CITED1 nuclear localization and its role in Wilms' tumor pathogenesis

描述（由申请人提供）：独立之路奖概述了最初的两年指导培训计划，随后是三年的独立阶段，以加速Lovvorn博士向科学独立的过渡。在最初的指导阶段，Lovvorn博士建议在范德比尔特大学（Vanderbilt University）高产的主要研究者和受人尊敬的导师Mark de Caestecker博士的监督下，通过多学科项目扩展他在癌症和发育生物学领域的科学技能和知识。在系主任的支持下，Lovvorn博士目前正在实践一种工作努力公式，将他工作日时间的75%用于研究目的。本文详细的研究计划将为Lovvorn博士提供严格的基础生物化学技术培训，以测试Wilms肿瘤中转录辅助因子CITED1的核胞质穿梭的功能意义和调节机制。在de Caestecker博士的指导下，Lovvorn博士证明了CITED1的表达在人类和大鼠肾母细胞瘤中持续存在，CITED1通常在发育中的肾脏中活跃，但在成熟的肾脏中不存在。他还表明，CITED1的亚细胞定位从后肾间质的细胞质转移到恶性囊胚的细胞核，并且在原发人类Wilms肿瘤中，CITED1表达水平的升高与疾病的晚期相关。Lovvorn博士根据这些观察提出了两个假设：1 . CITED1核定位有助于Wilms的肿瘤发病；在Wilms的肿瘤发生过程中，CITED1的核细胞质穿梭异常。在他最初的目标中，Lovvorn博士将测试将CITED1限制在Wilms1肿瘤细胞的细胞核中的致病意义。他将在人类Wilms肿瘤细胞系WIT49中产生两种主要错误表达CITED1的细胞质或核部分的Wilms肿瘤细胞系，并将评估这些操作对Wilms肿瘤生物学的影响。在他的第二个目标中，Loworn博士将使用质谱法识别CITED1结合伙伴，这些结合伙伴对亚细胞区室具有特异性。这些数据将有助于确定CITED1修饰及其在Wilms肿瘤胚泡中调节核保留的机制。这些研究将提供一个框架，不仅了解改变的CITED1核胞质穿梭，以及它对Wilms肿瘤发病机制的贡献，而且为未来的独立研究提供必要的培训。

DESCRIPTION (provided by applicant): This Pathway to Independence Award outlines an initial two-year mentored training program followed by a three-year independent phase to expedite the transition to scientific independence for Dr. Lovvorn. In the initial mentored phase, Dr. Lovvorn proposes to expand his scientific skills and knowledge in the fields of Cancer and Developmental Biology through a multidisciplinary program under the supervision of Dr. Mark de Caestecker, a highly productive principle investigator and respected mentor at Vanderbilt University. With the endorsement of his Department Chair, Dr. Lovvorn currently is practicing a work effort formula that affords 75% of his weekday time for research purposes. The research plan detailed herein will provide Dr. Lovvorn with rigorous training in basic biochemistry techniques to test the functional significance of and mechanisms regulating nucleocytoplasmic shuttling of the transcriptional co-factor CITED1 in Wilms' tumor. Under Dr. de Caestecker's guidance, Dr. Lovvorn has shown that expression of,CITED1, normally active in the developing but absent in the mature kidney, persists in both human and rat Wilms' tumors. He also has shown that the subcellular localization of CITED1 shifts from the cytoplasm in metanephric mesenchyme to the nucleus of malignant blastema, and that increasing levels of CITED1 expression in primary human Wilms' tumors correlate with advanced stages of disease. Dr. Lovvorn proposes two hypotheses from these observations: 1. CITED1 nuclear localization facilitates Wilms' tumor pathogenesis, and 2. Nucleocytoplasmic shuttling of CITED1 is dysregulated in Wilms' tumorigenesis. In his initial aim, Dr. Lovvorn will test the pathogenic significance of restricting CITED1 to the nucleus of Wilms1 tumor cells. He will generate two Wilms' tumor cell lines that misexpress either a predominantly cytosolic or nuclear fraction of CITED1 in the human Wilms' tumor cell line, WIT49, and will assess the effects of these manipulations on Wilms' tumor biology. In his second aim, Dr. Loworn, using mass spectrometry, will identify CITED1 binding partners that are specific to either subcellular compartment. These data will facilitate identification of CITED1 modifications and mechanisms regulating its nuclear retention in Wilms' tumor blastema. These studies will provide a framework not only to understand altered CITED1 nucleocytoplasmic shuttling in, and its contributions to, Wilms' tumor pathogenesis, but also the necessary training to pursue future independent investigation.