Pathophysiology of DYT1 dystonia: Targeted Mouse Models

DYT1 肌张力障碍的病理生理学：靶向小鼠模型

基本信息

批准号：
10710411
负责人：
YUQING LI
金额：
$ 36.29万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-28 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10710411
关键词：
Acetylcholine Affect American Amino Acids Animal Model Antisense Oligonucleotides Artificial Intelligence Behavior Behavioral Binding Biochemical Brain Brain region Cells Characteristics Clustered Regularly Interspaced Short Palindromic Repeats Corpus striatum structure DRD2 gene DYT1 gene Development Disabled Persons Disease Dopamine Dopamine Receptor Dystonia Early Onset Dystonia Electrophysiology (science)Endoplasmic Reticulum Environment Exons Family Functional disorder Funding GAG Gene Genes Genetic Glutamates Goals Impairment Individual Interneurons Knock-in Knock-in Mouse Knockout Mice Knowledge Lead Limb structure Machine Learning Metabolism Modeling Molecular Chaperones Molecular Genetics Movement Movement Disorders Mus Muscle Contraction Neuronal Dysfunction Neurons Newborn Infant Nuclear Envelope Nucleotides Parkinson Disease Pathogenesis Pathway interactions Patients Penetrance Phenotype Play Posture Proteins Research Research Personnel Role Source Symptoms System TOR1A gene Techniques Testing TorsinA Tremor United States National Institutes of Health Wheelchairs Work autosome brain cell cell type cholinergic cholinergic neuron conditional knockout dopaminergic neuron early onset effective therapy gain of function in vivo interdisciplinary approach motor deficit mouse model network models neurochemistry novel protein folding protein function small hairpin RNA targeted treatment therapeutically effective

项目摘要

Dystonia is a movement disorder characterized by sustained or intermittent muscle contractions causing abnormal, often repetitive, movements or postures. DYT1 early-onset generalized dystonia is the most common type among the genetic dystonias. Most of the individuals affected by DYT1 dystonia share a trinucleotide deletion (ΔGAG) located in exon 5 of DYT1 or TOR1A gene, leading to a loss of a glutamate amino acid residue for torsinA (torsinA∆E). The symptoms start from limbs and then become generalized. Affected individuals could be seriously disabled and need to use a wheelchair. Conditional knockout of torsinA in mice points to the involvement of multiple brain regions and cell types in the pathogenesis of DYT1 dystonia. These and other pathophysiological studies of DYT1 and other dystonias so far support a circuit or network model of dystonia pathogenesis. However, which brain region and neuronal types play a critical role in pathogenesis is unclear. Alterations of both the striatal dopaminergic and cholinergic systems appear to play a critical role in the pathophysiology of DYT1 dystonia. Dopaminergic modulation of striatal cholinergic interneurons (ChIs) is altered in multiple dystonia models that include DYT1 dystonia. Whether torsinA∆E in striatal ChIs has the cell-autonomous effect on striatal cholinergic dysfunction is not known. Preliminary studies of conditional knockin (KI) mouse models of DYT1 dystonia revealed dopaminergic and striatal medium spiny neurons (MSNs), but not ChIs, play a vital role in the pathogenesis of DYT1 dystonia. However, how torsinA∆E in MSNs and dopaminergic neurons lead to dystonia is unknown. These unknowns impede the progress in developing effective treatment for DYT1 patients, especially gene-based targeted therapy with CRISPR, antisense oligonucleotides, or small hairpin RNA. The broad, long-term objective of our research is 1) to determine the functional role of torsinA and the mechanism by which torsinA∆E leads to early-onset dystonia, 2) to develop novel and effective therapeutic treatment. The specific goal of this application is to generate and analyze five lines of conditional KI mice to understand the role of the striatal dopaminergic system and MSNs in the pathogenesis of DYT1 dystonia. We hypothesize that torsinA∆E in dopaminergic neurons and MSNs, but not ChIs, leads to abnormal firing of these neurons, decreased D1R and D2R in MSNs, altered striatal dopamine release, impaired corticostriatal LTD, altered direct and indirect pathways, and ultimately sustained muscle contractions and co-contractions that are characteristic of dystonia. Aim 1 will generate conditional MSN KI mice restricted to direct pathway, indirect pathway, or both and determine their phenotype. In Aim 2, we will introduce torsinA∆E specifically in dopaminergic neurons or ChIs and determine their dystonia-related phenotypes. The successful completion of the proposed research will significantly increase our understanding of the pathophysiology of DYT1 dystonia and aid the development of novel targeted treatments for dystonia patients.

肌张力障碍是一种运动障碍，其特征是持续或间歇性肌肉收缩引起异常，通常重复的运动或位置。 DYT1早期发表的广义肌张力障碍是最大的遗传性肌张力障碍的常见类型。受dyt1肌张力障碍影响的大多数人共享位于dyt1或tor1a基因外显子5中的三核苷酸缺失（ΔGAG），导致谷氨酸的损失 Torsina（TorsinaΔE）的氨基酸住所。症状从四肢开始，然后泛化。受影响的人可能会严重残疾，需要使用轮椅。有条件的托尔西娜在小鼠中，表明多个大脑区域和细胞类型参与DYT1肌张力障碍的发病机理。到目前为止，这些DYT1和其他肌张力的病理生理研究支持电路或网络肌张力障碍发病机理的模型。但是，哪种大脑区域和神经元类型在发病机理尚不清楚。纹状体多巴胺能和胆碱能系统的改变似乎都在发挥在DYT1肌张力障碍的病理生理学中的关键作用。纹状体胆碱能的多巴胺能调节在包括dyt1肌张力障碍的多种肌张力障碍模型中，中间神经元（CHIS）发生了变化。是否torsinaΔeIN 纹状体CHI对纹状体胆碱能功能障碍具有细胞自主作用，尚不清楚。初步研究 Dyt1肌张力障碍的条件性敲击（Ki）小鼠模型揭示了多巴胺能和纹状体培养基神经元（MSN）而不是CHI，在Dyt1 dystonia的发病机理中起着至关重要的作用。但是，torsinaΔe 在MSN中，多巴胺能神经元导致肌张力障碍是未知的。这些未知数阻碍了进度为DYT1患者开发有效治疗，尤其是基于基因的基于基因的针对性治疗CRISPR，反义寡核苷酸或小发夹RNA。我们研究的广泛长期目标是1）确定Torsina的功能作用以及Torsina∆E导致早期发作肌张力障碍的机制，2）开发新颖有效的治疗治疗。此应用程序的具体目标是生成和分析有条件的Ki小鼠的五行，以了解纹状体多巴胺能系统和MSN的作用在Dyt1肌张力障碍的发病机理中。我们假设多巴胺能神经元和MSN中的TorsinaΔE，但是不是CHI，导致这些神经元的异常发射，降低了MSN的D1R和D2R，改变了纹状体多巴胺释放，皮质纹状体有限公司受损，改变了直接和间接途径，并最终持续肌肉收缩和共同收缩是肌张力障碍的特征。 AIM 1将产生有条件的 MSN Ki小鼠仅限于直接途径，间接途径或两者兼而有之并确定其表型。在AIM 2中，我们将在多巴胺能神经元或CHI中专门介绍TorsinaΔE，并确定其肌张力障碍有关表型。拟议研究的成功完成将大大提高我们的理解 Dyt1肌张力障碍的病理生理学，并有助于开发新的靶向肌张力障碍治疗方法患者。