Utilizing microRNA in cerebrospinal fluid to evaluate the effects of tyrosine kinase inhibition via Nilotinib on gene regulation in individuals with Parkinson's disease or Alzheimer's disease

利用脑脊液中的 microRNA 评估尼洛替尼抑制酪氨酸激酶对帕金森病或阿尔茨海默病患者基因调控的影响

• 批准号: 10201534
• 负责人: Alan Fowler
• 金额: $ 2.91万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2021-08-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201534
• 关键词: 3,4-Dihydroxyphenylacetic Acid; Affect; Age-Years; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease therapy; Amyloid beta-Protein; Animal Model; Attenuated; Autophagocytosis; Biological; Biological Assay; Biological Markers; Blood - brain barrier anatomy; Brain; Cerebrospinal Fluid; Cerebrospinal Fluid Proteins; Clinical; Clinical Trials; Cognition; Computer software; Data; Dementia with Lewy Bodies; Development; Disease; Disease Progression; Disinhibition; Dopamine; Dose; Down-Regulation; Enrollment; Enzyme-Linked Immunosorbent Assay; Evaluation; FYN gene; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Homovanillic Acid; Human; Impaired cognition; Individual; Intervention; Liquid substance; Lysosomes; Measures; Metabolism; MicroRNAs; Motor; Nerve Degeneration; Neurodegenerative Disorders; Parkinson Disease; Parkinson' s Dementia; Pathology; Pathway Analysis; Pathway interactions; Patients; Phase; Pilot Projects; Plasma; Polymerase Chain Reaction; Population; Post-Transcriptional Regulation; Protein Tyrosine Kinase; Proteins; Proteolysis; Reproducibility; Reverse Transcription; Role; Surrogate Markers; TREM2 gene; Testing; Time; Tyrosine Kinase Inhibitor; Ubiquitination; United States; Untranslated RNA; Work; alpha synuclein; cohort; data mining; differential expression; discoidin receptor; experimental study; improved; individual response; inhibitor/antagonist; miRNA expression profiling; multicatalytic endopeptidase complex; neuroinflammation; neuropathology; neurotoxic; next generation sequencing; non-motor symptom; open label; pre-clinical; protein biomarkers; response; response biomarker; tau Proteins; tau-1; treatment comparison

Project summary/Abstract: Alzheimer’s disease (AD) and Parkinson’s disease (PD), are estimated to affect 40% and 1% of the United Stated population over 85 years of age, respectively, making them the two most common neurodegenerative diseases [1-6]. The identification of a reliable biomarker for response to interventions in AD and PD will greatly assist in the development of successful disease modifying therapies. Our strong preclinical evidence demonstrates that Nilotinib, a multi-kinase tyrosine kinase inhibitor, increases autophagic clearance of neurotoxic proteins and ameliorates other neurodegenerative pathologies. In humans we have shown that Nilotinib penetrates the brain, inhibits cerebrospinal fluid (CSF) Abelson tyrosine kinase activity, reduces CSF total and phosphorylated tau, stabilizes CSF alpha-synuclein levels, and increases CSF Homovanillic acid, an end byproduct of Dopamine metabolism. MicroRNAs (miRNAs) have recently emerged as attractive candidates as biomarkers in neurodegenerative diseases due to their stability in biological fluids including CSF and plasma. MiRNA are small non-coding RNA whose role is to post-translationally silence expression of gene targets. A pilot study identified 47 miRNA in the CSF of PD and Dementia with Lewy Bodies (DBL) patients differentially expressed after 6 months Nilotinib treatment compared to baseline. Target analysis indicates that many of the down regulated miRNA target genes are associated with autophagy and ubiquitination, agreeing with much of our preclinical work that demonstrates autophagy and ubiquitination leading to proteolysis is inhibited in PD and disinhibited after Nilotinib treatment. Here, I propose to further evaluate the effects of TKI via Nilotinib on gene regulation by sequencing the miRNA in CSF collected from 50 patients enrolled in NCT02954978 to evaluate Nilotinib’s impact in PD and 37 patients enrolled in NCT02954978 to evaluate Nilotinib’s impact in AD. Next- generation sequencing will identify miRNAs that were differentially expressed in Nilotinib treated individuals compared to baseline. Next-generation sequencing will be validated using TaqMan real-time PCR assays. Gene target analysis will be performed to reveal what genes and pathways are associated with the expressed miRNAs. Surrogate protein biomarkers for PD progression will be measured in the CSF using enzyme-linked immunosorbent assays (ELISAs). Together, this wealth of collected data will enable me to correlate miRNA changes to clinical endpoint measures including protein biomarkers and clinical rating scale scores to validate miRNAs as potential reliable biomarkers for the response to TKI via Nilotinib in PD and AD.

项目摘要/摘要： 阿尔茨海默氏病（AD）和帕金森氏病（PD）估计会影响曼联的40％和1％ 规定的人口分别超过85岁，使其成为两个最常见的神经退行性 疾病[1-6]。可靠的生物标志物识别用于响应AD和PD干预措施的可靠生物标志物 协助开发成功修饰疗法的疾病。我们有力的临床前证据 证明尼洛替尼是一种多激酶酪氨酸激酶抑制剂，增加了神经毒性的自噬清除率 蛋白质并改善其他神经退行性病理。在人类中，我们已经表明了尼洛蒂尼 穿透大脑，抑制脑脊液（CSF）阿伯森酪氨酸激酶活性，减少CSF的总和 磷酸化的tau，稳定CSFα-突触核蛋白水平，并增加CSF同型甲酸，末端 多巴胺代谢的副产品。 microRNA（mirnas）最近出现了作为吸引人的候选人 神经退行性疾病中的生物标志物在包括CSF和血浆在内的生物液中稳定性。 miRNA是小的非编码RNA，其作用是基因靶标的翻译后沉默表达。飞行员 研究在PD的CSF和痴呆症患者（DBL）患者的CSF中鉴定了47个miRNA 与基线相比，尼洛替尼治疗6个月后表示。目标分析表明许多 下调的miRNA靶基因与自噬和泛素化有关，与大部分 PD和 尼洛替尼治疗后抑制。在这里，我建议进一步评估TKI通过Nilotinib对基因的影响 通过对从NCT02954978中招募的50名患者收集的CSF进行测序来调节MIRNA来评估 Nilotinib对PD的影响和37例NCT02954978招收的患者，以评估Nilotinib在AD中的影响。下一个- 生成测序将识别在尼洛替尼治疗个体中不同表达的miRNA 与基线相比。下一代测序将使用Taqman实时PCR分析验证。基因 将进行目标分析以揭示哪些基因和途径与表达的miRNA相关。 PD进展的替代蛋白生物标志物将在CSF中使用酶连接在CSF中测量 免疫吸附测定法（ELISA）。这些收集的数据共同使我能够关联mirna 更改临床终点测量值，包括蛋白质生物标志物和临床评级量表，以验证 miRNA是通过Nilotinib在PD和AD中对TKI反应的潜在可靠生物标志物。