Epigenetic Markings in Developing and Diseased Prefrontal Neurons

发育中和患病前额神经元的表观遗传标记

• 批准号: 7802630
• 负责人: Schahram Akbarian
• 金额: $ 50万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802630
• 关键词: Ablation; Adolescent; Affect; Age; Animals; Antipsychotic Agents; Anxiety; Applications Grants; Area; Autopsy; Award; Axon; Back; Behavioral; Biological Assay; Brain; Brain region; Cell Nucleus; Cells; Cerebral cortex; Cessation of life; Chemicals; Chromatin; Complement; DNA Sequence; DNA-Directed RNA Polymerase; Dependovirus; Development; Diagnosis; Disease; Disease model; Epigenetic Process; Etiology; Fiber; Fluorescence; Functional disorder; Gene Activation; Gene Expression; Genes; Genetic; Genetic Code; Genetic Transcription; Grant; Heterogeneity; Histone H3; Histone-Lysine N-Methyltransferase; Histones; Human; Human Development; Individual; Laboratories; Life; Longevity; Lysine; Maps; Mental disorders; Methodology; Methylation; Methyltransferase; Modeling; Modification; Molecular; Monitor; Mus; Mutant Strains Mice; Neurobiology; Neurons; Neurosciences; Occupations; Patients; Pattern; Play; Prefrontal Cortex; RNA; Regulation; Research; Resolution; Rodent Model; Role; Schizophrenia; Short-Term Memory; Site; Sorting - Cell Movement; Specimen; Synapses; Techniques; Testing; Time; Tissues; Transgenes; Ultracentrifugation; Work; age related; base; behavior test; brain tissue; chromatin immunoprecipitation; chromatin remodeling; design; epigenomics; genome-wide; innovation; insight; leukemia; mature animal; mutant; myelination; neuronal circuitry; neurotrophic factor; postnatal; prenatal; prepulse inhibition; promoter; recombinase; research study; theories

DESCRIPTION (provided by applicant): Schizophrenia is defined by a lack of a straightforward genetic cause for a very large majority of affected individuals. The prefrontal cortex (PFC), among other brain regions, is thought to be frequently in subjects in schizophrenia, as reflected by its functional hypoactivity and dysregulated expression for a diverse set of genes. The underlying molecular mechanisms remain unknown but there is evidence that the prolonged maturation of the PFC, extending into or even beyond the second decade of life, plays a crucial role for normal human development and the neurobiology of schizophrenia. Recently, we presented the first evidence that a subset of epigenetic markings, including trimethylated histone H3-lysine 4 at sites of gene promoters, is involved in the dynamic regulation of PFC chromatin during throughout pre- and postnatal development, and may be altered in some cases with schizophrenia. This challenge grant proposal is "shovel ready" and create new jobs as well as retain existing ones. Specifically, we will combine two of the most innovative approaches in the field of neurosciences as it pertains to epigenetics, by selectively sorting neuronal chromatin followed by massively parallel sequencing of immunprecipitates (ChIP-seq) to obtain insight into the epigenomic landscape of prefrontal neurons. We will profile developmental and disease-related changes in histone methylation markings associated with active promoters (trimethyl-H3K4) or transcription (trimethyl-H3K36) selectively in PFC neurons. Furthermore, we will study in conditional mutant mice the role of a candidate histone lysine methyltransferase, Mll1 (mixed-lineage leukemia 1) for normal and diseased prefrontal development, by expressing Cre recombinase selectively in weanling PFC and then monitor behavioral and molecular alterations in the adult animal. The experiments proposed here, which are designed to be accomplished within the challenge grant's award period of 24 months, will clarify, for the first time, whether or not chromatin of human PFC neurons is developmentally regulated and altered in disease. The answer to this question on a genome-wide level will be critical in order to find out whether or not chromatin-based mechanisms are part of a final common pathophysiology underlying prefrontal dysfunction in the substantial portion of subjects diagnosed with schizophrenia. PUBLICH HEALTH RELEVANCE: For the majority of patients diagnosed with schizophrenia, no straightforward genetic cause has been identified. One of the important theories about schizophrenia implies that in some brain regions, such as the "prefrontal cortex", a number of genes are not switched on properly during normal development, as they are in healthy subjects. However, until now the molecular techniques to study these phenomena have been lacking. This challenge grant proposal is "shovel ready", will create new jobs and retain existing ones. It is also based on extremely innovative techniques that were recently developed in our laboratories. We were able, for the first time, to selectively isolate chromosomal materials and chromatin from the nerve cells of the human brain obtained postmortem, and study "epigenetic markings" (basically, chemical modifications that regulate gene expression and function without altering the genetic code) on a genome-wide level. In this proposal, we plan to examine epigenetic gene activation patterns during normal prefrontal cortex development and search for potential alterations in schizophrenia and in mutant mice designed to model the disease on a molecular level. This research should clarify whether or not epigenetic markings are dynamically regulated in human neuronal chromatin , and if major psychiatric diseases such as schizophrenia are associated with changes in the neuronal epigenome.

描述（由申请人提供）：精神分裂症的定义是一个缺乏一个直接的遗传原因，为一个非常大的多数受影响的个人。前额叶皮层（PFC），在其他大脑区域中，被认为是精神分裂症受试者中常见的，这反映在其功能减退和多种基因表达失调。潜在的分子机制仍然未知，但有证据表明，PFC的长期成熟，延伸到甚至超过生命的第二个十年，对人类正常发育和精神分裂症的神经生物学起着至关重要的作用。最近，我们提出了第一个证据表明，一个子集的表观遗传标记，包括三甲基化组蛋白H3-赖氨酸4在基因启动子的网站，参与PFC染色质的动态调节在整个产前和产后的发展，并可能会改变在某些情况下与精神分裂症。这项挑战性的赠款提案是“铲准备”，创造新的就业机会，以及保留现有的。具体来说，我们将结合联合收割机在神经科学领域的两个最具创新性的方法，因为它涉及表观遗传学，通过选择性地分选神经元染色质，然后通过免疫沉淀物的大规模平行测序（ChIP-seq），以深入了解前额叶神经元的表观基因组景观。我们将在PFC神经元中选择性地描述与活性启动子（三甲基-H3 K4）或转录（三甲基-H3 K36）相关的组蛋白甲基化标记的发育和疾病相关变化。此外，我们将研究在条件突变小鼠的候选组蛋白赖氨酸甲基转移酶，MLL 1（混合谱系白血病1）的正常和患病的前额叶发育的作用，通过表达Cre重组酶选择性断奶PFC，然后监测成年动物的行为和分子改变。这里提出的实验，其目的是在24个月的挑战补助金的奖励期内完成，将澄清，第一次，人类PFC神经元的染色质是否发育调节和改变的疾病。在全基因组水平上回答这个问题将是至关重要的，以找出是否染色质为基础的机制是一个最终的共同病理生理学的一部分，潜在的前额叶功能障碍的大部分被诊断为精神分裂症的主题。 公共卫生相关性：对于大多数诊断为精神分裂症的患者，尚未确定直接的遗传原因。关于精神分裂症的一个重要理论表明，在某些大脑区域，如“前额叶皮层”，许多基因在正常发育过程中没有像健康受试者那样正常开启。然而，到目前为止，研究这些现象的分子技术一直缺乏。这项挑战性的赠款提案是“准备好了”，将创造新的就业机会，并保留现有的。它还基于我们实验室最近开发的极具创新性的技术。我们第一次能够从死后获得的人脑神经细胞中选择性地分离出染色体材料和染色质，并在全基因组水平上研究“表观遗传标记”（基本上是调节基因表达和功能而不改变遗传密码的化学修饰）。在这项提案中，我们计划检查正常前额叶皮层发育过程中的表观遗传基因激活模式，并在精神分裂症和突变小鼠中寻找潜在的改变，这些小鼠旨在在分子水平上模拟这种疾病。这项研究应该澄清表观遗传标记是否在人类神经元染色质中受到动态调节，以及精神分裂症等主要精神疾病是否与神经元表观基因组的变化有关。