Molecular Mechanism of long Noncoding RNAs in Phenylketonuria

长非编码RNA在苯丙酮尿症中的分子机制

• 批准号: 10562363
• 负责人: VERNON R SUTTON
• 金额: $ 50.39万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-10 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10562363
• 关键词: Address; Agonist; Alleles; Animals; Biological; Biological Markers; Blood; Categories; Chromatin; Classical phenylketonuria; Clinical; Code; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; DNA Sequence Alteration; Data; Development; Diet; Disease; Enhancers; Enzyme Interaction; Enzymes; Event; Female; Foundations; Frequencies; Future; Genes; Genetic; Genetic Diseases; Genetic Transcription; Goals; Growth; Hepatocyte; Hereditary Disease; Human; Human Genetics; Hydrolysis; Hypopigmentation; Impairment; Indole-3-Carbinol; Inherited; Knock-out; Lead; Liver; Lymphokine-Activated Killer Cells; Mediating; Medical Research; Medicine; Mental Retardation; Metabolic; Metabolic Diseases; Metabolism; Molecular; Mus; Mutation; Neurologic Symptoms; Neurons; Odors; Oligonucleotides; Outcome; Pathologic; Patients; Peptides; Phenylalanine; Phenylalanine Hydroxylase; Phenylketonurias; Phosphorylation; Physiological; Plasma; Polyubiquitination; Post-Translational Protein Processing; Process; Protein Kinase; Proteins; Publishing; RNA; Regulation; Research; Role; Science; Seizures; Serum; Severities; Signal Transduction; Supplementation; Symptoms; System; Therapeutic; Tyrosine; Untranslated RNA; Variant; Work; antagonist; design; dietary; frontier; genetic variant; human model; improved; induced pluripotent stem cell; inhibitor; innovation; insight; male; mouse model; mutant; precision medicine; protein degradation; small molecule; small molecule inhibitor; targeted treatment; therapeutic RNA; treatment strategy; ubiquitin-protein ligase

Research Summary The functional roles and molecular mechanisms of non-coding RNAs (lncRNA) in human genetic disease serve as promising new frontiers for establishing the foundation of future research directions and therapeutic avenues for disorders that currently lack sufficient treatment options. The inborn metabolic disorder phenylketonuria (PKU) has been considered to be caused by mutations in the phenylalanine hydroxylase (PAH) gene, resulting in disruptions to its enzymatic activity and consequent accumulation of phenylalanine in the blood. Our preliminary data demonstrated that the human lncRNA HULC is functionally conserved with mouse lncRNA Pair. PKU patients harbor genomic variants of the HULC gene. Knockout of Pair leads to hypo-pigmentation, growth retardation, progressive neurological symptoms, and seizures, which faithfully models human PKU. Deletion of HULC in primary human hepatocytes leads to elevated cellular Phenylalanine, suggesting the functional importance of HULC/Pair in PKU. We defined the RNA motifs of both HULC and Pair that are integral to establishing lncRNA-enzyme interactions and designed peptide-tagged lncRNA mimics that restored the enzymatic activity of PAH. Administration of HULC mimics in PKU mouse model successfully alleviated excess phenylalanine levels in serum, providing mechanistic insight into the basis of inherited genetic diseases. The proposed study seeks to establish long non-coding RNAs as important players in the initiation and progression of inborn metabolic disorders and develop a lncRNA-based innovative approach for determining new strategies to tackle the molecular basis of PKU. The central hypothesis is that the genetic mutation and deletion of lncRNA results in the enzymatic deficiency of PAH, which could be alleviated through targeting therapies to enhance the protein stability and enzymatic activity of PAH. Using iPS-differentiated hepatocytes derived from PKU patients, we will first demonstrate the biological significance of HULC in the development and progression of PKU by addressing the impact of common HULC mutations and assessing the correlations between mutations or deletions of HULC and PKU symptoms. We will then elucidate the molecular mechanisms of lncRNA-mediated regulation of PAH enzymatic activity. Finally, we will use the administration of liver-enriched lncRNA mimics and small molecule agonists/antagonists to establish their mechanistic role in restoring impaired enzymatic function. The aforementioned research aims will not only elucidate the roles of lncRNAs in metabolic disorders, but also illustrate the potential therapeutic value of lncRNA mimics and small molecule inhibitor-based medicine in treating inborn genetic diseases. We anticipate that the results of this study will shift the current research and clinical paradigm to incorporate the physiological and pathological aspects of non-coding genes and pave the way for the development of future lncRNA-based medicines.

研究综述 非编码RNA（lncRNA）在人类遗传性疾病中的作用及其分子机制 作为建立未来研究方向和治疗基础的有前途的新前沿， 目前缺乏足够的治疗选择的疾病的途径。先天性代谢紊乱 苯丙酮尿症（PKU）被认为是由苯丙氨酸羟化酶（PAH）突变引起的 基因，导致其酶活性的破坏和随之而来的苯丙氨酸在血液中的积累。 我们的初步数据表明，人类lncRNA HULC与小鼠lncRNA HULC在功能上是保守的。 汇率PKU患者携带HULC基因的基因组变体。敲除Pair导致色素沉着减退， 生长迟缓、进行性神经症状和癫痫发作，其忠实地模拟了人PKU。 原代人肝细胞中HULC的缺失导致细胞苯丙氨酸升高，这表明 HULC/Pair在PKU中的功能重要性。我们定义了HULC和Pair的RNA基序， 建立lncRNA-酶相互作用，并设计了肽标记的lncRNA模拟物， PAH的酶活性。在PKU小鼠模型中施用HULC模拟物成功地减轻了过量的 血清中的苯丙氨酸水平，为遗传性遗传疾病的基础提供了机制性的见解。的 一项拟议的研究旨在建立长非编码RNA作为启动和进展的重要参与者， 先天性代谢紊乱，并开发一种基于lncRNA的创新方法， 来解决PKU的分子基础。 中心假设是lncRNA的基因突变和缺失导致酶促降解。 多环芳烃缺乏，可以通过靶向治疗来缓解，以增强蛋白质稳定性， PAH的酶活性。使用来自PKU患者的iPS分化的肝细胞，我们将首先 通过解决以下问题，证明HULC在PKU发展和进展中的生物学意义： 常见HULC突变的影响以及评估HULC突变或缺失与 PKU症状。然后，我们将阐明lncRNA介导的PAH调节的分子机制， 酶活性最后，我们将使用肝脏富集的lncRNA模拟物和小分子 激动剂/拮抗剂以确定它们在恢复受损的酶功能中的机制作用。 上述研究目的不仅将阐明lncRNA在代谢紊乱中的作用， 而且还说明了lncRNA模拟物和基于小分子通道的药物的潜在治疗价值 治疗先天性遗传疾病的方法我们预计，这项研究的结果将改变目前的研究， 临床范例，以纳入非编码基因的生理和病理方面， 为未来基于lncRNA的药物的开发提供了一条途径。