The role of ADAR2-associated RNA editing in pathogenesis of ALS

ADAR2 相关 RNA 编辑在 ALS 发病机制中的作用

基本信息

批准号：
10084222
负责人：
STELLA DRACHEVA
金额：
--
依托单位：
JAMES J PETERS VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-10-01 至 2021-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10084222
关键词：
ALS patients AMPA Receptors Acids Age Amyotrophic Lateral Sclerosis Animals Astrocytes Autopsy Biological Assay Brain region C9ORF72 CRISPR/Cas technology Calcium Cells Coculture Techniques DRADA2b protein Data Disease Disease Pathway Disease Progression Down-Regulation Enzymes Event Family history of Fluorescence-Activated Cell Sorting Gene Expression Genes Glutamates Goals Healthcare Systems Heterogeneity Human Immunohistochemistry Inflammatory Inflammatory Response Inherited Kainic Acid Receptors Knowledge Laboratories Lasers Massive Parallel Sequencing Measures Mediating Messenger RNA Methods Microglia Molecular Motor Cortex Motor Neurons Muscle Weakness Mutation Nature Neurodegenerative Disorders Neurons Nuclear Antigens Occipital lobe Oligodendroglia Paralysed Pathogenesis Pathologic Patients Population Potassium Process Publishing Quality of life RNA Editing Reporting Research Risk Factors Role Serotonin Receptor 5-HT2C Signal Transduction Specimen Spinal Spinal Cord Spinal cord injury System Testing Time Tissues Transcript Transcription Alteration Variant Veterans base cell type cytokine differential expression improved induced pluripotent stem cell innovation male military service neuroinflammation neuronal excitability novel nucleocytoplasmic transport receptor restriction enzyme sex sporadic amyotrophic lateral sclerosis stem superoxide dismutase 1 transcriptome transcriptome sequencing transmission process

项目摘要

Amyotrophic Lateral Sclerosis (ALS) is a devastating and fatal neurodegenerative disease that is characterized by progressive muscle weakness and eventual paralysis. The known risk factors for ALS are increasing age and male sex; recent data also implicate military service, thus putting an additional burden on the VA healthcare system. Approximately 90% of all ALS cases are classified as sporadic ALS (sALS) with no family history of the disease. The remaining cases are inherited in a dominant fashion, and mutations in ~30 genes (most commonly in C9orf72, SOD1, TARDBP, FUS) are currently known to cause ALS. Hyperexcitability that stems from pathophysiological disturbance of glutamatergic transmission has been reported in all types of ALS cases, and may be a common disease pathway that predisposes motoneurons to degeneration. It has been suggested that the glutamatergic (Glu) deficit in ALS may result from altered function of the AMPA Glu receptors due to inefficient RNA editing of the GluA2 AMPA subunit. The GluA2 editing is catalyzed by the editing enzyme ADAR2. Our data, as well as data published by others, showed the downregulation of ADAR2 and inefficient editing of the GluA2 in the motor cortex of patients with C9orf72-associated ALS (C9 ALS) as well as in spinal motoneurons of patients with sALS. We also reported the downregulation of ADAR2 and the consequent decrease of editing in several of its targets in animals subjected to spinal cord injury (SCI). Moreover, in SCI these events are triggered by inflammatory response and result in changes in gene expression that are likely to contribute to post-SCI motoneuron hyperexcitability. Based on these data and the fact that, as in SCI, neuroinflammation is present in ALS, we hypothesize that: (1) ADAR2 is downregulated in both C9 ALS and sALS, leading to alterations in RNA editing in its targets. In particular, in addition to GluA2, ADAR2 editing of other molecules which are important for neuronal excitability [such as, GluA3-4, kainate receptor subunits GluK1-2, serotonin 2C receptor (5-HT2CR), as well as potassium (Kv1.1) and calcium (Cav1.3) channels] is also altered in C9 ALS and sALS; (2) The downregulation of ADAR2 in ALS results from several different pathological features associated with the disease. Whereas the decrease of ADAR2 in sALS is triggered by neuroinflammation, the availability of the functional ADAR2 in C9 ALS is also influenced by its cytoplasmic retention, which is caused by a disruption of nucleocytoplasmic transport. We will test these hypotheses by characterizing ALS-associated alterations (1) in the ADAR2 expression and localization and (2) in editing of the transcripts that are targeted by ADAR2. These studies will be performed in laser microdissected neurons obtained from autopsy tissues of patients with C9 and sporadic ALS (Aim1) as well as in motoneurons differentiated from human induced pluripotent stem cells (hiPSC) derived from ALS patients (Aim2). In addition, previously published studies of ALS-associated transcriptional alterations employed whole (cellular heterogeneous) tissue specimens. However, because of the heterogeneity of cell types in the CNS, accurate assessment of the ALS transcriptome cannot be inferred from the data obtained from these specimens that combine signals from all cell types. Therefore, we will study cell- type-specific transcriptomes in C9 ALS and sALS (Aim3). These studies are enabled by the novel methods recently developed in our lab, and should enhance the likelihood of elucidating disease relevant variations, including those related to ADAR2 and editing. Overall, this is an innovative proposal that builds on the interdisciplinary nature of our collaborative team to investigate RNA editing changes that characterize ALS and to define unique changes occurring only in C9 or sporadic ALS. Our goal is to advance the current knowledge and to discover new treatments.

肌萎缩性脊髓侧索硬化症 (ALS) 是一种毁灭性且致命的神经退行性疾病，其特征是进行性肌肉无力并最终瘫痪。 ALS 的已知危险因素是年龄和男性性别的增加；最近的数据还涉及兵役，从而给人们带来了额外的负担退伍军人管理局医疗保健系统。大约 90% 的 ALS 病例被归类为散发性 ALS (sALS)，无症状该疾病的家族史。其余病例以显性方式遗传，约 30 例发生突变目前已知基因（最常见的是 C9orf72、SOD1、TARDBP、FUS）会导致 ALS。源于谷氨酸能传递病理生理学紊乱的过度兴奋在所有类型的 ALS 病例中都有报道，并且可能是易感的常见疾病途径运动神经元退化。有人提出，ALS 中的谷氨酸 (Glu) 缺乏可能会导致由于 GluA2 AMPA 亚基的 RNA 编辑效率低下，AMPA Glu 受体的功能发生改变。 GluA2 编辑由编辑酶 ADAR2 催化。我们的数据以及其他人发布的数据，显示 ADAR2 下调和 GluA2 在运动皮层患者中的低效编辑 C9orf72 相关的 ALS (C9 ALS) 以及 sALS 患者的脊髓运动神经元。我们还报道了 ADAR2 的下调以及随之而来的动物体内几个靶标的编辑减少遭受脊髓损伤（SCI）。此外，在 SCI 中，这些事件是由炎症反应引发的并导致基因表达的变化，可能导致 SCI 后运动神经元过度兴奋。根据这些数据以及 ALS 中存在神经炎症（与 SCI 一样）的事实，我们假设 (1) ADAR2 在 C9 ALS 和 sALS 中均下调，导致其靶点的 RNA 编辑发生变化。特别是，除了 GluA2 之外，ADAR2 还可以编辑对神经元重要的其他分子。兴奋性 [例如，GluA3-4、红藻氨酸受体亚基 GluK1-2、5-羟色胺 2C 受体 (5-HT2CR)，以及钾 (Kv1.1) 和钙 (Cav1.3) 通道] 在 C9 ALS 和 sALS 中也发生改变； (2) 下调 ADAR2 在 ALS 中的表达是由与该疾病相关的几种不同病理特征引起的。鉴于 sALS 中 ADAR2 的减少是由神经炎症触发的，C9 中功能性 ADAR2 的可用性 ALS 还受到其细胞质滞留的影响，这是由核细胞质破坏引起的运输。我们将通过描述 ADAR2 中与 ALS 相关的改变 (1) 来测试这些假设表达和定位以及 (2) 编辑 ADAR2 靶向的转录本。这些研究将在从 C9 和散发性患者尸检组织中获得的激光显微切割神经元中进行 ALS (Aim1) 以及从人类诱导多能干细胞 (hiPSC) 分化而来的运动神经元源自 ALS 患者（目标2）。此外，之前发表的 ALS 相关转录研究改变采用整个（细胞异质）组织样本。然而，由于 CNS 细胞类型的异质性，无法推断 ALS 转录组的准确评估从这些样本中获得的数据结合了所有细胞类型的信号。因此，我们将研究细胞 C9 ALS 和 sALS 中的类型特异性转录组 (Aim3)。这些研究是通过新颖的方法实现的我们实验室最近开发的，应该增强阐明疾病相关变异的可能性，包括与 ADAR2 和编辑相关的内容。总的来说，这是一项创新提案，建立在我们合作的跨学科性质之上。团队研究了 ALS 特征的 RNA 编辑变化，并定义了仅发生在 C9 或散发性 ALS。我们的目标是推进现有知识并发现新的治疗方法。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Divergent single cell transcriptome and epigenome alterations in ALS and FTD patients with C9orf72 mutation.

DOI：
10.1038/s41467-023-41033-y
发表时间：
2023-09-15
期刊：
NATURE COMMUNICATIONS
影响因子：
16.6
作者：
Li, Junhao;Jaiswal, Manoj K.;Chien, Jo-Fan;Kozlenkov, Alexey;Jung, Jinyoung;Zhou, Ping;Gardashli, Mahammad;Pregent, Luc J.;Engelberg-Cook, Erica;Dickson, Dennis W.;Belzil, Veronique V.;Mukamel, Eran A.;Dracheva, Stella
通讯作者：
Dracheva, Stella