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

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

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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.

项目摘要三分之二的溶酶体储存障碍（LSD）会影响大脑，但大多数LSD治疗都不会改善中枢神经系统（CNS）符号。大脑指导基因疗法的几项试验未能显示临床益处在中枢神经系统中恢复蛋白质表达。结果在主题中特别差已经开发了神经系统缺陷，表明单独拯救表达可能不足以纠正功能一旦建立了神经元电路。在CLN3疾病中，代表性的LSD和最常见的小儿痴呆症原因，患者发展失明，癫痫发作和痴呆症。已经开发了几种CLN3疾病小鼠模型。而全部概括患者中看到的存储积累，行为表型微妙而可变。到克服了这一限制，我们确定了网络级电生理学上的鲁棒，可再现的表型在两个CLN3模型中的研究，一个敲除和人类突变模型。与组织病理学不同，生理学措施直接反映了功能，因此可能是治疗发展的更好读数。我们的工作表明CLN3疾病传统上认为是退化性疾病，早期破坏了神经发育，特别是在海马，这是CLN3疾病中的脆弱区域。在体外电压上敏感的染料成像（VSDI）和体内脑电图（EEG）记录，CLN3 - / - 鼠标已改善海马齿状回（DG）的兴奋性，更快的EEG背景活动和海马丧失锋利的波浪纹波，编码新记忆的振荡。另外，DG神经发生也可能是更新的一种补偿机制，早期（2mo），但不晚于疾病（6MO）。其他网络变化也会发生神经变性的模型，包括阿尔茨海默氏病（AD）。深脑刺激内嗅已显示皮层可以改善AD小鼠模型中的结果。以前，我们发现P0处的很早的CLN3基因更换会纠正CLN3中的网络动力学淘汰鼠标。我们的中心假设是，异常的神经元电路发育将限制时间窗口，即“治疗窗口”，当基因置换将改善网络生理时 CLN3病。此外，我们预测，钥匙电路中的活动改变可能会改变治疗基因治疗的窗户和效率。我们的具体目的是：1）定义异常的牙科发育在CLN3疾病小鼠中，2）确定通过校正海马电路动力学的治疗窗口基因置换和3）测试如果修改内嗅皮层活性会改变电路缺陷和对基因的反应治疗。通过这种方式，我们将使用CLN3疾病作为代表性LSD，以探索网络活动和对基因治疗的反应。我们的长期目标是开发以网络为导向的疗法当与基因替代结合使用时，可以改善LSD的结果。