Network modulation to improve gene therapy in CLN3 disease

网络调节改善 CLN3 疾病的基因治疗

基本信息

批准号：
10626675
负责人：
Rebecca Clare Ahrens-Nicklas
金额：
$ 47.18万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
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 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10626675
关键词：
Address Adolescent Affect Age Alzheimer&apos s Disease Alzheimer&apos s disease model Blindness Brain CLN3 gene Cerebrum Cessation of life Child Childhood Chronic Clinical Clinical Trials Data Deep Brain Stimulation Defect Degenerative Disorder Dementia Development Diagnosis Disease Dose Electroencephalogram Electrophysiology (science)Fimbria of hippocampus Functional disorder Gene Expression Glutamates Goals Hippocampus (Brain)Histology Histopathology Housing Human Image In Vitro Knock-out Knockout Mice Measures Memory Modeling Mus Mutation Nerve Degeneration Neuraxis Neurologic Deficit Neurologic Symptoms Neuronal Ceroid-Lipofuscinosis Neurons Outcome Pathologic Pathology Patients Phenotype Physiological Physiology Process Progressive Disease Reporting Reproducibility Seizures Symptoms Testing Therapeutic Time Treatment Efficacy Variant Work base behavioral phenotyping clinical practice dentate gyrus designer receptors exclusively activated by designer drugs entorhinal cortex enzyme replacement therapy functional restoration gene replacement gene therapy improved improved outcome in utero in vivo mouse model neurodevelopment neurogenesis neuronal circuitry novel strategies protein expression response therapy development voltage sensitive dye

项目摘要

PROJECT SUMMARY Two-thirds of lysosomal storage disorders (LSD) affect the brain, yet most LSD treatments do not improve central nervous system (CNS) symptoms. Several trials of brain-directed gene therapy have failed to show clinical benefit despite restoring protein expression in the CNS. Outcomes are especially poor in subjects who have developed neurologic deficits, suggesting rescue of expression alone may be insufficient to correct function once diseased neuronal circuits are established. In CLN3 Disease, a representative LSD and the most common cause of pediatric dementia, patients develop blindness, seizures, and dementia. Several CLN3 disease mouse models have been developed. While all recapitulate the storage accumulation seen in patients, behavioral phenotypes are subtle and variable. To overcome this limitation, we identified robust, reproducible phenotypes on network-level electrophysiology studies in two CLN3 models, a knockout and a human mutation model. Unlike histopathology, physiologic measures directly reflect function and, therefore, may be a better readout for therapy development. Our work suggests CLN3 disease, traditionally considered a degenerative disorder, disrupts early neurodevelopment, especially in the hippocampus, a vulnerable region in CLN3 disease. On in vitro voltage sensitive dye imaging (VSDI) and in vivo electroencephalogram (EEG) recordings, Cln3-/- mice have decreased excitability of the hippocampal dentate gyrus (DG), faster EEG background activity, and loss of hippocampal sharp wave ripples, oscillations that encode new memories. Also, DG neurogenesis is upregulated, perhaps as a compensatory mechanism, early (2mo) but not later (6mo) in disease. Similar network changes arise in other models of neurodegeneration including Alzheimer’s disease (AD). Deep brain stimulation of the entorhinal cortex has been shown to improve outcomes in mouse models of AD. Previously, we found that very early Cln3 gene replacement at p0 corrects network dynamics in a Cln3 knockout mouse. Our central hypothesis is that abnormal neuronal circuit development will limit the window of time, i.e. ‘therapeutic window’, when gene replacement will improve network physiology in CLN3 disease. Furthermore, we predict that altering activity in a key circuit could modify the therapeutic window and efficacy of gene therapy. Our Specific Aims are to: 1) define abnormal dentate gyrus development in CLN3 disease mice, 2) determine the therapeutic window for correction of hippocampal circuit dynamics by gene replacement, and 3) test if modifying entorhinal cortex activity alters circuit defects and response to gene therapy. In this way we will use CLN3 Disease as a representative LSD, to explore the relationship between network activity and response to gene therapy. Our long-term goal is to develop network-directed therapies that, when combined with gene replacement, improve outcomes in LSDs.

项目概要三分之二的溶酶体贮积症 (LSD) 会影响大脑，但大多数 LSD 治疗并不能改善症状一些针对大脑的基因治疗试验未能显示出中枢神经系统（CNS）症状。尽管恢复了中枢神经系统中的蛋白质表达，但临床获益在受试者中尤其差。已经出现神经功能缺陷，仅挽救表达可能不足以纠正一旦患病的神经回路建立，就发挥功能。 CLN3 疾病是一种代表性的 LSD，也是儿童痴呆症的最常见原因，患者会出现目前已开发出几种 CLN3 疾病小鼠模型。概括一下患者体内的储存积累，行为表型是微妙且可变的。克服了这一限制，我们在网络级电生理学上发现了稳健的、可重复的表型与组织病理学、生理学不同，两个 CLN3 模型（敲除模型和人类突变模型）的研究。测量值直接反映功能，因此可能是治疗开发的更好读数。我们的工作表明，传统上被认为是一种退行性疾病的 CLN3 疾病会破坏早期神经发育，尤其是海马体，这是 CLN3 疾病中体外电压的脆弱区域。敏感染料成像（VSDI）和体内脑电图（EEG）记录，Cln3-/-小鼠的数量减少海马齿状回 (DG) 的兴奋性、更快的脑电图背景活动以及海马的丧失尖锐的波涟漪，编码新记忆的振荡，此外，DG 神经发生也被上调，也许是这样。一种补偿机制，在疾病的早期（2个月）而不是后期（6个月）也出现了类似的网络变化。神经退行性疾病模型，包括阿尔茨海默病（AD）的内嗅深部脑刺激。皮层已被证明可以改善 AD 小鼠模型的结果。之前，我们发现 p0 处的早期 Cln3 基因替换可以纠正 Cln3 中的网络动态我们的中心假设是异常的神经回路发育会限制时间窗口，即“治疗窗口”，此时基因替换将改善网络生理学此外，我们预测改变关键回路的活性可能会改变治疗效果。我们的具体目标是：1）定义齿状回发育异常。在 CLN3 疾病小鼠中，2) 通过以下方法确定纠正海马回路动力学的治疗窗口基因替换，3) 测试修改内嗅皮层活动是否会改变电路缺陷和对基因的反应这样我们就以CLN3疾病作为LSD的代表，来探讨两者之间的关系。我们的长期目标是开发网络导向疗法。与基因替代相结合，可以改善 LSD 的治疗结果。