Preventing hyperphagia in Prader Willi syndrome using an oligonucleotide

使用寡核苷酸预防普瑞德威利综合征的食欲过盛

基本信息

批准号：
8824160
负责人：
Stefan Stamm
金额：
$ 22.51万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-23 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8824160
关键词：
Acute Address Adverse effects Anti-Obesity Agents Area Behavioral Blood Blood - brain barrier anatomy Blood Circulation Brain Brain region Canis familiaris Carotid Arteries Cell membrane Cell surface Cells Chemistry Child Comorbidity Detection Disease Drug Targeting Eating Endoplasmic Reticulum Exons FOS gene Fasting Food Functional RNA Genetic Genetic Transcription Genome Goals Health Heterodimerization Hyperphagia Hypothalamic structure Immune response Inflammation Injection of therapeutic agent Insulin Investigation Leptin Measures Messenger RNA Modeling Morbid Obesity Mus Neurologic Neurons Nucleic Acids Obesity Obsession Oligonucleotides Patients Pharmaceutical Preparations Pharmacodynamics Prader-Willi Syndrome Primates Pro-Opiomelanocortin Problem behavior Protein Isoforms Proteins Pump RNA RNA Processing RNA Sequences RNA Splicing Reagent Safety Satiation Serotonin Serotonin Receptor 5-HT2C Signal Transduction Site Small Nucleolar RNA Structure Structure of nucleus infundibularis hypothalami System Techniques Testing Therapeutic Time Toxic effect United States Work base ghrelin innovation insight mRNA Precursor nervous system disorder non-genetic prevent public health relevance receptor response tool uptake

项目摘要

DESCRIPTION (provided by applicant): One of the most severe daily problems in Prader-Willi syndrome (PWS) is the inability to reach satiety after a meal, which leads to hyperphagia and obesity. The loss of expression of two small nucleolar RNA (snoRNA) clusters is a crucial genetic contributor to the disease. One of these snoRNAs (SNORD115) promotes the most active form of the serotonin receptor 2C (HTR2C). Activated HTR2C receptors in the arcuate nucleus inhibit food uptake. We developed an oligonucleotide ("oligo#5") that similar to SNORD115 promotes the formation of the most active HTR2C receptor that localizes on the cell surface. Oligo#5 inhibits food uptake of fasted mice when delivered by intracerebroventricular (ICV) or carotid injection to the arcuate nucleus. We postulate that oligo#5 could be developed into an anti hyperphagia drug for Prader-Willi syndrome. This hypothesis will be tested in two Specific Aims: 1. Deliver oligo#5 and its derivatives to the arcuate nucleus in the brain and determine their mode of action. We will test delivery using osmotic pumps bringing oligo#5 into the 3rd ventricle, similar to the acute injections. Oligo#5 accumulates in the hypothalamus and decreases food uptake after carotid injection, suggesting it can cross the blood brain barrier. We will therefore test delivery by injecting into the bloodstream. Different chemistries of oligo#5 wil be tested in cell-based models prior to testing in mice. This allows to determine efficacy and to get mechanistic insights. 2. Determine long term toxicity, efficacy and off-site targets of the oligonucleotide. We did not observe adverse effects in up to three repeated injections and will test long-term delivery using osmotic pumps and injection into the blood. We will determine immune response, behavioral changes, inflammation and other adverse effects and measure changes in efficacy over time. Off-target effects will be measured by RNA sequencing and array analysis. We expect that oligo#5 is safe, can be delivered through the bloodstream and shows a long-term reduction in food intake. These investigations are significant, as they target hyperphagia, the most complicating feature in the daily lives of patients, they are innovative as they deliver a nucleic acid to a brain region possibly through the blood, where it acts by changing target RNA processing; they are feasible, as we developed the oligonucleotide and the detection system.

描述（由申请人提供）：Prader-Willi综合征（PWS）中最严重的每日问题之一是进餐后无法达到饱腹感，这导致了肥大和肥胖。两个小核仁RNA（SNORNA）簇的表达丧失是对该疾病的关键遗传贡献。这些snornas之一（SNORD115）促进了5-羟色胺受体2C（HTR2C）的最活跃形式。弧形核中活化的HTR2C受体抑制食物的吸收。我们开发了一种与SNORD115相似的寡核苷酸（“ Oligo＃5”），促进了定位在细胞表面上的最活跃的HTR2C受体的形成。寡核＃5抑制禁食小鼠的食物吸收，当时脑室内（ICV）或颈动脉注射向弧形核。我们假设寡核＃5可以发展成为用于普拉德 - 威利综合征的抗多晶状体药物。该假设将以两个具体的目的进行检验：1。将寡核＃5及其衍生物传递到大脑中的弧形核并确定其作用方式。我们将使用渗透泵测试输送，将Oligo＃5带入第三个心室，类似于急性注射。寡核＃5在下丘脑中积聚，并减少颈动脉注射后的食物摄取，这表明它可以越过血脑屏障。因此，我们将通过注射血液来测试递送。在小鼠进行测试之前，将在基于细胞的模型中对寡核酸的不同化学分配进行测试。这允许确定功效并获得机理见解。 2。确定寡核苷酸的长期毒性，功效和异地靶标。我们没有观察到多达三次重复注射的不良反应，并且将使用渗透泵和注射到血液中进行长期递送。我们将确定免疫反应，行为变化，炎症和其他不良反应，并衡量随着时间的推移疗效的变化。脱靶效应将通过RNA测序和阵列分析来衡量。我们预计唯一的寡核糖是安全的，可以通过血液传递，并长期降低食物摄入量。这些研究很重要，因为它们瞄准了患者日常生活中最复杂的特征，因此它们具有创新性，因为它们可能通过血液向大脑区域运送核酸，在该血液中，它通过改变靶标RNA处理而起作用。当我们开发寡核苷酸和检测系统时，它们是可行的。