Elucidating transcription regulation by epigenetics in neuroblastoma

项目摘要/摘要 需要对高危神经母细胞瘤儿童进行更有效的治疗，因为5年生存率仍然较低 而不是50%。目前的风险分类无法预测哪些高危患者可能对 预先治疗或复发。因此，对分子机制的更好理解对 神经母细胞瘤的生长和存活是完善风险分层和开发新的 治疗学。已知在实体肿瘤中存在严重缺氧区，影响许多细胞。 治疗途径和对常用疗法的抵抗力增加。我之前发现了转录的 低氧诱导神经母细胞瘤恶性行为的通路及其可能的治疗作用 目标。低氧还会导致表观遗传变化，包括TET1的下调，TET1是一种转化为 5-甲基胞嘧啶(5mC)至5-羟甲基胞嘧啶(5hmC)。相比之下，最近的研究表明，5hmC，a 在神经母细胞瘤中，被激活的RNA转录的标志物在低氧条件下增加。我的初步数据显示 这种独特的缺氧反应存在于致癌神经母细胞瘤细胞系中，而不存在于非致癌神经母细胞瘤细胞系中 细胞系。我假设在低氧中上调TET1和增加5hmC水平会诱导转录 推动临床侵袭性神经母细胞瘤肿瘤生长的变化可用于对患者进行风险分层，并可能 最终导致新的治疗靶点的发现。我的提案的具体目标1将产生和 根据5hmC调节的转录图谱验证神经母细胞瘤风险分类器并识别潜在的 治疗靶点受这些缺氧诱导的表观遗传修饰的调节。在具体目标2中，我将 对几个不同表型的神经母细胞瘤进行深入的表观遗传学和转录分析 表现出对低氧的异质性TET1反应的细胞系。然后，我将能够测试 低氧诱导的表观遗传调控的基因和途径，以确定新的治疗策略。这些 实验将为进一步的体内研究和临床研究建立临床前数据，这对未来的R01水平研究至关重要。 IES正在寻找抑制神经母细胞瘤中表观遗传驱动的缺氧反应的药物。我提议四个- 一年的职业发展计划，以确立我作为神经母细胞瘤癌症独立研究员的地位 基因组学。我的发展计划由教学、机构资源和有监督的研究组成 数据聚集、统计方法和癌症表观遗传调控的算法方法。我有过 组建了一支国际公认的生物信息学一流专家团队(罗伯特·格罗斯曼博士)， 表观遗传学(Lucy Godley，医学博士)、神经母细胞瘤(Susan Cohn，医学博士)和统计遗传学(Barbara 陌生人，博士)作为导师来监督这次培训。具有职业发展计划，该计划利用 在芝加哥大学丰富的学术环境下，我将发展成为一名独立的医生-- 能够在治疗学、生物信息学和神经母细胞瘤之间架起桥梁的科学家。

PROJECT SUMMARY/ABSTRACT More effective therapy is needed for children with high-risk neuroblastoma as 5-year survival rates remain less than 50%. The current risk classification is unable to predict which high-risk patients are likely to respond to upfront treatment or relapse. Thus, an improved understanding of the molecular mechanisms important to the growth and survival of neuroblastoma is critical to refining risk stratification and the development of novel therapeutics. Regions of severe hypoxia are known to be present in solid tumors, affecting numerous cellular pathways and increasing resistance to common therapeutics. I have previously identified transcriptional pathways induced by hypoxia that promote malignant behavior in neuroblastoma and are possible therapeutic targets. Hypoxia also induces epigenetic changes including down-regulation of TET1, an enzyme that converts 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC). In contrast, recent studies have shown 5hmC, a marker of activated RNA transcription, is increased by hypoxia in neuroblastoma. My preliminary data show that this unique hypoxia response is present in tumorigenic neuroblastoma cell lines but not in non-tumorigenic cell lines. I hypothesize that up-regulated TET1 and increased 5hmC levels in hypoxia induce transcriptional changes driving clinically aggressive neuroblastoma tumor growth can be used to risk stratify patients and may ultimately lead to the discovery of new therapeutic targets. Specific Aim 1 of my proposal will generate and validate a neuroblasotma risk classifier based on 5hmC-regulated transcriptional profiles and identify potential therapeutic targets regulated by the these hypoxia-induced epigenetic modifications. In Specific Aim 2, I will perform an in depth epigenetic and transcriptional analysis of several phenotypically distinct neuroblastoma cell lines that exhibit heterogeneous TET1 responses to hypoxia. I will then be able to test the functional role of hypoxia-induced, epigenetically regulated genes and pathways to identify novel therapeutic strategies. These experiments will establish preclinical data for further in vivo and clinical studies critical for future R01-level stud- ies to identify drugs that inhibit the epigenetically driven hypoxia-response in neuroblastoma. I propose a four- year career development plan to establish myself as an independent investigator in neuroblastoma cancer genomics. My development plan is comprised of didactics, institutional resources, and supervised research in algorithmic approaches to data clustering, statistical methods, and the epigenetic regulation of cancer. I have assembled a first-class team internationally recognized experts in bioinformatics (Robert Grossman, PhD), epigenetics (Lucy Godley, MD, PhD), neuroblastoma (Susan Cohn, MD), and statistical genetics (Barbara Stranger, PhD) as mentors to oversee this training. With a career development plan which takes advantage of the rich academic environment at the University of Chicago, I will develop into an independent physician- scientist who can bridge the gulf between therapeutics, bioinformatics, and neuroblastoma.