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

基本信息

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
项目类别：
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

项目摘要

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.

研究总结非编码RNA（lncRNA）在人类遗传疾病中的功能作用和分子机制成为有希望的新领域，为未来的研究方向和治疗奠定基础目前缺乏足够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，导致其酶活性中断，从而导致血液中苯丙氨酸的积累。我们的初步数据表明，人类 lncRNA HULC 在功能上与小鼠 lncRNA 保守一对。 PKU 患者携带 HULC 基因的基因组变异。一对的敲除会导致色素沉着不足，生长迟缓、进行性神经系统症状和癫痫发作，忠实地模拟了人类 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，它们是不可分割的建立lncRNA-酶相互作用并设计肽标记的lncRNA模拟物来恢复 PAH 的酶活性。在 PKU 小鼠模型中施用 HULC 模拟物成功缓解了过量血清中的苯丙氨酸水平，提供对遗传性遗传疾病基础的机制洞察。这拟议的研究旨在建立长非编码 RNA 作为起始和进展的重要参与者先天性代谢紊乱的研究并开发基于 lncRNA 的创新方法来确定新策略解决 PKU 的分子基础。中心假设是lncRNA的基因突变和缺失导致酶促 PAH 缺乏，可以通过靶向治疗增强蛋白质稳定性和 PAH 的酶活性。使用源自 PKU 患者的 iPS 分化肝细胞，我们首先通过解决以下问题，证明 HULC 在 PKU 发生和进展中的生物学意义常见 HULC 突变的影响以及评估 HULC 突变或缺失与苯丙酮尿症症状。然后我们将阐明lncRNA介导的PAH调节的分子机制酶活性。最后，我们将使用富含肝脏的 lncRNA 模拟物和小分子激动剂/拮抗剂以确定其在恢复受损酶功能中的机制作用。上述研究目标不仅将阐明lncRNA在代谢紊乱中的作用，同时也说明了lncRNA模拟物和小分子抑制剂药物的潜在治疗价值在治疗先天性遗传疾病方面。我们预计这项研究的结果将改变当前的研究和临床范式结合非编码基因的生理和病理方面，并为未来基于 lncRNA 的药物的开发之路。