MicroRNA-Based Therapeutics for Rare Cystic Kidney Diseases

基于 MicroRNA 的罕见囊性肾病治疗方法

• 批准号: 7936337
• 负责人: Peter Igarashi
• 金额: $ 48.61万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936337
• 关键词: Adult; Affect; Animal Model; Animals; Autosomal Recessive Polycystic Kidney; Binding; Biological Process; Blindness; Cardiovascular Diseases; Cells; Child; Clinical; Clinical Data; Clinical Trials; Code; Cyst; Cystic Kidney Diseases; Cystic kidney; Development; Diabetes Mellitus; Disease; End stage renal failure; Family; Fibrosis; Functional RNA; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Goals; Hereditary Disease; Human; Incidence; Individual; Infant; Kidney; Kidney Diseases; Kidney Failure; Knockout Mice; Laboratories; Link; Liquid substance; Liver Fibrosis; Malignant Neoplasms; Messenger RNA; Methods; MicroRNAs; Microarray Analysis; Modeling; Molecular; Molecular Profiling; Molecular Target; Morbidity - disease rate; Mus; Mutant Strains Mice; Mutate; Mutation; Nephronophthisis; Newborn Infant; Normal Range; Oligonucleotides; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Plants; Play; Principal Investigator; Proteins; Rare Diseases; Regulator Genes; Renal function; Role; Sequence Analysis; Staging; Testing; Therapeutic; Tissues; Transcript; Translations; Virus Diseases; base; effective therapy; human disease; improved; in vivo; mRNA Transcript Degradation; mortality; mouse model; mutant; nervous system disorder; new technology; novel; novel strategies; novel therapeutic intervention; overexpression; pre-clinical; programs; public health relevance; restoration; transcription factor

DESCRIPTION (provided by applicant): The goal of this two-year proposal is to test the feasibility of microRNA-based therapeutics as a novel strategy for the treatment of rare cystic kidney diseases. Autosomal recessive cystic kidney diseases include autosomal recessive polycystic kidney disease (ARPKD), renal cysts and diabetes (RCAD), and nephronophthisis (NPHP). These rare genetic disorders are characterized by the formation of cysts in the kidneys and progressive kidney failure. Extrarenal manifestations include congenital hepatic fibrosis (ARPKD), diabetes (RCAD), and blindness (NPHP). Although these disorders are rare with incidences of 1:20,000 or less, they have significant clinical importance. ARPKD and NPHP are the most common monogenic causes of end-stage kidney disease in infants and children. The genes that are mutated in human ARPKD, RCAD, and NPHP have been identified. However, no specific and effective therapies exist for these diseases, and no drugs are currently in clinical trials. Therefore, new therapeutic approaches are needed. For the past 10 years, our laboratory has investigated the molecular pathogenesis of autosomal recessive cystic kidney diseases. We have developed orthologous mouse models carrying mutations in the same genes that are mutated in humans with these disorders. The phenotypes of the mutant mice resemble the phenotypes of affected humans indicating that they represent excellent animal models. Analysis of the mutant mice has helped unravel the pathogenesis of the human diseases and revealed that they are linked in a common pathway. In particular, the transcription factor HNF-1¿, which is mutated in RCAD, regulates the transcription of PKHD1, which is mutated in ARPKD, and GLIS2 and NPHP3, which are mutated in nephronophthisis. Thus, correction of abnormalities in this pathway may be an effective therapeutic approach for cystic kidney diseases. Recent studies have identified microRNAs (miRNAs) as potential molecular targets in human diseases. miRNAs are short, non-coding RNAs that regulate post-transcriptional gene expression by inhibiting translation or promoting cleavage of complementary messenger RNAs. In preliminary studies, we have identified several families of miRNAs that are abnormally overexpressed in HNF-1¿ mutant cells. The sequences of the miRNAs are complementary to mRNAs encoded by known cystic disease genes, such as PKD2 and PKHD1. These studies identify a novel role of miRNAs in the pathogenesis of cystic kidney diseases and suggest that inhibition of miRNAs may be an effective strategy for reducing cyst formation. Recently, effective methods for modulating the activity of miRNAs in vivo have been developed. Antagomirs are chemically modified oligonucleotides that can be administered parenterally and that specifically and durably inactivate target miRNAs. In this proposal, we will use miRNA microarray analysis to comprehensively identify miRNAs that are abnormally expressed in orthologous mouse models of ARPKD, RCAD, and NPHP. Next, we will test whether the administration of antagomirs targeting the overexpressed miRNAs inhibit cyst formation and improve renal function in mutant mice. Such studies would provide important pre-clinical data demonstrating the feasibility of this approach for the treatment of these rare but clinically important genetic disorders and would also have broad implications for the treatment of other rare disorders affecting the kidney. PUBLIC HEALTH RELEVANCE: Autosomal recessive cystic kidney diseases are rare genetic disorders that produce kidney failure in children. No specific treatment currently exists. This proposal will test the feasibility of treating these disorders using a new technology, microRNA-based therapeutics.

描述（由申请人提供）：这项为期两年的提案的目标是测试基于microRNA的疗法作为治疗罕见囊性肾病的新策略的可行性。常染色体隐性遗传性囊性肾病包括常染色体隐性遗传性多囊肾病（ARPKD）、肾囊肿和糖尿病（RCAD）以及肾单位结核（NPHP）。这些罕见的遗传性疾病的特点是形成囊肿的肾脏和进行性肾衰竭。肾外表现包括先天性肝纤维化（ARPKD）、糖尿病（RCAD）和失明（NPHP）。虽然这些疾病的发病率为1：20，000或更低，但它们具有重要的临床意义。ARPKD和NPHP是婴儿和儿童终末期肾病最常见的单基因病因。在人类ARPKD、RCAD和NPHP中突变的基因已经被鉴定。然而，对于这些疾病没有特异性和有效的治疗方法，目前也没有药物在临床试验中。因此，需要新的治疗方法。在过去的10年里，我们的实验室研究了常染色体隐性遗传性囊性肾病的分子发病机制。我们已经开发了携带突变基因的小鼠模型，这些突变基因在患有这些疾病的人类中发生突变。突变小鼠的表型与受影响人类的表型相似，表明它们代表了极好的动物模型。对突变小鼠的分析有助于揭示人类疾病的发病机制，并揭示它们在一个共同的途径中相互联系。特别是，在RCAD中突变的转录因子HNF-1？调节在ARPKD中突变的PKHD 1以及在肾单位肾结核中突变的GLIS 2和NPHP 3的转录。因此，纠正这一途径的异常可能是治疗囊性肾病的有效方法。最近的研究已经确定microRNAs（miRNAs）作为人类疾病的潜在分子靶点。miRNA是短的非编码RNA，其通过抑制互补信使RNA的翻译或促进互补信使RNA的切割来调节转录后基因表达。在初步的研究中，我们已经确定了几个家族的miRNAs异常过表达的HNF-1突变细胞。miRNA的序列与已知囊性疾病基因（如PKD 2和PKHD 1）编码的mRNA互补。这些研究确定了miRNAs在囊性肾病发病机制中的新作用，并表明抑制miRNAs可能是减少囊肿形成的有效策略。最近，已经开发了用于在体内调节miRNA活性的有效方法。拮抗剂是化学修饰的寡核苷酸，其可以肠胃外施用，并且特异性且持久地靶向miRNA。在这个提议中，我们将使用miRNA微阵列分析来全面鉴定在ARPKD、RCAD和NPHP的正向小鼠模型中异常表达的miRNA。接下来，我们将测试靶向过表达的miRNAs的西洛米的施用是否抑制突变小鼠中的囊肿形成并改善肾功能。这些研究将提供重要的临床前数据，证明这种方法治疗这些罕见但具有临床重要性的遗传性疾病的可行性，并对治疗影响肾脏的其他罕见疾病具有广泛的意义。 公共卫生相关性：常染色体隐性遗传性囊性肾病是一种罕见的遗传性疾病，可导致儿童肾衰竭。目前没有具体的治疗方法。该提案将测试使用新技术，基于microRNA的疗法治疗这些疾病的可行性。