Characterizing and testing the efficacy of AAV-mediated gene therapy in a sheep model of CLN1 disease.

在 CLN1 疾病绵羊模型中表征和测试 AAV 介导的基因治疗的功效。

基本信息

批准号：
10339842
负责人：
JONATHAN D COOPER
金额：
$ 55.06万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10339842
关键词：
Address Affect Animals Biodistribution Brain Brain region CLN1 gene CLN2 gene CRISPR/Cas technology Canis familiaris Characteristics Child Childhood Clinic Clinical Clinical Treatment Clinical Trials Cognitive Complex Data Development Disease Disease Progression Disease model Dose Enzymes FDA approved Gene Delivery Goals Histologic Human Image Immune response Infantile neuronal ceroid lipofuscinosis Inherited Injections Knowledge Life Longevity Magnetic Resonance Imaging Mediating Mus Nerve Degeneration Nervous System Physiology Neuraxis Neurodegenerative Disorders Neurologic Neuronal Ceroid-Lipofuscinosis Organ Outcome Outcome Measure Pathogenesis Pathology Performance Phenotype Positioning Attribute Program Development Publishing Radiology Specialty Regimen Route Safety Serotyping Sheep Site Spielmeyer-Vogt Disease Spinal Cord Testing Therapeutic Therapeutic Effect Time Translating Treatment Efficacy Treatment outcome Ursidae Family Viral Viral Genes Visceral Work adeno-associated viral vector base brain magnetic resonance imaging canine model cerebral atrophy clinical translation clinically translatable cognitive function delivery vehicle design disease phenotype disease-causing mutation early onset effective therapy efficacy testing end stage disease enzyme activity experience gene therapy genome editing human model improved in vivo infancy mouse model nervous system disorder novel research clinical testing response scale up sheep model therapeutic evaluation therapy outcome thioesterase PPT1 gene product vector

项目摘要

Project Summary/Abstract: CLN1 disease or Infantile Neuronal Ceroid Lipofuscinosis (INCL or Infantile Batten disease) is one of the earliest onset and most rapidly progressing forms of neuronal ceroid lipofuscinosis (NCL or Batten disease). CLN1 disease is caused by deficiency in the lysosomal enzyme palmitoyl protein thioesterase-1 (PPT1). This deficiency has a devastating and rapidly progressing effect upon affected children that starts within the first year of life, and because there is no effective therapy available CLN1 disease is always fatal. We have been able to dramatically improve therapeutic outcomes in PPT1-deficient mice by targeting adeno-associated viral (AAV)-mediated gene therapy to the central nervous system (CNS) regions that are most affected, including the spinal cord. However, unlike other forms of NCL, such therapeutic effects are limited to the CNS regions that are transduced. As such, successfully translating gene therapy for CLN1 disease into the clinic will be a significant challenge in the much larger and more complex brain of a child. To overcome this obstacle, we shall use a novel CRISPR/Cas9 generated CLN1 R151X sheep model to refine our therapeutic strategy, assessing the delivery, dosing, safety and efficacy of gene therapy in a larger species that is ideally suited for translating these advances. We recently published that these CLN1 R151X sheep display pronounced CLN1 disease-relevant phenotypes. Our preliminary data extend these observations, revealing an earlier onset of neurologic disease, and widespread histologically and radiologically detectable pathology that is more pronounced than in PPT1-deficient mice. We have also shown that an intracranial injection of an AAV9 vector expressing PPT1 raises expression of this enzyme in the brain of sheep to supraphysiological levels. We believe CLN1 R151X sheep not only more accurately model human CLN1 disease, but also provide an ideal testing ground for optimizing the dosing and delivery of gene therapy in a fashion that is not possible in mice. We now propose to characterize the progression of these disease phenotypes in CLN1 R151X sheep, in order to provide detailed landmarks of disease progression using cognitive and neurologic testing, and MRI imaging, correlating these data with histological findings (Aim 1). We will also define the parameters of vector dosing and delivery routes to achieve widespread transduction of the sheep brain and spinal cord, and elevate PPT1 activity to levels predicted to be capable of producing therapeutic benefit (Aim 2). Finally, we shall determine the therapeutic efficacy, minimum effective dose, safety and clinical response to this optimized delivery of scAAV9-CCAG-PPT1 to the brain and spinal cord of CLN1 R151X sheep (Aim 3). These data will allow us to refine our gene therapy approach, and position us for entry into the CREATE-Bio development program towards clinical translation of the first effective treatment of this devastating disease.

项目摘要/摘要： CLN1 病或婴儿神经元蜡质脂褐质沉着症（INCL 或婴儿 Batten 病）是其中一种神经元蜡质脂褐素沉积症（NCL 或 Batten 病）最早发病且进展最快的形式。 CLN1 疾病是由溶酶体酶棕榈酰蛋白硫酯酶-1 (PPT1) 缺陷引起的。这缺乏症对受影响的儿童产生毁灭性的、迅速进展的影响，这种影响从一开始就开始出现。由于没有有效的治疗方法，CLN1 疾病总是致命的。我们有通过靶向腺相关蛋白，能够显着改善 PPT1 缺陷小鼠的治疗结果病毒（AAV）介导的针对受影响最严重的中枢神经系统（CNS）区域的基因治疗，包括脊髓。然而，与其他形式的 NCL 不同，这种治疗效果仅限于中枢神经系统被转导的区域。因此，成功地将 CLN1 疾病的基因疗法转化为临床将对于孩子更大、更复杂的大脑来说，这是一个重大挑战。为了克服这个障碍，我们应使用新型 CRISPR/Cas9 生成的 CLN1 R151X 绵羊模型来完善我们的治疗策略，评估基因治疗在较大物种中的传递、剂量、安全性和有效性，这是理想的适合转化这些进步。我们最近发布了这些 CLN1 R151X 羊展示明显的 CLN1 疾病相关表型。我们的初步数据扩展了这些观察结果，揭示了神经系统疾病发病较早，组织学和放射学上可检测到广泛的病理学，比 PPT1 缺陷小鼠更明显。我们还表明，颅内注射 AAV9 表达 PPT1 的载体将绵羊大脑中这种酶的表达提高到超生理水平。我们相信 CLN1 R151X 羊不仅可以更准确地模拟人类 CLN1 疾病，而且还提供了以一种不可能的方式优化基因治疗的剂量和递送的理想试验场老鼠。我们现在建议描述 CLN1 R151X 绵羊中这些疾病表型的进展特征，为了使用认知和神经学测试以及 MRI 提供疾病进展的详细标志成像，将这些数据与组织学结果相关联（目标 1）。我们还将定义向量的参数剂量和递送途径以实现绵羊大脑和脊髓的广泛转导，并提高 PPT1 活性达到预计能够产生治疗效果的水平（目标 2）。最后，我们将确定这种优化的治疗效果、最小有效剂量、安全性和临床反应将 scAAV9-CCAG-PPT1 递送至 CLN1 R151X 绵羊的大脑和脊髓（目标 3）。这些数据将让我们能够完善我们的基因治疗方法，并为我们进入 CREATE-Bio 开发做好准备旨在将这种毁灭性疾病的第一个有效治疗方法临床转化的计划。