Choroid Plexus-Directed Gene Therapy for Alpha-Mannosidosis

脉络丛定向基因治疗α-甘露糖苷贮积症

• 批准号: 9203064
• 负责人: STEPHEN GERARD KALER
• 金额: $ 53.02万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-24 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9203064
• 关键词: Affect; Alpha-mannosidase; Animal Model; Animals; Award; Biochemical; Biological Markers; Biological Products; Blood; Blood Vessels; Brain; Brain Diseases; Brain Pathology; Cells; Cerebral Ventricles; Cerebrospinal Fluid; Cerebrum; Choroid Plexus Epithelium; Clinical; Clinical Management; Clinical Trials; Collection; Complementary DNA; Copper; Data; Dependovirus; Development; Disease; Dose; Drug Approval; Enzymes; Epithelial Cells; Evaluation; Extracellular Space; Extramural Activities; Face; Felis catus; Future; Gene Transfer; Gene therapy trial; Genes; Goals; Grant; Health; Histologic; Human; Hybrids; Immune; Inborn Genetic Diseases; Infusion procedures; Injectable; Intellectual functioning disability; Intrathecal Injections; Intraventricular Injections; Investigational Drugs; Investigational New Drug Application; Ions; Kale - dietary; Laboratories; Lesion; Link; Lysosomal Storage Diseases; Magnetic Resonance; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mediating; Menkes Kinky Hair Syndrome; Methods; Modeling; Mus; Natural History; Neurologic; Outcome Measure; Pathology; Patient-Focused Outcomes; Patients; Penetration; Pharmacology and Toxicology; Phase; Phase I Clinical Trials; Phenotype; Pilot Projects; Process; Production; Proteins; Protocols documentation; Recombinant adeno-associated virus (rAAV); Recombinants; Research Design; Research Personnel; Resolution; Safety; Series; Serotyping; Source; Spinal Tap; Structure; Structure of choroid plexus; Subarachnoid Space; Surrogate Markers; Testing; Therapeutic Effect; Toxic effect; Toxicology; Translating; United States National Institutes of Health; Water; Work; adeno-associated viral vector; alpha-Mannosidosis; base; bench to bedside; biomarker identification; cerebrospinal fluid flow; cisterna magna; clinical biomarkers; clinical practice; enzyme replacement therapy; experimental study; gene therapy; hearing impairment; human subject; innovation; insight; interest; lateral ventricle; man; mouse model; nonhuman primate; novel; phase I trial; pre-clinical; preclinical development; preclinical study; public health relevance; risk benefit ratio; skeletal abnormality; spectroscopic imaging; transgene expression; treatment effect; vector; ventricular system

 DESCRIPTION (provided by applicant): This proposal links a series of innovative AAV gene therapy experiments in two well-established animal models of alpha-mannosidosis with preclinical toxicology studies to develop a path forward to a first-in-human clinical trial of cerebrospinal fluid (CSF)-delivered gene therapy for this illness. The Kaler lab has treated a mouse model of Menkes disease using CSF-directed AAV5 gene therapy via efficient transduction of choroid plexus epithelia. The choroid plexuses (CP) are highly vascularized structures that project into the ventricles of the brain. The polarized epithelia of the CP produce CSF and secrete a large number of proteins. Lysosomal storage diseases (LSDs) could potentially benefit from a CP-targeted approach because AAV transduction results in sustained episomal transgene expression and CP epithelia have a slow rate of turnover. The Wolfe lab has shown that AAV gene transfer into the brains of animal models of LSDs can mediate substantial but incomplete cellular correction and clinical improvement. The advantage of targeting CSF flow is that it extends throughout the ventricular system to the subarachnoid space, from which molecules ultimately reach the entire brain. Intrathecal delivery of purified recombinant lysosomal enzyme (enzyme replacement therapy, ERT) has been successful in ameliorating some brain pathology in animal models and recently in a human clinical trial. However, a major drawback to this approach is the need for repeated intrathecal injections due to short half- lives of recombinant enzymes. An alternative long-term strategy is to remodel CP epithelial cells with an AAV vector to secrete enzyme of interest. CP epithelia have an extremely slow turnover rate, thus this approach could generate a permanent source of enzyme production for secretion into the CSF and penetration to brain structures. Furthermore, intraventricular injection AAV vectors can deliver the normal gene to some of the parenchymal cells, which can then also secrete normal enzyme into extracellular spaces for delivery to other cells. A two-year NIH Bench-to-Bedside Award (R21 equivalent) supported the initial evaluation of this hypothesis in animal models of alpha-mannosidosis (AMD), a prototypical LSD. This U01 proposal expands that effort with the goal of completing the pre-clinical studies needed to submit a phase I clinical trial IND in 3-4 years. We propose to: 1) Evaluate the relative abilities of different AAV-vector serotypes to support CSF- directed AAV transduction in a mouse model of AMD and test the most promising serotypes in the much larger brain of the AMD cat, with the goal of identifying the vector that is most likely to produce the greatest benefit in human clinica trials; 2) Establish clinical and biochemical features in AMD patients for outcome measures in a future clinical trial; and 3) Perform preclinical toxicology studies in non-human primates and develop regulatory approval for a first-in-human gene therapy trial for AMD. The potential impact on clinical practice in the field of LSD is high since, if the proposed aims are successfully achieved, the largest current barriers to health for patients with AMD and other LSDs with brain disease would be circumvented.

 描述（由申请人提供）：该提案将在两个完善的α-甘露糖苷沉积症动物模型中进行的一系列创新AAV基因治疗实验与临床前毒理学研究联系起来，以开发一条通往针对这种疾病的脑脊液（CSF）递送基因治疗的首次人体临床试验的道路。Kaler实验室通过有效转导脉络丛上皮细胞，使用CSF导向的AAV 5基因治疗治疗Menkes病小鼠模型。脉络丛（CP）是高度血管化的结构，突出到脑室。CP的极化上皮细胞产生 CSF并分泌大量蛋白质。溶酶体贮积病（LSD）可能受益于CP靶向方法，因为AAV转导导致持续的附加型转基因表达，并且CP上皮细胞具有缓慢的更新速率。Wolfe实验室已经证明，将AAV基因转移到LSD动物模型的大脑中可以介导大量但不完全的细胞校正和临床改善。靶向CSF流动的优点是它延伸到整个脑室系统到蛛网膜下腔，分子最终从蛛网膜下腔到达整个大脑。鞘内递送纯化的重组溶酶体酶（酶替代疗法，ERT）已经成功地改善动物模型中的一些脑病理学，并且最近在人类临床试验中。然而，该方法的主要缺点是由于重组酶的短半衰期而需要重复鞘内注射。另一种长期策略是用AAV载体重塑CP上皮细胞以分泌感兴趣的酶。CP上皮细胞具有极低的周转率，因此这种方法可以产生用于分泌到CSF中并渗透到脑结构的酶产生的永久来源。此外，脑室内注射AAV载体可以将正常基因递送到一些实质细胞，其然后也可以将正常酶分泌到细胞外空间以递送到其他细胞。一项为期两年的NIH实验室到床边奖（相当于R21）支持在α-甘露糖苷酶（AMD）（一种原型LSD）动物模型中对这一假设的初步评估。U 01提案扩大了这一努力，目标是在3-4年内完成提交I期临床试验IND所需的临床前研究。我们建议：1）评估相对能力 2）在AMD患者中建立临床和生物化学特征，用于未来临床试验中的结果测量;和3）在非人灵长类动物中进行临床前毒理学研究，并为首次用于AMD的人类基因治疗试验开发监管批准。对LSD领域临床实践的潜在影响很大，因为如果成功实现拟议目标，将绕过AMD患者和其他患有脑部疾病的LSD患者目前最大的健康障碍。