The role of AF10 in normal and leukemic hematopoiesis

项目概述：本项目的总体目标是提供新的见解的机制， 血液恶性肿瘤中的表观遗传失调可能导致改善的治疗效果 战略布局携带AF 10转录因子的染色体重排的白血病由以下标记： 染色质1-3的特异性扰动，从而为表观遗传学的研究提供了一个很好的模型。 肿瘤发生的失调。我们假设AF 10白血病的表观遗传失调是由于 AF 10 N端PHD结构域的缺失和C端八肽基序-亮氨酸的保留 拉链（OM-LZ）结构域在致癌AF 10融合。我们的初步观察表明， AF 10融合癌基因可通过抑制AF 10 OM-LZ结构域的活性或 或者，通过恢复致癌性AF 10融合体中的PHD锌指结构域。这些观察结果仍然有效 对AF 10白血病有很好的治疗前景。在具体目标1中，我们建议确定 使用已建立的白血病和基因敲除小鼠模型， 吞吐量接近。这些研究的临床相关性通过AF 10参与了 多种血液恶性肿瘤中的复发性染色体易位4-8.此外，若干 其他癌症与AF 10白血病具有共同的特征，特别是涉及PHD结构域9-12或 组蛋白甲基转移酶DOT 1 L 13以及发育关键HOX基因的失调14，15。 因此，拟议的研究可能会产生有价值的见解HOX基因的共同机制， 癌症的失调我们的初步结果支持AF 10在哺乳动物HOX调控中的作用 基因.这种功能的失调似乎是AF 10白血病肿瘤发生的关键事件。因此，我们认为， 为了深入了解AF 10在正常和白血病造血中的作用，我们建议确定AF 10在正常和白血病造血中的作用。 使用生物化学和基因靶向方法在正常和白血病造血中的AF 10具体地说， 我们计划表征AF 10的转录活性，并建立条件性敲除小鼠， 特异性目标2中的造血特异性缺失AF 10。先前的研究支持AF 10在 HOX基因的调节1，2，16，17。建立AF 10敲除小鼠将能够评估AF 10 功能，包括HOX基因的潜在调节，在正常和恶性肿瘤中发挥重要作用， 造血拟议的项目将在丰富的学术机构环境中进行，并将 由著名科学家组成的指导委员会指导。该建议得到了一个强有力的支持。 培训部分，包括表观遗传学和血液病理学的教学和实践指导，培训 在负责任地进行研究，并获得指导能力，成功地过渡到一个 独立调查员

PROJECT SUMMARY: The overall goal of this project is to offer novel insights into the mechanisms of epigenetic deregulation in hematological malignancies that may lead to improved therapeutic strategies. Leukemias bearing chromosomal rearrangements of the AF10 transcription factor are marked by specific perturbations of chromatin 1-3, thereby providing an excellent model for the study of epigenetic deregulation in oncogenesis. We hypothesize that epigenetic dysregulation in the AF10 leukemias results from the loss of the N-terminal PHD domains of AF10 and the retention of the C-terminal octapeptide motif-leucine zipper (OM-LZ) domain in oncogenic AF10 fusions. Our preliminary observations show that transformation by AF10 fusion oncogenes can be suppressed by inhibiting the activity of the AF10 OM-LZ domain or alternatively, by restoring the PHD zinc finger domain in oncogenic AF10 fusions. These observations hold great therapeutic promise for the AF10 leukemias. In Specific Aim 1, we propose to identify mechanisms that lead to tumor suppression using established models of leukemia and knockout mice, followed by high throughput approaches. The clinical relevance of these studies is borne out by the fact that AF10 is involved in recurrent chromosomal translocations in a wide variety of hematological malignancies 4-8. Moreover, several other cancers share features common to the AF10 leukemias, notably - involvement of PHD domains 9-12 or the histone methyltransferase DOT1L 13 together with deregulation of the developmentally critical HOX genes 14, 15. The proposed studies may therefore yield valuable insights into common mechanisms of HOX gene deregulation in cancer. Our preliminary results support a role for AF10 in the regulation of mammalian HOX genes. Deregulation of this function seems to be a critical event in oncogenesis of AF10 leukemias. Therefore, to gain insights into the role of AF10 in normal and leukemic hematopoiesis, we propose to identify the role of AF10 in normal and leukemic hematopoiesis using biochemical and gene-targeting approaches. Specifically, we plan to characterize the transcriptional activity of AF10 and establish conditional knockout mice with hematopoietic specific deletions of AF10 in Specific Aim 2. Prior studies support a role for AF10 in the regulation of HOX genes 1, 2, 16, 17. Establishment of AF10 knockout mice will enable assessment of AF10 function including potential regulation of HOX genes that have important roles in normal and malignant hematopoiesis. The proposed project will be carried out in a rich academic institutional environment, and will be guided by a mentoring committee composed of renowned scientists. The proposal is supported by a strong training component that includes didactic and practical instruction in epigenetics and hematopathology, training in the responsible conduct of research, and acquisition of mentoring abilities for successfully transitioning to an independent investigator.