Retrotransposons in Schizophrenia

精神分裂症中的反转录转座子

• 批准号: 9127614
• 负责人: Wade H Berrettini
• 金额: $ 67.73万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-21 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9127614
• 关键词: Adolescence; Affect; Alleles; Anterior; Antibodies; Autopsy; Biological Assay; Brain; Brain Diseases; Brain region; CRISPR/Cas technology; Cardiovascular Diseases; Cell Line; Cell Nucleus; Cell Separation; Cell membrane; Centrifugation; Chromatin; Chronic; Collaborations; Copy Number Polymorphism; DNA; DNA Sequence; Data; Daughter; Development; Disease; Doctor of Philosophy; Electroencephalography; Electrophysiology (science); Elements; Fluorescence; Gene Expression; Gene Frequency; Genes; Genetic; Genetic Transcription; Genome; Genomic DNA; Genomic Segment; Grant; Heritability; High-Throughput Nucleotide Sequencing; Human Genome; Inherited; Interneurons; Jumping Genes; L1 Elements; Label; Letters; Life; Life Expectancy; Membrane Proteins; Morbidity - disease rate; Mosaicism; Neurodevelopmental Disorder; Neurons; Nuclear; Odds Ratio; Ontology; Organism; Parvalbumins; Paste substance; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Persons; Population; Prefrontal Cortex; Procedures; Proteins; Psychotic Disorders; Public Health; RNA; RNA Polymerase II; RNA-Directed DNA Polymerase; Regulation; Rest; Retrotransposon; Risk; Sample Size; Sampling; Schizophrenia; Scientist; Site; Staining method; Stains; Suicide; Synapses; Technology; Testing; Tissues; Transcriptional Activation; Translations; Variant; base; digital; disability; emerging adult; endonuclease; gene function; genome wide association study; hippocampal pyramidal neuron; kindred; mortality; neuroimaging; promoter; public health relevance; reference genome; transposon/insertion element; young adult

 DESCRIPTION (provided by applicant): Schizophrenia (SZ) is a common, chronic group of psychotic brain disorders, affecting 1% of the US population, creating a significant public health problem because of the associated disability, morbidity and mortality. The pathogenesis of SZ is poorly understood, but it is thought to be a neurodevelopmental disorder. Neuroimaging evidence has accumulated to indicate that the dorsolateral prefrontal cortex (DLPFC) functions abnormally in SZ, both when activated and at rest. Further, EEG evidence has identified abnormalities of γ oscillations in SZ, suggesting that parvalbumin positive GABAergic interneuron regulation of cortical pyramidal neurons is dysfunctional. In the past 5 years, data have accumulated to prove that activation of LINE1 (L1) retrotransposons (RTPs) may occur wherever chromatin assumes a relaxed state to permit transcriptional activation. The abnormal DLPFC activation and γ oscillations in SZ may drive chronic aberrant transcriptional activation, thereby providing opportunities for L1s to retrotranspose in the developing CNS. These somatic de novo L1s may interfere with normal neuronal activity because they have inserted into a gene needed by that neuron for normal function. If one or more functional de novo L1s occur early in CNS development, all the daughter neurons that derive from that neuronal precursor will also carry the L1, perhaps leading to a dysfunctional population of neurons destined to increase risk for SZ. This project builds on preliminary results indicating that intragenic de novo L1s are found significantly more often in SZ DLPFC neuronal DNA in relevant gene ontologies (eg, synapse part and plasma membrane part; p = ~10-4), compared to control tissue. This proposal will employ DLPFC and anterior cingulate tissue (obtained at autopsy) from 100 SZ patients and 100 matched controls. Using differential centrifugation and fluorescence assisted cell sorting (FACS), followed by PCR to enrich the DNA for L1 sequences, DNA amplicons will be sequenced and aligned to the reference genome to detect intragenic de novo (not in the reference genome) L1s in brain-expressed genes. Allele frequencies of the most promising intragenic de novo L1s will be estimated by droplet digital PCR. Selected intragenic de novo L1s will be re-created in neuronal cell lines via CRISPR/Cas9 technology. The functional effect of the de novo L1 on transcription and translation of the gene will be determined. In this manner, it is expected that intragenic de novo L1s, which increase risk for SZ, will be discovered.

 描述（由适用提供）：精神分裂症（SZ）是一种常见的，慢性的精神病脑疾病，影响了1％的美国人群，由于相关的残疾，发病率和死亡率，造成了重大的公共卫生问题。 SZ的发病机理知之甚少，但被认为是一种神经发育障碍。神经影像学证据已积累，以表明在激活和静止时，SZ背侧前额叶皮层（DLPFC）的功能绝对。此外，脑电图证据已经鉴定出SZ中γ振荡的异常，表明白蛋白阳性GABA能性质中神经元的阳性对皮质金字塔神经元的调节功能失调。在过去的5年中，数据积累以证明Line1（L1）逆转录座子（RTP）的激活可能发生在染色质假定宽松状态以允许转录激活的情况下发生。 SZ中的异常DLPFC激活和γ振荡可能会驱动慢性异常转录激活，从而为L1s提供了在发育中的CNS中逆转转移的机会。这些从头开始的L1可能会干扰正常的神经元活性，因为它们已将神经元所需的基因插入正常功能。如果从中枢神经系统开发中出现一个或多个功能性的从头出现，那么从该神经元前体衍生的所有女儿神经元也将携带L1，也许会导致神经元的功能失调，这些神经元注定会增加SZ风险。该项目基于初步结果，表明与对照组织相比，在相关基因本体学（例如，突触部位和质膜部分； P = 〜10-4）中，SZ DLPFC神经元DNA中发现了基因内的L1s。该提案将采用DLPFC和100例SZ患者和100个匹配对照组的前扣带回组织（尸检）。使用差分离心和荧光辅助细胞分选（FACS），然后使用PCR富集L1序列的DNA，将对DNA扩增子进行测序并与参考基因组进行排列，以检测从头表达基因的基因组（不在参考基因组中）L1。最有希望的基因内L1的等位基因频率将由液滴数字PCR估算。选定的基因内L1S将通过CRISPR/CAS9技术在神经元细胞系中重新创建。将确定从头L1对基因转录和翻译的功能效应。以这种方式，将发现会发现SZ风险的从头开始L1。