CITED1 nuclear localization and its role in Wilms' tumor pathogenesis

该独立之路奖概述了最初的为期两年的指导培训计划，然后是为期三年的独立阶段，以加快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富有成效的主要研究者和受人尊敬的导师Mark de Caestecker博士的监督下，通过多学科计划扩大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，该公式提供了75%的工作日时间用于研究目的。本文详细介绍的研究计划将为Lovvorn博士提供基础生物化学技术的严格培训，以测试Wilms肿瘤中转录辅因子CITED 1的核质穿梭的功能意义和调节机制。在de Caestecker博士的指导下，Lovvorn博士已经证明，CITED 1的表达在发育中通常是活跃的，但在成熟的肾脏中不存在，在人类和大鼠的Wilms肿瘤中持续存在。他还表明，CITED 1的亚细胞定位从后肾间充质中的细胞质转移到恶性胚基的细胞核，并且原发性人肾母细胞瘤中CITED 1表达水平的增加与疾病的晚期相关。Lovvorn博士从这些观察中提出了两个假设：1。CITED1核定位促进Wilms'肿瘤发病机制，和2。CITED 1的核质穿梭在Wilms肿瘤发生中失调。在他的最初目标中，Lovvorn博士将测试将CITED 1限制在Wilms 1肿瘤细胞核中的致病意义。他将产生两个Wilms肿瘤细胞系，其在人Wilms肿瘤细胞系WIT49中错误表达CITED 1的主要胞质或核部分，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博士将使用质谱法确定CITED1结合伴侣，这些伴侣对任一亚细胞区室都是特异性的。这些数据将有助于鉴定CITED 1修饰和调节其在肾母细胞瘤胚基中的核保留的机制。这些研究将提供一个框架，不仅要了解改变CITED1核质穿梭，其贡献，肾母细胞瘤的发病机制，但也是必要的培训，以追求未来的独立调查。

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.