Primate Study for Gene Therapy of Porkinson's disease for clinic

灵长类动物波金森氏病基因治疗的临床研究

• 批准号: 14370211
• 负责人: MOCHIZUKI Hideki
• 金额: $ 8.9万
• 依托单位: Juntendo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370211/
• 关键词: Parkinson's disease; Gene therapy; AAV vector; Caspase 11; Apaf-1-DN; MPTP; アデノ随伴ウイルスベクター; α-synuclein; 猿; Apaf-1-DN; 大型動物(猿); Apaf-1; alpha-synuclein; アポトーシス

Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. In this report, we focused on the pathogenesis and treatment of PD.The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) causes a parkinsonian syndrome in humans and primates after selective uptake of its metabolite, MPP^+, into dopaminergic neurons. We sought to determine the major pathway of MPTP toxicity from among several apoptotic pathways. We already established in vivo models of the inhibition of caspase cascade using adeno-asociated virus (AAV) vectors. We showed persistent high levels of focal Apaf-1 CARD or caspase-1 C285G mutant expression were available for further in vivo studies and for possible anti-apoptotic gene therapy in patients with PD. In this report, we confirm the effect and the safe of AAV vector using the primates for the clinical trial. AAV vector, a single-strand DNA virus from the parvovirus family, has become one of the most promising vectors for the introduction of the gene of interest into neurons (Mochizuki et al.2001). With regard to safety issues, the genetically modified primate model is simple and safe since the AAV is a non-pathogenic virus. Furthermore, we have already established the procedure for preparing parkinsonian rat and mice models that overexpress mutated α-synuclein using the AAV vector system. Thus, by stereotactically injecting AAV-mutated α-synuclein in the substantia nigra of monkeys; we established a genetically modified primate model with overt hemiparkinsonism in Tukuba primate center. Further analysis will be performed. (240)

帕金森病（Parkinson's disease，PD）是仅次于阿尔茨海默病（Alzheimer's disease）的第二大神经退行性疾病。1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）是一种神经毒素，它的代谢产物MPP^+被多巴胺能神经元选择性摄取后，可引起人类和灵长类的帕金森综合征。我们试图从几种凋亡途径中确定MPTP毒性的主要途径。我们已经建立了使用腺相关病毒（AAV）载体抑制caspase级联反应的体内模型。我们发现持续高水平的局灶性Apaf-1 CARD或caspase-1 C285 G突变体表达可用于进一步的体内研究和PD患者可能的抗凋亡基因治疗。在本报告中，我们使用灵长类动物进行临床试验，以确认AAV载体的有效性和安全性。AAV载体是一种来自细小病毒科的单链DNA病毒，已成为将目的基因导入神经元的最有前途的载体之一（Mochizuki et al.2001）。关于安全性问题，由于AAV是非致病性病毒，因此遗传修饰的灵长类动物模型简单且安全。此外，我们已经建立了使用AAV载体系统制备过表达突变α-突触核蛋白的帕金森病大鼠和小鼠模型的程序。因此，通过立体定向注射AAV突变的α-突触核蛋白到猴子的黑质中，我们在Tukuba灵长类动物中心建立了具有明显偏侧帕金森症的遗传修饰灵长类动物模型。将进行进一步分析。（240）

项目成果

Furuya T, Hayakawa H, Yamada Y, Yoshimi K, Hisahara S, Miura M, Mizuno Y, Mochizuki H: "Caspase-11 mediates inflammatory dopaminergic cell death in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease."J Neurosci. 25;24(8).

Furuya T、Hayakawa H、Yamada Y、Yoshimi K、Hisahara S、Miura M、Mizuno Y、Mochizuki H：“Caspase-11 介导 1-甲基-4-苯基-1,2,3,6 中的炎症性多巴胺能细胞死亡

Suzuki A, Obi K, Urabe T, Hayakawa H, Yamada M, Kaneko S, Onodera M, Mizuno Y, Mochizuki H: "Feasibility of ex vivo gene therapy for neurological disorders using the new retroviral vector GCDNsap packaged in the vesicular stomatitis virus G protein."J Neu

Suzuki A、Obi K、Urabe T、Hayakawa H、Yamada M、Kaneko S、Onodera M、Mizuno Y、Mochizuki H：“使用包装在水泡性口炎病毒 G 中的新型逆转录病毒载体 GCDNsap 进行离体基因治疗神经系统疾病的可行性

Komine-Kobayashi M, Chou N, Mochizuki H, Nakao A, Mizuno Y, Urabe T.: "Dual role of Fcgamma receptor in transient focal cerebral ischemia in mice."Stroke. Apr. 35(4). 958-963 (2004)

Komine-Kobayashi M、Chou N、Mochizuki H、Nakao A、Mizuno Y、Urabe T.：“Fcgamma 受体在小鼠短暂性局灶性脑缺血中的双重作用。”中风。

Choi JB, Uchino H, Azuma K, Iwashita N, Tanaka Y, Mochizuki H, Migita M, Shimada T, Kawamori R, Watada H.: "Little evidence of transdifferentiation of bone marrow-derived cells into pancreatic beta cells."Diabetologia oct. 46(10). 1366-1374 (2003)

Choi JB、Uchino H、Azuma K、Iwashita N、Tanaka Y、Mochizuki H、Migita M、Shimada T、Kawamori R、Watada H.：“骨髓来源细胞转分化为胰腺 β 细胞的证据很少。”Diabetologia oct

Mochizuki H, Mizuno Y: "Gene therapy for Parkinson's disease"J Neural Transm Suppl. 65. 205-213 (2003)

Mochizuki H、Mizuno Y：“帕金森病的基因治疗”J Neural Transm Suppl。