Investigation of urinary extracellular vesicles as novel and safe therapeutics for autosomal recessive polycystic kidney disease

尿细胞外囊泡作为常染色体隐性遗传性多囊肾病的新型安全疗法的研究

• 批准号: 10750704
• 负责人: Eun Ji Chung
• 金额: $ 46.57万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-04 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750704
• 关键词: Address; Adult; Affect; Age; Amniotic Fluid; Autosomal Dominant Polycystic Kidney; Autosomal Recessive Polycystic Kidney; Binding; Biodistribution; Biological; Birth; Cell Communication; Cells; Cessation of life; Characteristics; Childhood; Clinical Trials; Development; Disease; Disease Progression; Dose; Drug Kinetics; Drug Side Effects; End stage renal failure; FDA approved; Fetal Lung; Fetus; Genetic; Half-Life; Hepatotoxicity; Home; Hour; Human; In Vitro; Injections; Intravenous; Investigation; Kidney; Kidney Diseases; Kidney Failure; Kidney Transplantation; Life; Live Birth; Measures; Membrane; Messenger RNA; Modeling; Mothers; Mus; Mutate; Mutation; Nephrology; Newborn Infant; Oligohydramnios; PKHD1 gene; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Pilot Projects; Polycystic Kidney Diseases; Proliferating; Property; Proteins; Pulmonary Valve Insufficiency; RNA; Rattus; Receptor Protein-Tyrosine Kinases; Renal function; Research; Respiration; Safety; Source; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Translating; Treatment Efficacy; Tyrosine Kinase Inhibitor; Urine; Vasopressin Receptor; Work; autosome; biomaterial compatibility; body cavity; congenital hepatic fibrosis; cpk mouse; design; disease phenotype; effective therapy; exosome; extracellular vesicles; fetal; gene product; gene therapy; in utero; in vivo; inhibitor; insight; kidney cell; lung development; mouse model; multidisciplinary; nanomedicine; nanoparticle; neonate; novel; novel strategies; novel therapeutic intervention; novel therapeutics; patient population; pediatric patients; pregnant; protein expression; pulmonary hypoplasia; pup; small molecule; targeted treatment; therapeutic nanoparticles; therapy development; therapy outcome; tolvaptan; urinary

ABSTRACT Autosomal recessive polycystic kidney disease (ARPKD) is a leading cause of kidney failure in childhood and is caused primarily by mutations in the PKHD1 gene as well as mutations in CYS1. Although ARPKD patients progress to end stage kidney disease at varying ages, 20-30% of ARPKD patients have severe disease which results in death 24-48 hours after birth. Despite the tremendous unmet need, limited progress has been made for ARPKD treatment and no FDA-approved therapies are available for ARPKD patients. Although tolvaptan, a small molecule drug that was approved in 2018 for autosomal dominant PKD which mostly affects adults, is currently under phase I clinical trials for ARPKD, tolvaptan is not kidney-targeted and results in off-target effects. Additionally, tolvaptan’s long-term efficacy, tolerability, and developmental consequences in a primarily pediatric ARPKD patient population and its ability to enhance survival in neonates with severe ARPKD is unknown. Thus, new therapeutic strategies that can address both safety and efficacy in ARPKD are urgently needed. To that end, the purpose of this proposal is to examine the potential of urinary extracellular vesicles (uEVs) as a novel and safe therapy for ARPKD. EVs, also known as exosomes, are secreted, membrane-bound, biological nanoparticles that facilitate cell-to-cell communication and contain RNA and protein cargo characteristic to their parent cell. We propose uEVs for ARPKD therapy for several reasons: uEVs are 1) inherently biocompatible which is critically important when developing therapies for pediatric patients, 2) carry fibrocystin and cystin, the gene products of PKHD1 and CYS1 which are mutated in ARPKD, respectively, 3) have been found to home to the kidneys and transfer functional proteins to induce a therapeutic outcome in kidney disease, and 4) obtaining high quantities uEV from urine, which is normally discarded, is feasible, noninvasive, and cheap. We hypothesize that uEVs derived from non-disease sources are a safe therapy that can be used to deliver and supplement functional proteins including fibrocystin and cystin that are defective in ARPKD to inhibit disease progression. To test our hypothesis, we will first characterize the nanoparticle properties of uEVs and evaluate uEV fibrocystin and cystin protein and mRNA cargo, cell internalization, and therapeutic effects in renal cells in vitro (Aim 1). Next, we will administer uEVs intravenously in slowly progressing and severe ARPKD murine models and evaluate the pharmacokinetic properties, therapeutic efficacy, and safety in vivo (Aim 2.1). Finally, given most human ARPKD leads to death in utero or in newborns shortly after birth, we will deliver uEVs in utero in pregnant mice and evaluate the ability to extend survival in severe ARPKD upon fetal delivery (Aim 2.2). Through our multidisciplinary, investigative team of nanomedicine and pediatric nephrology, we are well-equipped and fully committed to successfully carry out this work that is much needed in ARPKD therapy.

抽象的 常染色体隐性多囊肾病 (ARPKD) 是儿童期肾衰竭的主要原因， 主要由 PKHD1 基因突变和 CYS1 突变引起。尽管 ARPKD 患者 在不同年龄进展为终末期肾病，20-30%的 ARPKD 患者患有严重疾病， 导致出生后 24-48 小时内死亡。尽管存在巨大的未满足需求，但取得的进展有限 ARPKD 治疗目前尚无 FDA 批准的治疗方法可用于 ARPKD 患者。尽管托伐普坦 2018 年批准用于治疗主要影响成人的常染色体显性多囊肾病的小分子药物是 目前正在进行针对 ARPKD 的 I 期临床试验，托伐普坦不是肾脏靶向的，会导致脱靶效应。 此外，托伐普坦在主要儿科患者中的长期疗效、耐受性和发育后果 ARPKD 患者群体及其提高严重 ARPKD 新生儿生存率的能力尚不清楚。因此， 迫切需要能够解决 ARPKD 安全性和有效性的新治疗策略。对此 最后，该提案的目的是检查尿液细胞外囊泡（uEV）作为一种新型药物的潜力 ARPKD 的安全治疗。 EV，也称为外泌体，是分泌的、膜结合的、生物性的 纳米粒子促进细胞间通讯并含有其特有的RNA和蛋白质货物 父细胞。我们建议 uEV 用于 ARPKD 治疗有几个原因：uEV 1) 本质上具有生物相容性 这在为儿科患者开发治疗方法时至关重要，2) 携带纤维囊肿蛋白和胱氨酸， PKHD1 和 CYS1 的基因产物分别在 ARPKD 中发生突变，3) 已被发现 肾脏并转移功能蛋白以诱导肾脏疾病的治疗结果，以及 4) 获得 从通常被丢弃的尿液中提取大量 uEV 是可行的、非侵入性的且便宜。我们假设 来自非疾病来源的 uEV 是一种安全的疗法，可用于传递和补充 ARPKD 中存在缺陷的功能蛋白（包括纤维囊肿蛋白和胱氨酸）可抑制疾病进展。到 为了检验我们的假设，我们将首先表征 uEV 的纳米颗粒特性并评估 uEV 纤维囊肿蛋白 胱氨酸蛋白和 mRNA 货物、细胞内化以及体外肾细胞的治疗效果（目标 1）。 接下来，我们将在进展缓慢和严重的 ARPKD 小鼠模型中静脉注射 uEV， 评估体内药代动力学特性、治疗功效和安全性（目标 2.1）。最后，给出最 人类 ARPKD 会导致子宫内或新生儿出生后不久死亡，我们将在怀孕期间在子宫内输送 uEV 小鼠并评估在胎儿分娩时延长严重 ARPKD 患者生存的能力（目标 2.2）。通过我们的 纳米医学和小儿肾脏病学多学科、研究团队，设备齐全，设施齐全 致力于成功开展 ARPKD 治疗急需的这项工作。