The Role of LIN28B Signaling in Mediating the Oncogenic Phenotype in Neuroblas

 描述(申请人提供)：神经母细胞瘤说明了一种肿瘤组织类型可以包含的惊人的临床异质性，4S期患者接受自发性肿瘤消退，高风险患者表现出显著的死亡率。我们实验室最近发现Lin28b是神经母细胞瘤的致癌驱动因素，它是一种RNA结合蛋白，可以抑制let-7肿瘤抑制因子microRNA家族并直接与mRNAs结合。虽然它在新陈代谢和胚胎发育中的作用越来越为人所知，但它如何促进癌基因还不太清楚。我们已经证明，Lin28b可以驱动RAS相关蛋白RAN和极光蛋白A(AURKA)的表达，从而促进神经母细胞瘤的增殖。我们也有证据表明，Lin28b在神经母细胞瘤中促进无茎相关基因签名，增加了其在正常发育中促进自我更新的功能也可能起到致癌作用。这份有指导的职业发展建议将检验这样一个假设，即Lin28b与RAN和AURKA一起促进细胞增殖和自我更新，以驱动神经母细胞瘤的侵袭性。首先，我们将确定Lin28b促进自我更新的机制。其次，我们将剖析下游的Lin28b靶标RAN和AURKA对细胞增殖和自我更新的贡献。在最终目标中，我们将检验这样的假设，即Lin28b与mRNAs，包括RAN信号网的mRNAs的结合，提供了一种发挥其致癌功能的机制。这一K08奖将拓宽我的科学曲目，并为我在基础肿瘤学和转化性肿瘤学的职业生涯提供一个杰出的基础。首先，在目标1和目标2中，我将开发关于Lin28b/let-7在调节自我更新中的作用的新专业知识，这是癌症的一个标志，可能代表最终的治疗脆弱性，但这需要进一步研究。其次，我将掌握RNA-Seq及相关技术，使用它来生成和测试假设；在目标1中，我将使用RNA-Seq来揭示Lin28b用来调节自我更新的途径。最后，在目标3中，我将开发利用CLIP-Seq的专业知识，这是一种高通量技术，可以识别与RNA结合蛋白结合的RNA。这将进一步加深我们对受Lin28b影响的编码和非编码RNA的理解，并可能提高我们对其他RNA结合蛋白功能的理解。这项工作将在John Maris博士的杰出指导下完成，并由Anil Rustgi博士、Celest Simon博士、Andrei Thomas-Tikhonenko博士、Tom Look博士和Sharon Diskin博士组成的咨询委员会完成。总的来说，该委员会拥有癌症遗传学、RNA结合蛋白、microRNAs、干细胞生物学、生物信息学和疾病动物模型方面的专业知识。通过研究神经母细胞瘤的这一关键途径，我的目标是为其最终的治疗操作奠定基础，并产生新的见解，以加深我们对儿童和成人恶性肿瘤的理解。总而言之，这笔拨款的科学和教育部分都将加强我的科学基础，使我能够开始独立的研究生涯。

 DESCRIPTION (provided by applicant): Neuroblastoma illustrates the striking clinical heterogeneity that one tumor histotype can encompass, with stage 4S patients undergoing spontaneous tumor regression and high-risk patients exhibiting significant mortality. Our laboratory recently identified LIN28B, an RNA binding protein that inhibits the let-7 tumor suppressor microRNA family and binds mRNAs directly, as an oncogenic driver in neuroblastoma. While its roles in metabolism and embryonic development are becoming increasingly well-understood, how it promotes oncogene- sis is less clear. We have shown that LIN28B drives expression of the Ras-related protein RAN and Aurora kinase A (AURKA), to promote neuroblastoma proliferation. We also have evidence indicating that LIN28B promotes stemless related gene signatures in neuroblastoma, raising the possibility that its function in promoting self-renewal in normal development may also serve an oncogenic role. This Mentored Career Development proposal will test the hypothesis that LIN28B, in conjunction with RAN and AURKA, promotes cell proliferation and self-renewal to drive neuroblastoma aggression. First, we will determine the mechanisms by which LIN28B promotes self-renewal. Second, we will dissect the contribution of the downstream LIN28B targets RAN and AURKA to cell proliferation and self-renewal. In the final aim, we will test the hypothesis that LIN28B binding to mRNAs, including mRNAs of the RAN signaling network, provides one mechanism by which it exerts its oncogenic function. This K08 Award will broaden my scientific repertoire and provide an outstanding basis for a career in basic and translational oncology. First, in Aims 1 and 2, I will develop new expertise on the role of LIN28B/let-7 in mediating self-renewal, a hallmark of cancer that may represent an eventual therapeutic vulnerability but that re- quires further study. Second, I will master RNA-Seq and related technologies, using it to generate and test hypotheses; in Aim 1, I will use RNA-Seq to unveil the pathways LIN28B utilizes to regulate self-renewal. Finally, in Aim 3, I will develop expertise in utilizing CLIP-Seq, a high-throughput technology that identifies RNAs to which RNA binding proteins bind. This will further our understanding of coding and non-coding RNAs influenced by LIN28B and may improve our understanding of how other RNA binding proteins function. This work will be completed under the outstanding mentorship of Dr. John Maris and an Advisory Committee, comprised of Drs. Anil Rustgi, Celeste Simon, Andrei Thomas-Tikhonenko, Tom Look, and Sharon Diskin. Collectively, this committee possesses expertise in cancer genetics, RNA binding proteins, microRNAs, stem cell biology, bioinformatics, and animal models of disease. By studying this key pathway in neuroblastoma, I aim to lay the basis for its eventual therapeutic manipulation and generate novel insights to further our understanding of both pediatric and adult malignancies. In summary, both the scientific and educational components of this grant will strengthen my scientific foundation, allowing me to launch an independent research career.