室轮换，这将直接 适用于他提议的研究工作。时间范围从1 一个月到三个月，地点从全国各地到 同样的机构。在第一年里，他已经安排好了去看望他。 勒罗伊·胡德在华盛顿大学呆了一个月，学习了 微阵列技术。在第二年，他将在 洛约拉大学布莱恩·尼科洛夫博士的实验室将亲身实践 在开发逆转录病毒和腺病毒方面的经验 含有他感兴趣的基因。在第三年，他提议每3个月轮换一次 通过卢西奥·米勒博士在洛约拉的实验室了解如何 确定关键转录因子并进行启动子分析。在第四年，他 访问芝加哥大学Elaine Fuchs博士的实验室 3个月内产生转基因小鼠，在其中表达 转基因将在可诱导的内皮细胞特异性的控制下 推动者。最终轮换尚未确定，但仍将可用 以便以后考虑所需的最先进的技术。除了……之外 上述轮换，林根博士也计划参加和讲座 在洛约拉大学学习与研究伦理相关的课程。 所描述的研究项目与林根博士目前的R01相同 赠款，这是经过同行审查和资助的。具体目标是： 1)详细描述功能维甲酸受体的特性 Northern和Western在微血管内皮细胞中的表达 Blotts，并通过进行电迁移率位移分析来确定 表达受体的功能状态。 2)确定哪些表达的维甲酸受体负责 使微血管内皮细胞对诱导剂产生抵抗力 血管生成通过使用受体选择性配体，并通过引入 显性阴性、野生型或嵌合型维甲酸受体进入 微血管内皮细胞。 3)研究维甲酸对体内血管生成的抑制作用。 导入含有逆转录病毒载体的人微血管内皮细胞 将显性负性、野生型或嵌合型视黄酸受体注入小鼠体内 并观察它们与新形成的血管的结合情况。

DESCRIPTION: Dr Lingen's proposed career development plan consists of multiple rotations through laboratories with novel expertise which will be directly applicable to his proposed research endeavors. The time frames range from 1 month to 3 months and the locations vary from across the country to within the same institution. During the first year, he has made arrangements to visit Dr. Leroy Hood at the University of Washington for 1 month and learn the cDNA microarray technology. During the second year, he will work within the laboratory of Dr. Brian Nickoloff at Loyola University to gain hands-on experience in the development of both retroviruses and adenoviruses that contain his genes of interest. During year 3, he proposes a 3-month rotation through Dr. Lucio Miele's laboratory at Loyola to gain knowledge of how to identify key transcription factors and perform promoter analyses. In year 4, he visits the laboratory of Dr. Elaine Fuchs at the University of Chicago for a 3-month period to generate transgenic mice in which the expression of the transgenes will be under the control of an inducible endothelial cell specific promoter. The final rotation has not been determined, but will remain available for later consideration of state-of-the-art techniques needed. In addition to the above mentioned rotations, Dr. Lingen also plans to participate and lecture in courses related to research ethics at Loyola University. The described research project is identical to the Dr. Lingen's current R01 grant which has been peer-reviewed and funded. The specific aims are: 1) To characterize in detail the profile of functional retinoic acid receptors expressed in microvascular endothelial cells by performing Northern and Western blots, and by performing electromobility shift assays to determine the functional status of the expressed receptors. 2) To determine which of the expressed retinoic acid receptors are responsible for causing microvascular endothelial cells to become refractory to inducers of angiogenesis by employing receptor selective ligands, and by introducing either dominant negative, wild-type or chimeric retinoic acid receptors into microvascular endothelial cells. 3) To study retinoic acid-mediated inhibition of in vivo angiogenesis by introducing human microvascular endothelial cells containing a retrovirally driven dominant negative, wild-type or chimeric retinoic receptors into mice and observing their incorporation into newly forming vessels.