Altered reverse transcriptase-dependent gene diversification mechanisms in Alzheimer's disease brains

阿尔茨海默病大脑中逆转录酶依赖性基因多样化机制的改变

基本信息

批准号：
10545795
负责人：
JEROLD CHUN
金额：
$ 10.06万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10545795
关键词：
Abbreviations Affect Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease therapeutic Alzheimer&apos s disease therapy Amyloid beta-Protein Precursor Animal Experiments Animal Model Autopsy Back Biological Models Biological Sciences Brain Brain region Brodmann&apos s area Cell model Complementary DNA Consensus Sequence Copy Number Polymorphism DNA DNA strand break Data Disease Down Syndrome FDA approved Female Fishes Fluorescence Fluorescent in Situ Hybridization Foundations Gene Proteins Genes Genetic Recombination Genetic Transcription Genome Genomic DNA Genomics Green Fluorescent Proteins HIV Human IACUC In Situ Hybridization Institutional Review Boards Knowledge Methodology Modeling Molecular Molecular Biology Molecular Neurobiology Molecular Target Mosaicism Nature Neurofibrillary Tangles Neuroglia Neurons Nuclear Nucleotides Pathogenicity Patients Pattern Peptide Nucleic Acids Ploidies Prevalence Publishing Quality Control RNA RNA-Directed DNA Polymerase Reporting Reverse Transcriptase Inhibitors Reverse Transcription Role Sampling Senile Plaques Sorting - Cell Movement Testing Therapeutic Toxic effect Validation Variant aged brain tissue cell type ds-DNA familial Alzheimer disease gene product genetic variant human embryonic stem cell induced pluripotent stem cell inhibitor insertion/deletion mutation integration site male nanobodies neurobiological mechanism new therapeutic target next generation sequencing novel sequencing platform sex single molecule real time sequencing therapeutic target

项目摘要

PROJECT SUMMARY/ABSTRACT We have identified somatic gene recombination (SGR) in neurons of the human brain, with particular relevance to sporadic Alzheimer’s disease (SAD) (Nature 563, 639-645 (2018)). This discovery represents a new and functionally significant aspect of genomic mosaicism (eLife;4:e05116 (2015)) that has genuine therapeutic potential through newly identified molecular targets. Indeed, we found that SGR, acting on the AD gene for Amyloid Precursor Protein (APP), produces thousands of distinct forms of APP, some of which are enriched in or unique to AD. The APP gene variations related to AD that were analyzed thus far include copy number variations (CNVs) and at least 11 single-nucleotide variations (SNVs) that were previously reported as pathogenic in familial AD, yet that arose somatically and mosaically in SAD; these variations were absent from non-diseased neurons. SGR utilizes reverse transcriptase (RT) activity on transcribed RNAs that, combined with DNA strand-breaks and APP gene transcription, produce double-stranded DNA that is retro-inserted back into the genome to form “genomic cDNAs” (gencDNAs). These published data contribute to the scientific foundation on which the current proposal will build, to test the hypothesis that altered SGR, involving brain- specific reverse transcriptases, functionally contributes to AD and affects multiple genes, providing novel targets for AD therapies. Postmortem IRB-approved and de-identified brain samples from validated AD donors of both sexes will be compared to non-diseased controls, while IACUC-approved animal experiments will model SGR and its AD-relevant endpoints. Three Aims will be pursued over 5 years. Aim 1 will define the molecular neurobiology of APP gencDNA diversity and identify new SGR genes enhanced in AD brains. Aim 2 will determine expression and function of SGR genes in AD brain and model systems. Aim 3 will identify genes responsible for RT SGR activity within normal and AD brains. This proposal will thus open new vistas into AD via novel SGR mechanisms and will identify new therapeutic targets for the treatment of AD.

项目摘要/摘要我们已经确定了人脑神经元中的体细胞基因重组（SGR），特别相关偶发的阿尔茨海默氏病（SAD）（自然563，639-645（2018））。该发现代表了一个新的基因组镶嵌的功能重要方面（Elife; 4：E05116（2015））具有真正的治疗通过新确定的分子靶标的潜力。确实，我们发现SGR在广告基因上的作用淀粉样蛋白前体蛋白（APP）产生数千种不同形式的应用程序，其中一些富集在或广告独有的。到目前为止对与AD相关的APP基因变异包括拷贝数以前报道为家族性广告中的致病性，但在SAD上出现了体形和镶嵌性。这些变化是从中出现的未切除的神经元。 SGR在转录的RNA上使用逆转录酶（RT）活性，该活性合并使用DNA链断裂和APP基因转录，产生双链DNA，恢复后插入进入基因组，形成“基因组cDNA”（GENCDNA）。这些已发布的数据有助于科学当前建议将建立的基础，以检验改变SGR的假设，涉及大脑特定的逆转录酶，在功能上有助于AD并影响多个基因，提供了新的靶标用于广告疗法。验尸IRB批准并取消识别的大脑样本性别将与未解决的对照进行比较，而IACUC批准的动物实验将建模SGR 及其与广告相关的终点。三个目标将在5年内实现。 AIM 1将定义分子 App GencDNA多样性的神经生物学，并确定AD大脑中增强的新SGR基因。 AIM 2意志确定SGR基因在AD大脑和模型系统中的表达和功能。 AIM 3将识别基因负责正常和广告大脑内的RT SGR活动。因此，该提案将向广告开放新的远景通过新型的SGR机制，将确定用于治疗AD的新治疗靶标。