Identifying critical functions of DISC1 disrupted in major mental illness

确定 DISC1 在重大精神疾病中受损的关键功能

• 批准号: 8829379
• 负责人: PRIYA SRIKANTH
• 金额: $ 3.5万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8829379
• 关键词: Adult; Affect; Biological Assay; Biology; Bipolar Disorder; Cell Proliferation; Cells; Characteristics; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Custom; Data; Development; Disease; Embryo; Equilibrium; Exons; Family; Gene Expression; Genetic; Genetic Engineering; Genomics; Human; Immigration; In Vitro; Interruption; Left; Length; Luciferases; Major Mental Illness; Measures; Mental disorders; Modeling; Molecular; Molecular Profiling; Morphology; Mutation; Neurites; Neuronal Differentiation; Neurons; Nonsense-Mediated Decay; Patients; Pattern; Play; Population; Predisposition; Process; Protein Isoforms; Publishing; RNA Splicing; Rattus; Reporter; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Schizophrenia; Signal Transduction; Site; Synapses; System; Transcript; Transcription Coactivator; Universities; Variant; Virus; Work; Xenograft procedure; base; gain of function; human disease; in vivo; induced pluripotent stem cell; loss of function; migration; mutant; nano-string; nerve stem cell; neural plate; nuclease; public health relevance; risk variant; stem

 DESCRIPTION (provided by applicant): Schizophrenia is a debilitating psychiatric disorder that affects ~1% of the world's population. Genetic and clinical association studies have identified disrupted-in-schizophrenia 1 (DISC1) as a strong candidate risk gene for schizophrenia and other major mental illnesses. DISC1 was initially associated with mental illness upon the discovery that its coding sequence is interrupted by a balanced chr(1;11) translocation in a Scottish family, in which the translocation cosegregates with schizophrenia, bipolar disorder and major depression1-3. DISC1 modulates many neuronal processes, including proliferation, Wnt signaling, synaptic maturation, neurite outgrowth, and neuronal migration4. Adding to the complexity of DISC1 biology, over 50 transcribed DISC1 splice variants have been identified in the developing and adult human brain5. The relevance of a DISC1 loss-of-function versus gain-of-function model to the human disease state is still unknown. As the expression patterns of DISC1 variants differ between humans and rodents, exploration of disease-relevant DISC1 expression and function is best accomplished in human cells. I aim to characterize the the expression profile of DISC1 during neuronal differentiation of human induced pluripotent stem (iPS) cells. I will further explore the functional consequences of interruption at the DISC1 locus in cells from t(1;11) patients and cells with total or disease-relevant DISC1-interruption resulting from transcription activator-like effector nucleases (TALENs) or the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system. DISC1 expression over neuronal differentiation will be assayed using a custom Nanostring probe set. Disease- relevant and total DISC1-disrupted cells will be differentiated to neural progenitor cells (NPCs) and cortical neurons and examined for altered cell fate, Wnt signaling, proliferation, morphology, and migration. My hypothesis is that interruption of DISC1 at the site of the Scottish translocation will result in nonsense- mediated decay of longer transcripts, leaving shorter transcripts intact, which in turn will disrupt a subset of DISC1-dependent processes. Based on published work and my preliminary data, I anticipate that disease-relevant DISC1 interruption will impair Wnt signaling and neuronal migration, but will not grossly alter gene expression or neurite length. In contrast, I expect that total DISC1 loss wil reduce neurite outgrowth in addition to altering Wnt signaling and neuronal migration. This study will reveal functions of DISC1 in human neural cells and highlight the functions of DISC1 disrupted by a disease-relevant mutation. These data will indicate those functions of DISC1 likely to be perturbed in patients with the chr(1;11) translocation and which, when disrupted, contribute to the development of mental illness.

 描述（由申请人提供）：精神分裂症是一种使人衰弱的精神疾病，影响着世界约 1% 的人口。遗传和临床关联研究已确定精神分裂症中断 1 (DISC1) 是精神分裂症和其他主要精神疾病的一个强有力的候选风险基因。 DISC1 最初与精神疾病相关，是因为在一个苏格兰家庭中发现其编码序列被平衡 chr(1;11) 易位打断，其中该易位与精神分裂症、双相情感障碍和重度抑郁症共分离1-3。 DISC1 调节许多神经元过程，包括增殖、Wnt 信号传导、突触成熟、神经突生长和神经元迁移4。进一步增加了 DISC1 生物学的复杂性，在发育中和成人大脑中已发现了 50 多种转录的 DISC1 剪接变体5。 DISC1 功能丧失与功能获得模型与人类疾病状态的相关性仍不清楚。由于人类和啮齿动物之间 DISC1 变体的表达模式不同，因此最好在人类细胞中探索与疾病相关的 DISC1 表达和功能。我的目的是表征 DISC1 在神经元分化过程中的表达谱 人类诱导多能干（iPS）细胞。我将进一步探讨 t(1;11) 患者细胞中 DISC1 位点中断的功能后果，以及由于转录激活因子样效应核酸酶 (TALEN) 或成簇规则间隔短回文重复序列 (CRISPR)-Cas 系统导致的完全或疾病相关 DISC1 中断的细胞。将使用定制的 Nanostring 探针组分析神经元分化中的 DISC1 表达。疾病相关细胞和 DISC1 破坏的总细胞将分化为神经祖细胞 (NPC) 和皮质神经元，并检查细胞命运、Wnt 信号传导、增殖、形态和迁移的改变。我的假设是，在苏格兰易位位点中断 DISC1 将导致较长转录本的无义介导的衰变，而较短的转录本完好无损，这反过来又会破坏 DISC1 依赖性过程的子集。根据已发表的工作和我的初步数据，我预计与疾病相关的 DISC1 中断将损害 Wnt 信号传导和神经元迁移，但不会严重改变基因表达或神经突长度。相比之下，我预计 DISC1 的全部缺失除了会改变 Wnt 信号传导和神经元迁移之外，还会减少神经突的生长。这项研究将揭示 DISC1 在人类神经细胞中的功能，并强调 DISC1 被疾病相关突变破坏的功能。这些数据将表明 DISC1 的功能可能在 chr(1;11) 易位患者中受到干扰，并且当受到干扰时，会导致精神疾病的发展。