Core B

核心B

基本信息

批准号：
9304366
负责人：
Jeffrey T. Leek
金额：
$ 13.82万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9304366
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 Imagery 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 Work base brain tissue cohesion 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

项目摘要

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的突变引起的转录本表达式变化与精神分裂症（SZ）相关的基因以及人类细胞和小鼠模型中的压力暴露，在青少年人类大脑中特异性调节的基因之间富集，并在精神病疾病，例如SZ。 AIM 1将检查IPS细胞中的转录本表达变化 - 由CRISPR/CAS9介导的KCTD13，DPYSL2，DPYSL2介导的诱变产生的人类神经元细胞通过进行RNA-seq分析，SDCCAG8和CKAP5。我们还将使用这些遗传细胞模型确定有或不用地塞米松治疗的转录本表达的变化作为模型体外压力暴露。 AIM 2将检查前额叶皮层（PFC）的转录本表达变化 PCM1敲除小鼠，16P11（DUP）小鼠模型，16p11（dup）/kctd13 +/-小鼠以及其他鼠标模型，例如 DPYSL2通过进行RNA-seq分析来条件敲除小鼠。我们还将检查是否观察到青少年的社会隔离会加剧变化。 AIM 3将比较基因组区域和成绩单在AIM 1和2中鉴定为由于基因突变和细胞中的压力暴露而差异表达的小鼠模型与大规模尸体后人脑组织RNA-Seq中鉴定出的转录本和基因收藏。使用独特的人类验尸RNA-seq数据集，我们将解决我们的发现是否可以是在与人类青少年大脑成熟和精神病疾病有关的两种情况下翻译。我们还将在前瞻性人的血清中交叉验证差异表达的应激相关分子受试者开发了由基因突变和社会隔离以及小鼠模型的青少年人类大脑和PFC区域。