Development of self-delivering RNAi to evaluate PTEN as a therapeutic target to p

开发自传递 RNAi 来评估 PTEN 作为 p 治疗靶点

基本信息

批准号：
8645823
负责人：
Lisa J McKerracher
金额：
$ 22.48万
依托单位：
BIOAXONE BIOSCIENCES, INC.
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：
2014-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8645823
关键词：
Aftercare Animals Axon Biochemical Cell Culture Techniques Cell Survival Cells Chemicals Clinical Clinical Chemistry Combination Drug Therapy Dermal Development Dose Double-Stranded RNA EGF gene Ensure Environment Gene Silencing Genes Glioblastoma Goals Growth Hela Cells Human In Vitro Inflammation Injury Label Lead Learning Length Lysine Mammalian Cell Measurement Measures Mediating Methods Modeling Modification Molecular Morphology Natural regeneration Nerve Crush Neuraxis Neurites Neuroglia Neuronal Differentiation Neurons Operative Surgical Procedures Optic Nerve Outcome PC12 Cells PTEN gene Pharmaceutical Preparations Proteins RNA RNA Interference RNA Sequences Recombinants Recovery Recovery of Function Reporter Research Retina Retinal Retinal Ganglion Cells Risk Rodent Safety Signal Transduction Silicon Dioxide Small Interfering RNA Spinal Cord Spinal cord injury System Technology Testing Therapeutic Therapeutic Uses Toxic effect Transgenic Mice Translating Viral Vector Western Blotting Work axon regeneration axonal sprouting central nervous system injury design drug candidate drug development efficacy testing experience improved in vitro testing in vivo injured innovation knock-down neurite growth novel programs public health relevance regenerative repaired research study response therapeutic target tumor

项目摘要

Project Summary Innovative drug/therapy combinations directed at multiple and proven therapeutic targets have the potential to dramatically improve outcomes after SCI. Studies on axon regeneration and recovery in rodents have revealed that the spinal cord is not hard wired and "learning" can occur in spinal cord circuits. Compounds that elicit axonal regeneration and sprouting help plasticity in the spinal cord and dramatically improve recovery from traumatic injury, at least in rodents. Many different compounds/therapies tested in rodents promote regeneration. Almost without exception, compounds that promote regeneration also improve functional recovery after spinal cord injury. Recently, very impressive axon regeneration was obtained from deletion of PTEN. The goal of this application is to simultaneously evaluate PTEN as target for therapeutic treatment of spinal cord injury and a novel RNAi delivery technology to provide prolonged in vivo gene knockdown effect. We will create "self-delivering" small interfering RNAs (sdRNA) targeting PTEN. The chemical modification introduced into sdRNA molecules makes them cell- permeable. This technology has been demonstrated to work in vivo in several applications, including delivery to the retina. As the first step we will synthetize a panel of sdRNA sequences and evaluate them in mammalian cell cultures to select lead candidate(s) efficient in PTEN knockdown. Next, we will test the compounds in neuron cell cultures, then with primary neurons. We will also examine effects on morphology and proliferation of primary glial cells and human glioblastoma cells. Once we determine lead candidate(s) that promote robust neurite growth, we will test the compounds for efficacy of knockdown in vivo. We will confirm ability to promote regeneration using an optic nerve model because retinal ganglion cells are known to respond to PTEN knockdown. Safety will be assessed by following clinical signs and clinical chemistry.

项目摘要针对多个且经过验证的治疗靶标的创新药物/治疗组合具有在SCI后大大改善预后的潜力。关于轴突再生和啮齿动物的恢复表明，脊髓不是硬有线，并且可能发生“学习” 在脊髓电路中。引起轴突再生和发芽的化合物有助于可塑性至少在啮齿动物中，脊髓和大幅度改善了创伤性损伤的恢复。在啮齿动物中测试的许多不同的化合物/疗法会促进再生。几乎没有例外，促进再生的化合物还可以改善脊柱的功能恢复绳索受伤。最近，从缺失的删除中获得了非常令人印象深刻的轴突再生 PTEN。该应用程序的目的是同时评估PTEN作为治疗目标脊髓损伤的治疗和一种新型的RNAi递送技术可提供长时间的体内基因敲低效应。我们将创建“自我交付”小干扰RNA（SDRNA）针对PTEN。引入SDRNA分子的化学修饰使它们成为细胞透水。已证明该技术可以在几种应用中在体内工作，包括交货到视网膜。作为第一步，我们将合成一组SDRNA序列并在哺乳动物细胞培养物中评估它们以选择PTEN的铅候选者击倒。接下来，我们将测试神经元细胞培养物中的化合物，然后使用主要神经元。我们还将研究对原发性神经胶质的形态和增殖的影响细胞和人胶质母细胞瘤细胞。一旦我们确定了促进的主要候选人稳健的神经突增长，我们将测试化合物在体内敲低的功效。我们将确认使用视神经模型促进再生的能力，因为视网膜神经节已知细胞对PTEN敲低反应。安全将通过以下临床评估体征和临床化学。