Retrotransposons in Schizophrenia

精神分裂症中的反转录转座子

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：精神分裂症 (SZ) 是一种常见的慢性精神性脑部疾病，影响 1% 的美国人口，由于相关的残疾、发病率和死亡率，造成了重大的公共健康问题。人们对它知之甚少，但它被认为是一种神经发育障碍，神经影像学证据表明，背外侧前额叶皮层（DLPFC）在激活时和处于激活状态时功能异常。此外，脑电图证据发现 SZ 中 γ 振荡异常，表明皮质锥体神经元的小白蛋白阳性 GABA 能中间神经元调节功能失调。在过去 5 年中，积累的数据证明 LINE1 (L1) 逆转录转座子 (RTP) 的激活。）可能发生在染色质处于松弛状态以允许转录激活的地方。异常的 DLPFC 激活和 SZ 中的 γ 振荡可能会导致慢性。异常转录激活，为 L1 在由此发育的 CNS 中逆转录提供机会，这些体细胞从头 L1 可能会干扰正常神经活动，因为它们已插入该神经元正常功能所需的基因。发生在中枢神经系统发育早期，所有源自该神经前体的子神经元也将携带 L1，可能会导致神经元群体功能失调，从而增加 SZ 的风险。该项目建立在初步结果的基础上，表明：与对照组织相比，在 SZ DLPFC 神经元 DNA 的相关基因本体（例如，突触部分和质膜部分；p = ~10-4）中，基因内从头 L1 的发现频率明显更高。该提案将采用 DLPFC 和前扣带回组织。（尸检时获得）来自 100 名 SZ 患者和 100 名匹配对照，使用差速离心和荧光辅助细胞分选 (FACS)，然后通过 PCR 富集 DNA。 L1 序列、DNA 扩增子将被测序并与参考基因组进行比对，以检测脑表达基因中的基因内从头 (不在参考基因组中) L1。最有希望的基因内从头 L1 的等位基因频率将通过液滴数字 PCR 进行估计。将通过 CRISPR/Cas9 技术在神经元细胞系中重新创建选定的基因内 de novo L1，并确定 de novo L1 对基因转录和翻译的功能影响。通过这种方式，预计会发现增加 SZ 风险的基因内从头 L1。