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.

项目概要/摘要： 据估计，阿尔茨海默病（AD）和帕金森病（PD）分别影响40%和1%的美国人。 分别声明85岁以上人群，使他们成为两种最常见的神经退行性疾病 疾病[1 - 6]识别一种可靠的生物标志物，用于AD和PD干预措施的应答， 帮助开发成功的疾病修饰疗法。我们强大的临床前证据 表明尼洛替尼，一种多激酶酪氨酸激酶抑制剂， 蛋白质和改善其他神经退行性病变。在人类中，我们已经证明尼洛替尼 穿透大脑，抑制脑脊液（CSF）Abelson酪氨酸激酶活性，减少CSF总量， 磷酸化tau，稳定CSF α-突触核蛋白水平，并增加CSF高香草酸， 多巴胺代谢的副产品。microRNAs（miRNAs）最近成为有吸引力的候选者， 这些生物标志物在神经退行性疾病中的应用是由于它们在生物流体（包括CSF和血浆）中的稳定性。 miRNA是一种小的非编码RNA，其作用是使靶基因的表达在转录后沉默。一个试点 一项研究在PD和路易体痴呆（DBL）患者的CSF中鉴别出47种差异miRNA 与基线相比，尼洛替尼治疗6个月后表达。目标分析表明， 下调的miRNA靶基因与自噬和泛素化有关，这与许多研究结果一致。 我们的临床前工作表明，导致蛋白水解的自噬和泛素化在PD中受到抑制， 尼洛替尼治疗后解除抑制。在此，我建议进一步评估TKI通过尼洛替尼对基因表达的影响。 通过对从入组NCT 02954978的50名患者收集的CSF中的miRNA进行测序来进行调节，以评估 尼洛替尼对PD的影响和37例患者入组NCT 02954978，以评价尼洛替尼对AD的影响。下一篇： 世代测序将鉴定尼洛替尼治疗个体中差异表达的miRNA 与基线相比。下一代测序将使用TaqMan实时PCR检测进行验证。基因 将进行靶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使用酶联免疫吸附法在CSF中测量。 免疫吸附测定（ELISA）。总之，这些收集到的丰富数据将使我能够将miRNA 变更临床终点指标，包括蛋白质生物标志物和临床评定量表评分，以验证 miRNA作为PD和AD患者通过尼洛替尼对TKI应答的潜在可靠生物标志物。