Novel Re-engineered L DOPA Probiotic Therapy for Parkinson's Disease

新型重新设计的左旋多巴益生菌疗法治疗帕金森病

基本信息

批准号：
10688149
负责人：
Anumantha Gounder Kanthasamy
金额：
$ 36.47万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10688149
关键词：
Acute Address Adverse effects Anabolism Animal Model Area Bacteria Benserazide Bradykinesia Brain Carbidopa Chromosomal Stability Chromosomes Chromosomes, Human, 16-18 Chronic Clinical Codon Nucleotides Communities Complex Complication Corpus striatum structure Data Development Disease Dopamine Dopamine Receptor Dose Drug Kinetics Dyskinetic syndrome Engineered Probiotics Engineering Escherichia coli Exhibits Friends Generations Genes Genetic Genetic Engineering Goals Health Human Impaired cognition In Situ In Vitro Inbred C57BL Mice Iowa L-DOPA induced dyskinesia Legal patent Lesion Levodopa Measures Mixed Function Oxygenases Modality Modeling Modern Medicine Motor Mus Nature Nerve Degeneration Neurotoxins Oral Oral Administration Oxidopamine Oxidoreductase Parkinson Disease Parkinsonian Disorders Pathway interactions Patients Performance Pharmaceutical Preparations Pharmacodynamics Pharmacologic Substance Phase Plasma Plasmids Probiotics Production Quality of life Rhamnose Ribosomal RNA Rodent Model System Tablets Therapeutic Transgenic Organisms Treatment Efficacy Tremor Universities Work behavioral phenotyping blood-brain barrier crossing decarboxylase inhibitor delivery vehicle disease phenotype dopamine replacement therapy dopaminergic neuron gut colonization gut health gut microbiome host microbiota improved in vivo metagenomic sequencing microbial microbiome mitopark mouse motor deficit motor impairment motor symptom mouse model neurochemistry nigrostriatal pathway novel novel strategies novel therapeutics posture instability pre-clinical preclinical efficacy prevent probiotic supplementation probiotic therapy promoter reduce symptoms response side effect standard care stem success therapeutic development translational approach

项目摘要

Abstract The complex and prolonged disease course exhibited by Parkinson’s disease (PD) first starts with non- motor disturbances and then slowly progresses to mild-to-moderate motor deficits, ultimately inflicting severe movement impairment and cognitive decline. Dopamine deficiency resulting from nigrostriatal dopaminergic neuronal damage ultimately manifests as the cardinal extrapyramidal motor symptoms of rigidity, bradykinesia, tremors, and postural instability. This proposal addresses one of the greatest challenges facing the current anti- Parkinsonian therapy of dopamine replacement with the dopamine precursor L-DOPA. Currently, oral tablet dosing of L-DOPA/carbidopa 3-4 times/day remains the most effective and well-tolerated treatment, one that significantly improves the motor symptoms and quality of life of patients in the early stages of PD. However, due to its non-continuous, pulsatile delivery of L-DOPA to the brain, long-term L-DOPA administration causes deleterious side effects, including L-DOPA-induced dyskinesia (LID) among other motor complications, in the majority of patients. To achieve sustained symptomatic relief without severe L-DOPA-associated motor complications, including dyskinesia, we propose that systemic delivery of genetically engineered, chromosome- integrated, and regulatable L-DOPA-producing probiotic bacteria will avoid fluctuations in plasma L-DOPA levels and provide a more consistent delivery of L-DOPA to the brain where it can be converted to a continuous supply of dopamine in the nigrostriatal pathway. Thus, we aim to systematically evaluate the treatment feasibility and efficacy of this novel microbiome-based platform for the continuous delivery of L-DOPA in relieving motor symptoms without inducing severe dyskinesia. The scientific premise of the work is supported by key preliminary data demonstrating that: 1) the genetically reengineered, chromosome-integrated, and regulatable L-DOPA-producing E. coli Nissle 1917 probiotic strain (EcNrhaL-DOPA) efficiently produce L-DOPA both in vitro and in vivo than the older plasmid-based system, and 2) oral administration of EcNL-DOPA readily colonizes the mouse gut, achieves a steady-state plasma L-DOPA level that corresponds to the clinically effective plasma level in PD patients, and increases L-DOPA and dopamine levels in the brain. To further expand our novel preliminary results, we will pursue the following specific aims: R61 phase (i) determine the dose-response effect of orally administered EcNrhaL-DOPA on gut colonization as well as its pharmacokinetic and adaptation profiles in both C57BL and MitoPark mice; R33 phase (ii) determine the therapeutic efficacy of EcNrhaL-DOPA in the MitoPark and 6-OHDA-lesioned mouse models of PD, and (iii) determine whether sustained delivery of microbial L-DOPA prevents LID in two mouse models of LID. Our novel therapeutic pipeline strategy involving chronic delivery of probiotic L-DOPA is expected to transform the dopaminergic therapeutic modalities for PD.

抽象的帕金森病 (PD) 表现出复杂而漫长的病程，首先始于非运动障碍，然后缓慢进展为轻度至中度运动缺陷，最终造成严重的黑质纹状体多巴胺能导致的运动障碍和认知能力下降。神经元损伤最终表现为主要的锥体外系运动症状，如僵硬、运动迟缓、该提案解决了当前抗震救灾面临的最大挑战之一。用多巴胺前体左旋多巴替代多巴胺治疗帕金森病目前是口服片剂。左旋多巴/卡比多巴每天 3-4 次给药仍然是最有效且耐受性良好的治疗方法，显着改善PD早期患者的运动症状和生活质量。由于 L-DOPA 向大脑非连续、脉冲式输送，长期服用 L-DOPA 会导致有害的副作用，包括左旋多巴引起的运动障碍（LID）以及其他运动并发症大多数患者获得持续的症状缓解，而没有严重的左旋多巴相关运动。并发症，包括运动障碍，我们建议系统性地传递基因工程、染色体集成且可调节的 L-DOPA 产生益生菌将避免血浆 L-DOPA 水平的波动向大脑提供更一致的左旋多巴，并在大脑中将其转化为持续供应因此，我们的目的是系统地评估治疗的可行性。这种基于微生物组的新型平台用于持续输送左旋多巴以缓解运动症状而不引起严重的运动障碍这项工作的科学前提得到了关键的支持。初步数据表明：1）基因重组、染色体整合、可调控产左旋多巴的大肠杆菌 Nissle 1917 在体外和体外均能有效地产生益生菌菌株 (EcNrhaL-DOPA) 体内比旧的基于质粒的系统更有效，并且 2) 口服 EcNL-DOPA 很容易在小鼠中定殖肠道，达到稳态血浆左旋多巴水平，相当于帕金森病的临床有效血浆水平患者，并增加大脑中的左旋多巴和多巴胺水平，以进一步扩展我们的新颖初步研究。根据结果，我们将追求以下具体目标： R61阶段（i）确定口服药物的剂量反应效应施用 EcNrhaL-DOPA 对肠道定植及其药代动力学和适应特征的影响 C57BL 和 MitoPark 小鼠；R33 阶段 (ii) 确定 EcNrhaL-DOPA 在 MitoPark 和 6-OHDA 损伤的 PD 小鼠模型，以及 (iii) 确定是否持续递送微生物左旋多巴在两种 LID 小鼠模型中预防 LID 我们的新治疗策略管道涉及长期递送益生菌 L-DOPA 有望改变 PD 的多巴胺能治疗方式。