The role of the histone chaperone Chaf1b in sustaining the Hoxa9-driven AML differentiation block

组蛋白伴侣 Chaf1b 在维持 Hoxa9 驱动的 AML 分化阻断中的作用

基本信息

批准号：
9295511
负责人：
Mario Andres Blanco
金额：
$ 19.2万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9295511
关键词：
ATAC-seq Acute Myelocytic Leukemia Affect Animal Model Automobile Driving Bachelor&apos s Degree Biochemical Biochemistry Blast Cell Blood Boston Breast cancer metastasis Cancer Biology Cancer Etiology Cell Cycle Arrest Cell Differentiation process Cell physiology Cells Cessation of life ChIP-seq Chromatin Collaborations Complex Computational Biology Consultations DNA DNA Modification Process Data Development Differentiated Gene Differentiation Therapy Doctor of Philosophy Epigenetic Process Faculty Fellowship Foundations Funding Gene Expression Genetic Genetic Screening Genetic Transcription Genomics Goals Grant Hematologic Neoplasms Hematopoiesis Hematopoietic Neoplasms Hematopoietic stem cells Histones Interview Journals Knockout Mice Laboratories MLL-AF9 Malignant Neoplasms Malignant neoplasm of brain Mediating Methodology Modeling Molecular Molecular Biology Molecular Chaperones Mus Myelogenous Nature Neoadjuvant Therapy Nucleosomes Paper Pathology Patients Pediatric Hospitals Pennsylvania Phase Physiological Plant Roots Positioning Attribute Postdoctoral Fellow Process Proteins Publishing RNA RNA immunoprecipitation sequencing Relapse Reporting Research Research Personnel Role Schools Scientist Societies Solid Neoplasm Stimulus Testing Therapeutic Training Transgenes Tretinoin United States National Institutes of Health Universities Untranslated RNA Vision Visit Work Writing Yang anticancer research base blocking factor burden of illness career cell type cellular pathology chemotherapy chromatin remodeling clinically relevant design epigenetic regulation epigenomics in vivo interest leukemia/lymphoma medical schools member mouse model novel programs recruit response screening small hairpin RNA stemness tenure track therapeutic candidate therapeutic target transcription factor transcriptome sequencing transcriptomics

项目摘要

Project Summary/Abstract I received my undergraduate degree from Cornell University, my PhD in molecular biology from Princeton University, and am currently a postdoctoral researcher and Special Fellow of the Leukemia and Lymphoma Society at Harvard Medical School and Boston Children's Hospital. After studying the molecular basis of breast cancer metastasis in graduate school, I transitioned to the field of epigenetics in the laboratory of Yang Shi for my postdoc. I have received extensive experimental and scholarly training in my postdoc. I learned the rigors of basic chromatin biochemistry in my first postdoctoral project, and recently published a co-first author paper based on this work. This study was featured as the cover article in the journal Cell and reported the identification of a novel metazoan epigenetic DNA modification, N6-methyladenine (6mA). My scholarly training has involved extensive scientific writing and has enabled me to write three successful postdoc fellowship applications (from the NIH, ACS, and LLS), two successful grants for Dr. Shi based on my work (from the Samuel Waxman Cancer Research Foundation and the Harvard Epigenetics Initiative), and one recently published review paper as a co-author in the journal Nature Reviews Molecular Biology. My current postdoctoral research merges my background in cancer biology with my recent training in chromatin biochemistry and focuses on the epigenetic basis of the differentiation block that characterizes acute myeloid leukemia (AML). AML is the most lethal hematological malignancy and is the cause of more than 10,000 annual US death. AML is typically treated by chemotherapy, though patients often relapse and have limited therapeutic options. The promyelocytic subtype of AML can be cured by “differentiation therapy” – induction of differentiation and inhibition of proliferation with retinoic acid – but this approach has been ineffective in other AML subtypes. Recent work has suggested that this block to non-APL AML differentiation is epigenetic in nature. Stable – yet reversible – chromatin alterations are thought to disable myeloid differentiation gene expression programs in these cells. I have recently established a robust genetic screening approach to uncover epigenetic regulators of non-APL AML differentiation programs. These screens have identified the Chaf1b subunit of the Caf-1 histone remodeling complex as a critical regulator of the Hoxa9- driven AML differentiation block. The overall goal of the work proposed in this application is to determine the molecular, chromatin-based mechanisms by which Chaf1b functions to regulate AML cell differentiation. The goal of my first Aim in this proposal is to use biochemical approaches to identify the proteins and long non- coding RNAs (lncRNAs) that associate with Chaf1b and the greater Caf-1 complex in AML cells. I will then focus on determining which of these interacting proteins and RNAs are most critical in recruiting Chaf1b to its target genomic loci and help sustain its activity as a differentiation-blocking protein. In Aim 2 I will use extensive epigenomic profiling approaches to identify the chromatin remodeling and transcriptomic processes that are altered as cells differentiate in response to inhibition of Chaf1b. I will then compare these epigenomic data to those obtained from my recent profiling of the normal, physiological myeloid differentiation program to determine whether loss of Chaf1b induces differentiation through typical or atypical mechanisms. Finally, in Aim 3 I will determine the potential of Chaf1b as a candidate therapeutic target by testing whether its inhibition induces differentiation and reduces disease burden or affects normal hematopoiesis in mouse AML models. While I am currently a postdoc, the proposed research is to be carried out as an independent investigator, which is my immediate career goal. The initial phase of preparing for this transition has been very promising. I have recently had interviews for tenure-track faculty positions at the University of Pennsylvania, Princeton University, and the Mount Sinai School of Medicine. Mt. Sinai has recently invited me for a second visit and I am awaiting responses from UPenn and Princeton. To help facilitate this transition, I will have collaborative and consultative support of field leaders in hematopoiesis (Dr. David Scadden), AML (Dr. Kimberly Stegmaier), and lncRNA/computational biology (Dr. John Rinn). These scientists are committed to guiding me through the transition process and helping me start my independent career in the strongest possible position. Once independent, my research will focus on epigenetic regulation of AML cell differentiation. However, my long term vision is to bring the concept of targeting cell fate decisions from blood lineage cancers to selected solid tumors of interest. “Differentiation therapy” has been underexplored in solid tumors, but the importance of stemness and de-differentiation programs has been clearly implicated in certain malignancies such as brain cancer. Therefore, toward the end of the proposed funding period, I will aim to establish differentiation-based screening methodology for selected brain cancers and will work on characterizing the molecular mechanisms of screen hits using a combination of genetic, biochemical, epigenomic, and animal models. I believe that this direction will lead to conceptual and therapeutic advances and will have direct clinical relevance.

项目摘要/摘要我获得了康奈尔大学的本科学位，我的分子生物学博士学位大学，目前是博士后研究员和白血病和淋巴瘤的特别研究员哈佛医学院和波士顿儿童医院的协会。研究了分子基础之后我在研究生院的乳腺癌转移，在Yang实验室过渡到表观遗传学领域 Shi为我的博士后。我在博士后接受了广泛的实验和科学培训。我学到了我的第一个博士后项目中的基本染色质生物化学的严格基于这项工作的纸。这项研究以详情的Cell杂志的封面文章为特征，并报道了鉴定新型后生表观遗传学DNA修饰，N6-甲基趋化（6mA）。我的科学培训涉及广泛的科学写作，使我能够写三个成功的博士后奖学金申请（来自NIH，ACS和LLS），根据我的工作，Shi博士的两项成功赠款（摘自Samuel Waxman Cancer Research Foundation和Harvard表观遗传学倡议），一项最近发表的评论论文是《自然评论分子生物学》杂志的合着者。我目前的博士后研究将我的癌症生物学背景与我最近的培训相结合染色质生物化学，并重点介绍分化块的表观遗传基础，该基础表征了急性髓样白血病（AML）。 AML是最致命的血液系统恶性肿瘤，其原因超过 10,000年美国死亡。 AML通常通过化学疗法治疗，尽管患者经常中继有限的治疗选择。 AML的临时细胞亚型可以通过“分化疗法”来治愈 - 诱导分化和用视黄酸抑制增殖的抑制作用 - 但是这种方法已经是在其他AML亚型中无效。最近的工作表明，这个非APL AML分化的障碍本质上是表观遗传学的。稳定的，但可逆 - 染色质的改变被认为禁用髓样这些细胞中的分化基因表达程序。我最近建立了强大的基因筛查发现非APL AML分化程序的表观遗传调节剂的方法。这些屏幕有 CAF-1 Hisstone重塑复合体的CHAF1B亚基是Hoxa9-的关键调节剂驱动的AML分化块。本应用程序提出的工作的总体目标是确定 CHAF1B通过其调节AML细胞分化的基于分子的染色质机制。我在该提案中的第一个目标的目的是使用生化方法来识别蛋白质和长期非 - 与AML细胞中CHAF1B和更大CAF-1复合物相关的编码RNA（LNCRNA）。然后我会专注于确定这些相互作用的蛋白质和RNA中的哪一种对于招募CHAF1B至关重要靶基因组基因座并有助于维持其作为分化阻滞蛋白的活性。在AIM 2中，我会使用广泛的表观基因组分析方法，以识别染色质重塑和转录过程随着细胞响应CHAF1B的响应，随着细胞的分化而改变的。然后，我将比较这些表观基因组从我最近对正常的物理髓样分化程序进行分析到的数据确定CHAF1B的损失是通过典型或非典型机制诱导分化的。最后，在 AIM 3我将通过测试其抑制作用来确定CHAF1B作为候选治疗目标的潜力在小鼠AML模型中诱导分化并减少疾病伯恩或影响正常造血。虽然我目前是博士后，但拟议的研究应作为独立研究者进行，这是我的直接职业目标。准备这种过渡的初始阶段是非常有希望的。我最近在宾夕法尼亚大学的普林斯顿大学接受了终身教师职位的采访大学和西奈山医学院。西奈山最近邀请我进行第二次访问，我正在等待Upenn和Princeton的回应。为了帮助促进这一过渡，我将进行协作和对造血症（David Scadden博士），AML（Kimberly Stegmaier博士）和 LNCRNA/计算生物学（John Rinn博士）。这些科学家致力于指导我过渡过程，并帮助我在强大的职位上开始自己的独立职业。一旦独立，我的研究将集中于AML细胞分化的表观遗传调节。但是，我的长期视觉是将靶向细胞命运决策的概念从血统癌症到选定的概念实体瘤感兴趣。 “分化疗法”在实体瘤中尚未得到充实，但是在某些恶性肿瘤（如大脑）癌症。因此，在拟议的融资期结束时，我将旨在建立基于差异化的选定脑癌的筛查方法，并将致力于表征分子机制使用遗传，生化，表观基因组和动物模型的筛选命中。我相信这个方向将导致概念和治疗的进步，并具有直接的临床相关性。