Preclinical Hemophilia Gene Therapy Study with Non-human Primates

非人类灵长类动物的临床前血友病基因治疗研究

• 批准号: 20591155
• 负责人: MIMURO Jun
• 金额: $ 2.91万
• 依托单位: Jichi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20591155/
• 关键词: 血友病; 遺伝子治療; 細胞移植; 第VIII因子; 第IX因子; 第IX

The Hemophilia gene therapy study was carried out according to the research plan. We have completed the FIX gene expression study with adeno-associated virus (AAV) vectors in mice for hemophilia B gene therapy and the hemophilia B gene therapy preclinical study is currently conducted with macaques. The AAV8 vector carrying the macaque FIX T262A was effective to express macaque FIX T262A to the therapeutic level in anti-AAV8 antibody-negative macaques whereas the vector was unable to express macaque FIX T262A to the therapeutic level in anti-AAV8 antibody-positive macaques. The novel vector-injection method for evading the inhibitory effect of anti-AAV8 antibody was developed and was effective to express the transgene-derived macaque FIX T262A in anti-AAV8 antibody-positive macaques. Human FVIII could be expressed with the AAV8 vector to the therapeutic level in FVIII deficient mice. A novel hemophilia A model animal (cloned hemophilia A pig) was generated by nuclear transfer cloning. The efficient and site specific integration of FVIII gene into the chromosome 19 was achieved with the AAV2 Rep gene. New monoclonal antibodies raised against B domain-deleted human FVIII were developed and used to quantify B domain deleted human FVIII specifically.

根据研究计划开展血友病基因治疗研究。我们已经用腺相关病毒（AAV）载体在小鼠中完成了用于血友病B基因治疗的FIX基因表达研究，目前正在猕猴中进行血友病B基因治疗的临床前研究。携带FIX T262A的AAV8载体在抗AAV8抗体阴性的猕猴体内能有效表达FIX T262A至治疗水平，而在抗AAV8抗体阳性的猕猴体内不能表达FIX T262A至治疗水平。建立了逃避抗aav8抗体抑制作用的新型载体注射方法，可在抗aav8抗体阳性的猕猴体内有效表达转基因猕猴FIX T262A。AAV8载体可在FVIII缺陷小鼠体内表达人FVIII至治疗水平。采用核转移克隆技术制备了一种新型血友病A模型动物（克隆血友病A猪）。利用AAV2 Rep基因实现了FVIII基因在19号染色体上的高效位点特异性整合。制备了新的针对B结构域缺失的人FVIII的单克隆抗体，并用于特异性定量B结构域缺失的人FVIII。

项目成果

異常第VIII因子遺伝子正常化による血友病A遺伝子治療の試み

通过使异常因子 VIII 基因正常化来尝试血友病 A 基因治疗

• 发表时间: 2008
• 作者: Mizukami;H.;柏倉裕志
• 通讯作者: 柏倉裕志

Impacto of acute cellular rejection on coagulation and f ibrinolysis biomarkers within the immediate post-operative reriod in pediatric liver transplantation.

小儿肝移植术后即刻急性细胞排斥反应对凝血和纤溶生物标志物的影响。

• 发表时间: 2010
• 期刊: Pediatr Transplant 14(3)
• 作者: Mimuro J.;Mizuta K.;Kawano Y.;Hishikawa S.;Hamano A.;Kashiwakura Y.;Ishiwata A.;Ohmori T.;Madoiwa S.;Kawarasaki H.;Sakata Y.
• 通讯作者: Sakata Y.

Vinculin activates inside-out signaling of integrin alphaIIbβ3 in Chinese hamster ovary cells.

Vinculin 激活中国仓鼠卵巢细胞中整合素 αIIbβ3 的由内而外信号传导。

• 发表时间: 2010
• 期刊: Biochem Biophys Res Commun 400(3)
• 作者: Ohmori T.;Kashiwakura Y.;Ishiwata A,. Madoiwa S.;Mimuro J.;Honda S.;Miyata T.;Sakata Y.
• 通讯作者: Sakata Y.

The chicken hypersensitive site-4 chromatin insulator sequence protects clonal domina of hematopoietic stem cells transduced with a self-inactivating SIV vector in platelet-directed gene therapy.

在血小板定向基因治疗中，鸡过敏位点 4 染色质绝缘体序列可保护用自失活 SIV 载体转导的造血干细胞的克隆域。

• 发表时间: 2010
• 作者: Tsukasa Ohmori;Yuji Kashiwakura;Akira Ishiwata;Seiji Madoiwa;Eiji Akiba;Mamoru Hasegawa;Jun Mimuro;Keiya Ozawa;Yoichi Sakata
• 通讯作者: Yoichi Sakata

非ヒト霊長類を用いた血友病A伝子治療研究に向けたヒトBDDFVIII特異的検出法の確立

建立人类BDDFVIII特异性检测方法，用于非人灵长类血友病A基因治疗研究

• 发表时间: 2010
• 作者: 柏倉裕志;三室淳;石渡彰;安本篤史;坂田飛鳥;大森司;窓岩清治;水上浩明;小野文子;小澤敬也;坂田洋一
• 通讯作者: 坂田洋一