T(15;19) in Aggressive Pediatric Carcinoma

描述(由申请人提供)：应聘者是一名病理学家 在过去的一年里全职从事研究，并获得了大量的 具有分子细胞遗传学和基因定位方法的经验。通过这件事 在工作中，他发现了一种以染色体为靶点的融合癌基因 易位，t(15；19)，在一种极具侵袭性的癌症中。 这份申请概述了一个计划，以提供对癌症生物学的广泛接触。 研究方法并阐明与之相关的致癌机制 T(15；19)。目标是：(1)学习概念性和理论性技能 癌蛋白的分子遗传学和功能分析方法；以及 (2)鉴定(15；19)相关融合癌蛋白并揭示其功能 这种蛋白质的性质。压倒一切的目标是确定候选人 作为一名独立资助的内科科学家，他可以贡献原创 癌症生物学和临床医学的发现和概念。这个 环境非常适合这一目标。S导师实验室已经 在分子遗传学和功能分析方面有丰富的专业知识 致癌基因。此外，候选人？S顾问委员会有广泛的 使用体外和小鼠模型的癌症生物学经验。导师和 监督委员会将促进科学互动和 合作，并将确保密集的课程，包括实验室 哈佛医学院内外的工作和教学课程 为了达到在癌症生物学方面有广泛经验的目标。 研究部分集中在一种毁灭性的癌症上， 易位t(15；19)(q13；p13.1)。这种独特的癌症已经完全 对多药化疗和放射治疗无反应，所有 患者(儿童和年轻人)在9个月内死亡 诊断。候选人通过以下方式本地化了易位断点 细胞遗传学、荧光原位杂交、Southern blotting和 RT-PCR方法。他的初步研究最近确定了核聚变 癌基因，由编码细胞的19号染色体配对癌基因组成 与15号染色体融合的增殖相关溴域蛋白 伙伴癌基因编码一种新的富含脯氨酸的蛋白质。未来的研究将 聚焦三个目标。首先，将全长t(15；19)融合癌基因 以快速扩增cDNA端为特征的，如有必要，通过 从t(15；19)癌细胞系中筛选cDNA文库。第二，他会 建立评估t(15；19)癌蛋白的体外模型 转化活动和转化机制。这些研究将解决 假设t(15；19)癌蛋白通过解除对T(15；19)的调节作用。 一种或两种其组成的天然蛋白质。第三，他将决定 利用FISH和RT-PCR检测t(15；19)在临床侵袭性疾病中的频率 来自儿童患者的癌症。这些研究也将揭示出 T(15；19)+癌的原发部位。

DESCRIPTION (provided by applicant): The candidate is a pathologist who has performed research full-time over the past year and has acquired substantial experience in molecular cytogenetic and gene mapping methods. Through this work, he has identified a fusion oncogene targeted by a chromosomal translocation, t(15;19), in an extraordinarily aggressive form of carcinoma. This application outlines a plan to provide a broad exposure to cancer biology research methods and to elucidate the oncogenic mechanisms associated with t(15;19). The aims are to: (1) learn skills, conceptual and theoretical approaches to molecular genetics and functional analysis of oncoproteins; and (2) identify the (15;19)-associated fusion oncoprotein and reveal functional properties of this protein. The overriding aim is to establish the candidate as an independently funded physician-scientist who can contribute original findings and concepts to cancer biology and clinical medicine. The environment is well suited to this goal. The mentor?s laboratory has substantial expertise in molecular genetics and the functional analysis of oncogenes. In addition, the candidate?s advisory committee has broad experience in cancer biology using in vitro and murine models. The mentor and t h e a dvisory committee will facilitate scientific interactions and collaborations, and will ensure an intensive curriculum, including laboratory work and didactic courses, both within and outside of the Harvard Medical Area, in order to meet the aim of a broad experience in cancer biology. The research component focuses on a devastating form of carcinoma with translocation t(15;19)(q13;p13.1). This unique carcinoma has been completely unresponsive to multi-agent chemotherapy and radiation therapy, and all patients (children and young adults) have succumbed within 9 months of diagnosis. The candidate has localized the translocation breakpoints by cytogenetic, fluorescence in situ hybridization (FISH), Southern blotting, and RT-PCR methods. His preliminary studies have recently identified the fusion oncogene, comprised of a chromosome 19 partner oncogene encoding a cell proliferation-related bromodomain protein, fused with the chromosome 15 partner oncogene encoding a novel proline-rich protein. Future studies will focus on three aims. First, the full length t(15;19) fusion oncogene will be characterized by rapid amplification of cDNA ends and, if necessary, by screening cDNA libraries from a t(15;19) cancer cell line. Second, he will establish in vitro models with which to evaluate the t(15;19) oncoprotein transforming activity and transforming mechanisms. These studies will address the hypothesis that the t(15;19) oncoprotein acts by deregulating function of one or both of its component native proteins. Third, he will determine the frequency of t(15;19), using FISH and RT-PCR assays, in clinically aggressive carcinomas from pediatric patients. These studies will also reveal the range of primary sites from which t(15;19)+ carcinomas arise.