Novel Re-engineered L DOPA probiotic therapy for Parkinsons Disease

新型重新设计的左旋多巴益生菌疗法治疗帕金森病

• 批准号: 10453379
• 负责人: Anumantha Gounder Kanthasamy
• 金额: $ 16.23万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10453379
• 关键词: Novel; Re; engineered; DOPA; probiotic

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 帕金森氏病（PD）暴露的复杂且长时间的疾病病程首先从非 - 运动障碍，然后慢慢发展为轻度到中度电动机定义，最终导致严重 运动障碍和认知能力下降。黑质纹状体多巴胺能引起的多巴胺缺乏症 神经元损伤最终表现为刚性肌锥体室外运动症状，胸肌，肌脱节， 震颤和姿势不稳定。该提案涉及当前反抗面临的最大挑战之一 多巴胺前体L-DOPA替代多巴胺的帕金森氏症疗法。目前，口服平板电脑 L-DOPA/Carbidopa 3-4次/天的剂量仍然是最有效且耐受性良好的治疗方法 显着改善了PD早期患者的运动症状和生活质量。但是，到期 长期L-DOPA给药的L-DOPA向大脑的非连续，脉动传递 有害的副作用，包括L-DOPA诱导的运动障碍（盖）以及其他运动并发症， 大多数患者。在没有严重L-DOPA相关的电动机的情况下实现持续的症状缓解 并发症，包括运动障碍，我们提出，一般设计的，染色体的系统性输送 综合和调节的L-DOPA产生的益生菌细菌将避免在血浆L-DOPA水平上波动 并提供更一致的l-dopa向大脑的传递，可以将其转换为连续供应 在黑质途径中的多巴胺。这，我们旨在系统地评估治疗可行性 并简化这个基于微生物组的新型平台，用于持续交付L-DOPA以缓解 运动症状没有引起严重运动障碍。工作的科学前提得到了密钥的支持 初步数据表明：1）一般重新设计，染色体集成和调节 产生L-DOPA的大肠杆菌Nissle 1917益生菌菌株（ECNRHAL-DOPA）有效地产生了L-DOPA，并在体外和 体内比较旧的基于质粒的系统，而2）ECNL-DOPA的口服给予小鼠容易地分裂 肠道，达到稳态等离子体L-DOPA水平，与PD中的临床有效等离子体水平相对应 患者，并增加大脑中的L-DOPA和多巴胺水平。进一步扩大我们的小说初步 结果，我们将追求以下特定目标：R61阶段（i）确定口服的剂量反应效应 在肠道定植上管理Ecnrhal-Dopa及其药代动力学和适应谱。 C57BL和Mitopark小鼠； R33期（II）确定Mitopak中Ecnrhal-Dopa的治疗效率和 PD的6- OHDA渗透的小鼠模型，（iii）确定微生物L-DOPA是否持续递送 防止两个小鼠盖模型中的盖子。我们的新型热管道策略涉及 益生菌L-DOPA有望改变PD的多巴胺能治疗方式。