Core B

核心B

• 批准号: 9978143
• 负责人: Jeffrey T. Leek
• 金额: $ 12.62万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-12 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978143
• 关键词: Address; Adolescence; Adolescent; Animal Model; Autopsy; Base Pairing; Brain; CRISPR/Cas technology; Cell model; Cells; Collaborations; Collection; Computer software; DNA Sequence Alteration; Data; Data Analyses; Data Set; Development; Dexamethasone; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Genome Scan; Genomic Segment; Goals; Human; In Vitro; Induced Mutation; Knockout Mice; Mediating; Mental disorders; Microtubules; Modeling; Mus; Mutagenesis; Neurons; Pathway Analysis; Prefrontal Cortex; Protein Isoforms; Quality Control; Resolution; Resources; Schizophrenia; Serum; Social isolation; Standardization; Statistical Models; Stress; Transcript; Translating; Update; Visualization; Work; analysis pipeline; base; brain tissue; cohesion; conditional knockout; critical period; design; differential expression; gene environment interaction; genetic variant; human subject; induced pluripotent stem cell; innovation; mouse model; novel; prospective; reference genome; transcriptome; transcriptome sequencing

Our main goal is to identify the transcript expression changes induced by mutations in microtubule-related genes associated with schizophrenia (SZ) and stress exposure in human cell and mouse models, which are enriched among genes specifically regulated in adolescent human brains and differentially expressed in psychiatric disease conditions, such as SZ. Aim 1 will examine the transcript expression changes in iPS cells- derived human neuronal cells resulting from CRISPR/Cas9 mediated mutagenesis of KCTD13, DPYSL2, SDCCAG8 and CKAP5 by conducting a RNA-seq analysis. We will also use these genetic cell models to determine changes in transcript expression with and without treatment with dexamethasone as a model of stress exposure in vitro. Aim 2 will examine the transcript expression changes in the prefrontal cortex (PFC) of Pcm1 knockout mice, 16p11(dup) mice model, 16p11(dup)/Kctd13+/- mice, and other mouse models such as Dpysl2 conditional knockout mice by conducting a RNA-seq analysis. We will also examine if observed changes are exacerbated by adolescent social isolation. Aim 3 will compare genomic regions and transcripts identified in Aims 1 and 2 as differentially expressed due to genetic mutations and stress exposure in cell and mouse models to transcripts and genes identified in large-scale postmortem human brain tissue RNA-seq collections. Using unique human post-mortem RNA-seq datasets, we will address whether our findings can be translated in two contexts relevant to human adolescent brain maturation and psychiatric disease conditions. We will also cross-validate differentially expressed stress-associated molecules in serum of prospective human subjects developed SZ and mouse models resulting from genetic mutations and social isolation as well as adolescent human brains and PFC region of the mouse models.

我们的主要目标是确定微管相关基因突变引起的转录表达变化 人类细胞和小鼠模型中与精神分裂症(SZ)和应激暴露相关的基因，这些基因是 在青春期人脑中特异调控的基因中丰富，并在 精神疾病状况，如SZ。Aim 1将研究iPS细胞中转录表达的变化- CRISPR/Cas9诱变KCTD13、DPYSL2、 SDCCAG8和CKAP5通过进行RNA-seq分析。我们还将使用这些遗传细胞模型来 以地塞米松为模型，检测地塞米松治疗前后转录表达的变化 体外应激暴露。目标2将检测大鼠前额叶皮质(PFC)转录表达的变化 Pcm1基因敲除小鼠、16p11(DUP)小鼠模型、16p11(DUP)/KCTD13+/-小鼠以及其他小鼠模型，如 Dpysl2条件性基因敲除小鼠进行RNA-seq分析。我们还将检查是否观察到 青少年的社交孤立加剧了这种变化。目标3将比较基因组区域和转录本 在AIMS 1和AIMS 2中发现由于遗传突变和细胞和 小鼠模型到大规模人脑组织中发现的转录和基因的RNA-seq 收藏。使用独特的人类死后rna-seq数据集，我们将解决我们的发现是否可以 在两个与人类青春期大脑成熟和精神疾病状况相关的背景下翻译。 我们还将交叉验证未来人类血清中差异表达的应激相关分子。 受试者开发了基因突变和社会隔离导致的SZ和小鼠模型，以及 青春期人脑和小鼠PFC区模型。