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由微管相关基因突变引起的转录本表达的变化。 在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， 在青少年大脑中特异性调控的基因中富集，并在 精神疾病，如SZ。目的1将检测iPS细胞中转录本表达的变化- 来源于CRISPR/Cas9介导的KCTD 13，DPYSL 2， SDCCAG 8和CKAP 5的基因组序列。我们还将使用这些遗传细胞模型， 确定用和不用地塞米松治疗的转录物表达的变化，作为 体外应力暴露。目的2将检测在脑缺血再灌注损伤后， Pcm 1敲除小鼠、16 p11（dup）小鼠模型、16 p11（dup）/Kctd 13 +/-小鼠和其他小鼠模型，例如 Dpysl 2条件性敲除小鼠通过进行RNA-seq分析。我们还将检查是否观察到 青少年的社会孤立加剧了这些变化。目标3将比较基因组区域和转录本 在目的1和2中鉴定为由于细胞中的基因突变和应激暴露而差异表达， 在大规模死后人脑组织中鉴定的转录物和基因的小鼠模型RNA-seq 收藏.使用独特的人类死后RNA-seq数据集，我们将讨论我们的发现是否可以用于 在两个与人类青少年大脑成熟和精神疾病状况相关的背景下翻译。 我们还将交叉验证在未来的人血清中差异表达的应激相关分子， 受试者开发了SZ和小鼠模型，这些模型是由基因突变和社会隔离以及 青少年人脑和小鼠PFC区模型。