Optogenetic and Chemogenetic Dissection of Cell Transplants

细胞移植的光遗传学和化学遗传学解剖

基本信息

批准号：
10617567
负责人：
THYAGARAJAN SUBRAMANIAN
金额：
$ 36.12万
依托单位：
UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
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：
2022-05-04 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10617567
关键词：
Agonist Allografting American Antiparkinson Agents Axon Back Basal Ganglia Behavior Behavior assessment Behavioral Biochemical Cell Line Cell Nucleus Cell Transplantation Cells Chlorides Clinical Clinical Trials Clozapine Corpus striatum structure Data Deep Brain Stimulation Disease Dissection Dopamine Dopamine Receptor Dopaminergic Cell Dose Drug-Induced Dyskinesia Electroencephalography Electrophysiology (science)Ensure Europe Exhibits Female Functional disorder Future GTP-Binding Proteins Genes Grant Halorhodopsins High Pressure Liquid Chromatography High Prevalence Histology Human Implant Inflammatory Response Internal Ribosome Entry Site Lasers Lesion Letters Levodopa Lifting Light LoxP-flanked allele Mediating Methods Microdialysis Microelectrodes Modeling Monkeys Morbidity - disease rate Motor Motor Cortex Muscarinic Acetylcholine Receptor Neurons Operative Surgical Procedures Opsin Oxidopamine Parkinson Disease Parkinsonian Disorders Patients Pattern Phototoxicity Proteins Rattus Regulation Reproducibility Research Resolution Role Safety Substantia nigra structure Symptoms Synapses Synaptic Receptors System Techniques Testing Therapeutic Time Transplantation United States National Institutes of Health awake behavioral study clinical investigation clinical translation connectome designer receptors exclusively activated by designer drugs disability dopaminergic neuron experimental study fetal graft function histological studies improved in vivo inclusion criteria male medical complication neurochemistry neurophysiology novel therapeutics optogenetics parkinsonian rodent pars compacta receptor recombinant viral vector reuptake side effect stem cells vector

Agonist Allografting American Antiparkinson Agents Axon Back Basal Ganglia Behavior Behavior assessment Behavioral Biochemical Cell Line Cell Nucleus Cell Transplantation Cells Chlorides Clinical Clinical Trials Clozapine Corpus striatum structure Data Deep Brain Stimulation Disease Dissection Dopamine Dopamine Receptor Dopaminergic Cell Dose Drug-Induced Dyskinesia Electroencephalography Electrophysiology (science)Ensure Europe Exhibits Female Functional disorder Future GTP-Binding Proteins Genes Grant Halorhodopsins High Pressure Liquid Chromatography High Prevalence Histology Human Implant Inflammatory Response Internal Ribosome Entry Site Lasers Lesion Letters Levodopa Lifting Light LoxP-flanked allele Mediating Methods Microdialysis Microelectrodes Modeling Monkeys Morbidity - disease rate Motor Motor Cortex Muscarinic Acetylcholine Receptor Neurons Operative Surgical Procedures Opsin Oxidopamine Parkinson Disease Parkinsonian Disorders Patients Pattern Phototoxicity Proteins Rattus Regulation Reproducibility Research Resolution Role Safety Substantia nigra structure Symptoms Synapses Synaptic Receptors System Techniques Testing Therapeutic Time Transplantation United States National Institutes of Health awake behavioral study clinical investigation clinical translation connectome designer receptors exclusively activated by designer drugs disability dopaminergic neuron experimental study fetal graft function histological studies improved in vivo inclusion criteria male medical complication neurochemistry neurophysiology novel therapeutics optogenetics parkinsonian rodent pars compacta receptor recombinant viral vector reuptake side effect stem cells vector

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项目摘要

Parkinson's disease (PD) causes disability in an estimated 1 million Americans and is characterized by progressive loss of dopaminergic neurons that originate in the substantia nigra pars compacta (SNpc) and their axons. Levodopa (LD) in some form is required to control symptoms of PD in vast majority of patients, but its use causes drug-induced dyskinesias (DID), motor and non motor fluctuations, disabling complications that remains a major problem in contemporary management of PD. Intermittent high dose LD treatments, loss of continuous dopaminergic stimulation (CDS) along with dopamine receptor super sensitivity are hypothesized as causative factors for DID and fluctuations. It is also well accepted that information transfer from the motor cortex to the basal ganglia and back is abnormal in PD and correction of electrophysiological abnormalities may be critical to provide symptomatic relief to patients with advanced PD. Our studies show that the electrophysiological changes that occur in the basal ganglia as result of parkinsonism remain largely unmitigated with round the clock LD treatments despite excellent amelioration of parkinsonian behavior. In contrast, dopaminergic cell transplants into the striatum “normalize” basal ganglia neurophysiology and parkinsonian behavioral abnormalities. We show that synaptic connectivity between the graft and the host is critical and necessary to mediate these effects. These findings suggest that pre-synaptic factors such as the presence of nigrostriatal synapses that enables focused release of dopamine, its reuptake and regulation via pre-synaptic receptors may all be important to minimize the motor complications of PD. Recent studies have shown that dopaminergic cell transplantation for PD can be successful and without deleterious side effects providing >2 decades of effective anti-parkinsonian benefits to patients. This has led to the lifting of the moratorium on clinical cell transplantation for PD. However, optimization of cell transplantation techniques including the ability to externally control the functioning of the grafts and its newly formed synaptic connections with the host is critical to ensure safety of future clinical translation of such grafts. Therefore, we seek to comprehensively evaluate the hypothesis that electrophysiological “normalization” in the basal ganglia – cortical circuit is necessary to mitigate or eliminate DID and fluctuations in PD. Using a highly reproducible rat model of PD that exhibit DID and fluctuations, optogenetic and chemogenetic modulation of grafts such that they can be turned “off” and “on”, awake basal ganglia single cell recordings, LFP recordings, EEG and in vivo microdialysis combined with HPLC-Ms/Ms we plan to study the biochemical and electrical abnormalities in the basal ganglia and the cortex associated with DID and fluctuations in PD. Multiple nuclei in the basal ganglia and its known connections will be probed. Extensive histological studies will also be performed. This research will provide mechanistic basis for the electrophysiological abnormalities that accompany DID, motor and non- motor fluctuations in PD and help discover methods to correct them.

帕金森氏病（PD）导致估计有100万美国人的残疾，其特征是逐渐丧失起源于黑质的nigra pars commacta（SNPC）及其其多巴胺能神经元及其轴突。左旋多巴（LD）以某种形式需要控制绝大多数患者的PD症状，但它使用引起药物引起的运动障碍（DID），运动和非运动波动，使并发症破坏并发症仍然是PD当代管理的主要问题。间歇性高剂量LD治疗，丧失假设连续多巴胺能刺激（CD）以及多巴胺受体超级灵敏度作为DID和波动的原因。信息从电动机传输也很广泛基底神经节和背部的皮质在PD中异常和电生理异常的校正对于晚期PD患者提供症状缓解可能至关重要。我们的研究表明由于帕金森氏症的结果，基底神经节发生的电生理变化在很大程度上仍然存在圆形LD治疗不受限制地渴望对帕金森氏症行为的出色改善。在对比，多巴胺能细胞移植到纹状体“正常化”基底神经神经生理学和帕金森氏行为异常。我们表明，移植物与宿主之间的合成连通性是至关重要的，需要调解这些效果。这些发现表明，突触前因素，例如存在质纹状体突触的存在，可以重点释放多巴胺，其再摄取和调节突触前受体对于最大程度地减少PD的运动并发症可能很重要。最近的研究表明PD的多巴胺能细胞移植可以成功，并且没有有害的副作用为患者提供> 20年有效的反 - 公园福利。这导致了暂停PD临床细胞移植。但是，优化细胞移植技术包括外部控制移植物的功能及其新形成的突触连接的能力宿主对确保此类移植物的未来临床翻译的安全至关重要。因此，我们寻求全面评估了基底神经节中电生理“归一化”的假设 - 减轻或消除DID并在PD中波动是必要的。使用高度可再现的老鼠暴露的PD模型以及移植的波动，光学遗传学和化学遗传调节的模型它们可以“关闭”和“ ON”，清醒的巴萨神经节录音，LFP录音，脑电图和体内微透析结合HPLC-MS/MS，我们计划研究生化和电气异常基底神经节和与DID相关的皮质以及PD中的波动。基底神经节中的多个核并将探测其已知的连接。还将进行广泛的组织学研究。这项研究将为可容纳的电生理异常提供机械基础，运动和非 - PD中的电动机波动，并有助于发现纠正它们的方法。