Nemo-Like Kinase and the Pathogenesis of Spinal and Bulbar Muscular Atrophy

Nemo 样激酶与脊髓和延髓肌萎缩症的发病机制

基本信息

批准号：
8452770
负责人：
Tiffany Werlyne Todd
金额：
$ 4.22万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-30 至 2014-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8452770
关键词：
Address Affect Alleles Androgen Antagonists Androgen Receptor Androgens Atrophic Behavioral Assay Biochemical Biochemistry Biological Assay Biological Models CAG repeat Cell Culture System Cell Culture Techniques Cell Death Cell Line Cell model Cells Clinical Trials Confocal Microscopy Data Deglutition Disorders Development Disease Disease model Drosophila genus Drug Delivery Systems Dysarthria Face Female Fertility Future Genetic Genetic Techniques Genetic Transcription Glutamine Goals Gynecomastia Hela Cells Hormones Immunofluorescence Immunologic Kennedy Syndrome Lead Length Light Limb structure Link Luciferases Manuscripts Mediating Modeling Molecular Molecular Biology Molecular Genetics Motor Motor Neurons Muscle fasciculation Mutation Nature Nerve Degeneration Neurodegenerative Disorders Neuromuscular Junction Neurons Pathogenesis Pathology Patients Phenotype Phosphorylation Site Phosphotransferases Photoreceptors Protein-Serine-Threonine Kinases Proteins Quality of life Receptor Gene Role Spinal Spinal Cord Staining method Stains Symptoms System Techniques Testing Therapeutic Tissue Model Tissues Toxic effect Trinucleotide Repeats Type 1 Spinocerebellar Ataxia Western Blotting base cytotoxicity fly gain of function in vivo insight loss of function men middle age mutant nemo-like kinase neurodegenerative phenotype neurotoxicity novel novel therapeutics overexpression polyglutamine protein aggregate protein aggregation protein expression protein protein interaction spinal and bulbar muscular atrophy therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Spinal and Bulbar Muscular Atrophy (SBMA, Kennedy's Disease) is an X-linked progressive neurodegenerative disease affecting the proximal spinal and bulbar motoneurons. Patients present in midlife with weakness of the limbs and facial fasciculation that often progresses to dysarthria and dysphagia, complications of which can lead to fatality. SBMA patients also commonly suffer from mild androgen insensitivity, presenting with symptoms including gynecomastia, testicular atrophy, and decreased fertility. SBMA is caused by an expansion of a polymorphic CAG trinucleotide repeat in the Androgen Receptor (AR) gene, classifying the disease as one of the nine polyglutamine (polyQ) disorders. PolyQ expansion renders the host protein toxic, resulting in the formation of mutant protein aggregates and cell death. The commonalities in the nature of the mutation and the presentation of the different polyQ disorders suggest the presence of a common pathogenic mechanism. Nonetheless, this mechanism has remained elusive and to date there are no cures or even effective therapeutics for many of these diseases. Studying the pathogenesis of one polyQ disorder, namely SBMA, should hopefully shed some light on the pathogenic mechanisms underlying all nine diseases. Furthermore, the only SBMA-specific therapeutics to have undergone clinical trials are aimed at reducing the androgen insensitivity would be beneficial in maintaining patient quality-of-life. This proposal therefore investigates the role of the serine/threonine protein kinase Nemo-Like Kinase (NLK) in the pathogenesis of SBMA with the hope that NLK will prove a viable drug target in the development of future therapeutics. Specifically, this application addresses the hypothesis that NLK promotes the SBMA disease condition via interacting with and modulating the polyQ-expanded mutant AR. To test this idea, the effect of NLK co- expression on an SBMA cellular model will be investigated using protein expression, immunofluorescence, and cytotoxicity techniques. Both established and novel Drosophila models of SBMA will be employed to determine if there is a genetic interaction between NLK and the mutant AR using both photoreceptor neurons and the motor neurons/neuromuscular junction as model tissues. Finally, the molecular mechanism underlying the interaction between NLK and AR will be investigated using transcription assays, molecular biology and genetic techniques in cellular and Drosophila models. Through this integrated approach of genetics, molecular biology, and in vivo disease models, this proposal seeks to define the role of NLK in promoting the SBMA disease condition, paving the way for future mammalian studies and ultimately allowing for the development of novel, non-anti-androgen therapeutics to treat this devastating disorder. PUBLIC HEALTH RELEVANCE: The polyglutamine diseases represent a class of devastating neurodegenerative disorders that afflict millions of people worldwide. Despite the fact that these diseases arise from a similar mutation and share various symptomatic features, the underlying pathogenic mechanisms remain poorly understood and effective therapeutics are therefore unavailable for many of these conditions1. This proposal explores the role of Nemo- Like Kinase (NLK) in the pathogenesis of one polyglutamine disease, Spinal and Bulbar Muscular Atrophy, with the aim that NLK may prove to be a target protein in the development of novel therapeutics.

描述（由申请人提供）：脊髓和延髓肌萎缩症（SBMA，肯尼迪病）是一种影响近端脊髓和延髓运动神经元的 X 连锁进行性神经退行性疾病。患者在中年时出现四肢无力和面部肌束颤动，常常进展为构音障碍和吞咽困难，其并发症可能导致死亡。 SBMA 患者通常还患有轻度雄激素不敏感，表现为男性乳房发育、睾丸萎缩和生育能力下降等症状。 SBMA 是由雄激素受体 (AR) 基因中多态性 CAG 三核苷酸重复序列的扩增引起的，将该疾病归类为九种多聚谷氨酰胺 (polyQ) 疾病之一。 PolyQ 扩增使宿主蛋白有毒，导致突变蛋白聚集体的形成和细胞死亡。突变性质和不同 PolyQ 疾病表现的共性表明存在共同的致病机制。尽管如此，这种机制仍然难以捉摸，迄今为止，许多这些疾病还没有治愈方法，甚至没有有效的治疗方法。研究一种 PolyQ 疾病（即 SBMA）的发病机制，有望为所有九种疾病的致病机制提供一些线索。此外，唯一经过临床试验的 SBMA 特异性疗法旨在降低雄激素不敏感性，这将有利于维持患者的生活质量。因此，本提案调查了丝氨酸/苏氨酸蛋白激酶 Nemo 样激酶 (NLK) 在 SBMA 发病机制中的作用，希望 NLK 将成为未来治疗开发中可行的药物靶点。具体来说，本申请提出了这样的假设：NLK 通过与 polyQ 扩增的突变体 AR 相互作用并对其进行调节来促进 SBMA 疾病。为了测试这个想法，将使用蛋白质表达、免疫荧光和细胞毒性技术研究 NLK 共表达对 SBMA 细胞模型的影响。已建立的和新型的 SBMA 果蝇模型将用于确定 NLK 和突变 AR 之间是否存在遗传相互作用，使用感光神经元和运动神经元/神经肌肉接头作为模型组织。最后，将在细胞和果蝇模型中使用转录测定、分子生物学和遗传技术来研究 NLK 和 AR 之间相互作用的分子机制。通过这种遗传学、分子生物学和体内疾病模型的综合方法，该提案旨在明确 NLK 在促进 SBMA 疾病中的作用，为未来的哺乳动物研究铺平道路，并最终允许开发新颖的、非抗雄激素疗法来治疗这种破坏性的疾病。公共卫生相关性：多聚谷氨酰胺疾病代表了一类毁灭性的神经退行性疾病，困扰着全世界数百万人。尽管这些疾病源于相似的突变并具有不同的症状特征，但其潜在的致病机制仍然知之甚少，因此对于其中许多疾病尚无有效的治疗方法1。该提案探讨了 Nemo 样激酶 (NLK) 在一种多聚谷氨酰胺疾病——脊髓和延髓肌萎缩症的发病机制中的作用，目的是 NLK 可能被证明是开发新疗法的靶蛋白。