Gene therapy for tuberous sclerosis

结节性硬化症的基因治疗

基本信息

批准号：
9810206
负责人：
XANDRA OWENS BREAKEFIELD
金额：
$ 42万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9810206
关键词：
Affect Alleles Animals Behavior Binding Biochemical Biochemistry Biological Assay Biotechnology Birth Brain C-terminal CCI-779 Caring Cell Proliferation Cell Size Cells Cellular biology Cerebral Ventricles Cessation of life Child Clinical Clinical Trials Complication Craniotomy Dependovirus Development Disease Dose Effectiveness Embryo Ensure Enzymes Epilepsy Evaluation FRAP1 gene Fibroblasts GTPase-Activating Proteins Genome Genotype Growth Factor Hamartoma Histopathology Human Hydrocephalus Immunohistochemistry Impaired cognition Impaired health Inherited Injections Institution Intellectual Property Intravenous Kidney Knockout Mice Length Lesion Licensing Life Liver Longevity LoxP-flanked allele Lung Magnetic Resonance Imaging Measurement Mental Health Methyl-CpG-Binding Protein 2 Modeling Monitor Morbidity - disease rate Mus Mutation N-terminal Neuraxis Neuroanatomy Neurologic Neurons Nodule Organ Pathology Patients Peripheral Phase Physiological Procedures Property Property Rights Proteins Resistance Rodent Secure Seizures Serotyping Serum Signal Transduction Sirolimus Subependymal Subependymal Giant Cell Astrocytoma Symptoms TSC1 gene TSC2 gene Testing Time Tissues Toxic effect Treatment Efficacy Tuberous Sclerosis Tuberous sclerosis protein complex Tumor Suppressor Proteins Ursidae Family Validation Ventricular Work adeno-associated viral vector autism spectrum disorder c-myc Genes cell type clinical care clinically relevant compare effectiveness dosage experimental study gene replacement gene therapy induced pluripotent stem cell loss of function mouse model nerve stem cell nervous system disorder neuropathology novel overexpression pre-clinical pre-clinical research prevent promoter response vector virus Cre recombinase

项目摘要

Our team will evaluate the potential for gene therapy to reduce life threatening symptoms in tuberous sclerosis complex (TSC), which affects 2 million people world-wide. TSC is an autosomal dominant, tumor suppressor disorder caused by an inherited mutation in either TSC1 (encoding hamartin) or TSC2 (encoding tuberin) with somatic loss of the corresponding normal allele leading to hamartomas (focal cell overgrowths) in different organs. Neurological involvement includes cortical tubers, subependymal nodules and subependymal giant cell astrocytomas with hydrocephalus epilepsy, autism, cognitive impairment and mental health issues. We have created a stochastic central nervous system (CNS) mouse model of TSC2 which manifests subependymal overgrowths, hydrocephalus and early death, reflecting what happens in TSC patients. Current standards-of- care for subependymal lesions in the brain involve neurosurgical craniotomy and/or long term treatment with rapamycin, both having potentially damaging effects on brain development. Our thesis is that a life-threatening complication of TSC – hydrocephalus can be prevented with a less invasive and longer lasting procedure, i.e. intravenous (IV) delivery of an adeno-associated virus (AAV) vector encoding a replacement protein, without compromising use of other standards-of-care if needed. IV delivery should achieve “extra copies” of the replacement gene in peripheral tissues reducing the likelihood of other life threatening hamartomas forming in the body. We will use our CNS Tsc2 mouse model to evaluate the ability of an AAV vector encoding a condensed version of tuberin (cTuberin) to reduce the size of ependymal abnormalities and extend lifespan. R61 Aim 1 - Evaluation of cTuberin protein in culture, including use of induced pluripotent stem cells from patients differentiated into neural progenitor cells, with cytoxicity and functional assays, Go-no-go: cTuberin must show at least 50% of tuberin activity in suppression of mTOR activity, and normalization of cell size and rate of cell proliferation. Aim 2 - Optimize AAV serotype and promoter with AAV-cTuberin in CNS Tsc2 mouse model and evaluate therapeutic efficacy. Behavior, survival times, neuropathology and whole body pathology will be monitored, as well as vector distribution and the ability of the vector to target cells in the brain and peripheral tissues. Go-no-go: AAV-cTuberin must at least double the average lifespan in this CNS Tsc2 model and show comparable effectiveness to rapamycin (positive control). In R33 Aim 3 - Evaluate dose escalation and potential toxicity of AAV-cTuberin in Tsc2+/- and CNS Tsc2 mouse models, as well as obtaining quantitative measurements of ventricular volumes in treated and non-treated mice by magnetic resonance imaging. Our team of TSC experts includes Drs. Breakefield (preclinical gene therapy for neurologic diseases), Maguire (AAV vectors), Ramesh (biochemistry and cell biology), Stemmer-Rachamimov (neuroanatomy) and Thiele (clinical care). These activities are aimed to carry out preclinical research in support of clinical trials, to secure intellectual property rights through our institution, and to facilitate licensing to biotechnology companies.

我们的团队将评估基因治疗减少结节硬化症症状的潜力复杂（TSC），影响全球200万人。 TSC是常染色体显性肿瘤抑制剂由TSC1（编码Hamartin）或TSC2（编码Tuberin）中的遗传突变引起的障碍相应的正常等位基因的体细胞丧失导致不同的Hamartomas（局灶性细胞过度生长）器官。神经学的参与包括皮质块茎，依赖型结节和依赖型巨细胞与脑积水癫痫，自闭症，认知障碍和心理健康问题的星形胶质细胞瘤。我们有创建了TSC2的随机中枢神经系统（CNS）小鼠模型，该模型表现过度生长，脑积水和早期死亡，反映了TSC患者发生的情况。当前标准护理大脑中的亚依赖膜病变涉及神经外科颅骨切开术和/或长期治疗雷帕霉素都对脑发育有潜在的破坏作用。我们的论点是威胁生命 TSC - 脑积水的并发症可以通过侵入性较小且持久的过程（即静脉注射（IV）递送腺相关病毒（AAV）载体编码替代蛋白，而无需如果需要，会妥协使用其他标准。静脉输送应获得的“额外副本” 外周组织中的替换基因降低了其他威胁生命的哈马瘤的可能性身体。我们将使用CNS TSC2鼠标模型来评估编码AAV矢量的能力凝结的Tuberin（CTUBERIN），以减少室系异常的大小并延长寿命。 R61 AIM 1-培养中ctuberin蛋白的评估，包括使用诱导的多能干细胞患者分化为具有细胞毒性和功能测定的神经祖细胞，go-no-go：ctuberin 必须在MTOR活性抑制以及细胞大小的归一化和细胞增殖速率。 AIM 2-在CNS TSC2鼠标中使用AAV -CTUBERIN优化AAV血清型和启动子建模并评估治疗效率。行为，生存时间，神经病理学和全身病理学将受到监控，以及向量分布以及矢量靶向大脑中细胞的能力，外围组织。 GO-NO-GO：AAV-CTUBERIN必须至少是此CNS TSC2模型中平均寿命的两倍并显示出与雷帕霉素的可比性（阳性对照）。在R33 AIM 3中 - 评估剂量升级 AAV-CTUBERIN在TSC2 +/-和CNS TSC2小鼠模型中的潜在毒性，并获得通过磁共振对处理和未处理的小鼠中心室体积的定量测量成像。我们的TSC专家团队包括DRS。 Breakefield（神经系统疾病的临床前基因治疗）， Maguire（AAV载体），Ramesh（生物化学和细胞生物学），Stemmer-Rachamimov（神经解剖学）和 Thiele（临床护理）。这些活动的目的是进行临床前研究，以支持临床试验，以通过我们的机构确保实质性财产权，并促进向生物技术公司许可。