Characterizing and testing the efficacy of AAV-mediated gene therapy in a novel CRISPR/Cas9 generated sheep model of Cln1 disease.

在新型 CRISPR/Cas9 生成的 Cln1 疾病绵羊模型中表征和测试 AAV 介导的基因治疗的功效。

• 批准号: 10357987
• 负责人: JONATHAN D COOPER
• 金额: $ 50.24万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10357987
• 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; 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; Methods; Modeling; Mus; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurologic; Neuronal Ceroid-Lipofuscinosis; Outcome; Outcome Measure; Pathogenesis; Pathology; 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; Work; adeno-associated viral vector; base; cerebral atrophy; clinical translation; clinically translatable; design; disease phenotype; disease-causing mutation; early onset; effective therapy; efficacy testing; end stage disease; enzyme activity; experience; gait examination; gene therapy; genome editing; human disease; human model; improved; in vivo; infancy; mouse model; mutant; nervous system disorder; novel; research clinical testing; response; scale up; sheep model; therapeutic evaluation; therapy outcome; thioesterase PPT1 gene product; vector

PROJECT SUMMARY 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 both MRI and MRS 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 CLN1疾病或婴儿神经元蛋白脂肪促脂肪促脂肪FIPOFOFOSISOS（含或婴儿the病）是其中之一 最早的发病和最快进展的神经元粘膜脂肪促脂肪促脂肪促脂肪促脂肪促脂（NCL或Batten疾病）。 CLN1疾病是由溶酶体葡萄酰蛋白硫代酶1（PPT1）缺乏引起的。这 缺乏症对受影响的儿童产生毁灭性和快速进步的影响 生命的一年，并且由于没有有效的疗法可用的CLN1疾病总是致命的。我们有 能够通过靶向腺相关，能够显着改善PPT1缺陷小鼠的治疗结果 病毒（AAV）介导的基因治疗对受影响最大的中枢神经系统（CNS）区域 包括脊髓。但是，与其他形式的NCL不同，这种治疗作用仅限于CNS 被转导的区域。因此，将CLN1疾病的基因治疗成功地转化为诊所 在孩子的大脑更大，更复杂的大脑中，要成为一个重大挑战。为了克服这一障碍，我们 应使用新颖的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分娩，剂量，安全性和功效 适合翻译这些进步。我们最近发表了这些CLN1 R151X绵羊展示 明显的与CLN1疾病相关的表型。我们的初步数据扩展了这些观察结果，揭示了 神经系统疾病的早期发作，并在组织学和放射学上广泛的病理学广泛发作 比PPT1缺陷型小鼠更为明显。我们还表明，颅内注入AAV9 表达PPT1的载体将这种酶在绵羊大脑中的表达提高到超生理水平。 我们认为CLN1 R151X绵羊不仅更准确地对人类CLN1疾病进行了建模，还提供了一种 理想的测试场，以优化基因疗法的剂量和递送的方式是不可能的 老鼠。现在，我们建议在Cln1 R151X绵羊中表征这些疾病表型的进展 为了使用MRI和MRS成像提供详细的疾病进展的地标，与之相关 这些数据带有组织学发现（AIM 1）。我们还将定义向量给予和交付的参数 实现绵羊大脑和脊髓的广泛转导的路线，并将PPT1活性提升到 预计能够产生治疗益处的水平（AIM 2）。最后，我们将确定 治疗功效，最低有效剂量，安全性和对这种优化的SCAAV9-的临床反应 CCAG-PPT1到CLN1 R151X绵羊的大脑和脊髓（AIM 3）。这些数据将使我们能够完善我们的 基因治疗方法，并将我们定位为进入创建生物开发计划 对这种毁灭性疾病的首次有效治疗的翻译。