Basic and clinical research for the application of "Ex vivo expanded erythroblast transplantation"

“体外扩大成红细胞移植”应用的基础与临床研究

• 批准号: 17590714
• 负责人: TOBA Ken
• 金额: $ 2.3万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590714/
• 关键词: Angiogenesis; Erythroblasts; Erythropoietin; Ex vivo expansion; Cell Therapy

Autologous bone marrow cell implantation (BMI) has been utilized for the treatment of limb ischemia, however the mechanism of angiogenesis by the therapy is not well known. As large as 500 to 1,000 mL of bone marrow collection is usually required for the treatment. The aim of this study was to develop a sophisticated cell therapy by using ex vivo cell culture technique.In the beginning, we studied the mechanism of angiogenesis by BMI. We expected that the angiogenesis by the therapy mimicked the constitutional angiogenesis in the bone marrow tissue. Several angiogenic factors were strongly produced by erythroblasts. In vitro angiogenesis of HUVEC was conducted by erythroid colonies but not by myeloid colonies prepared from human blood stem cells. Erythroid cells in bone marrow played essential roles for angiogenesis in mouse limb ischemia model. In the in vivo model, simultaneous administration of erythropoietin with BMI enhanced angiogenesis presumably through surviving effect on erythroid cells extramedullary.Next, we developed an ex vivo expansion system of erythroid cells from bone marrow stem cells. As the results, only 1 mL of bone marrow was enough to produce angiogenic factor producing cells that were fully compatible with 500 to 1,000 mL of freshly harvested bone marrow. The ex vivo expanded erythroblasts also expressed strong angiogenic effect in the mouse limb ischemic model.A phase I/II trial of "Ex vivo expanded autologous erythroblast transplantation for the treatment of the patients with limb ischemia (EVEETA)" has been approved by the ethical committee and the IRB in the hospital. Cell culture for the clinical use will be enforced in the GMP-grade Cell Processing Center (CPC) in the hospital.

自体骨髓细胞移植（BMI）已被用于治疗肢体缺血，但其血管生成机制尚不清楚。治疗通常需要采集多达500至1，000 mL的骨髓。本研究的目的是利用体外细胞培养技术开发一种成熟的细胞治疗方法。我们预期通过治疗的血管生成模拟骨髓组织中的组成性血管生成。成红细胞强烈产生几种血管生成因子。HUVEC的体外血管生成由红系集落进行，而不是由人造血干细胞制备的髓系集落进行。骨髓红系细胞在小鼠肢体缺血模型血管生成中起重要作用。在体内模型中，促红细胞生成素与BMI同时给药可能通过对髓外红系细胞的存活作用增强血管生成。接下来，我们开发了一种从骨髓干细胞中扩增红系细胞的体外系统。结果，仅1 mL骨髓就足以产生与500至1，000 mL新鲜采集的骨髓完全相容的血管生成因子产生细胞。体外扩增的成红细胞在小鼠肢体缺血模型中也表现出较强的血管生成作用。“体外扩增的自体成红细胞移植治疗肢体缺血患者（EVEETA）”的I/II期临床试验已获得该院伦理委员会和IRB的批准。临床使用的细胞培养将在医院的GMP级细胞处理中心（CPC）进行。

项目成果

Erythroid cells play essential roles in angiogenesis by bone marrow cell implantation

红细胞在骨髓细胞植入的血管生成中发挥重要作用

• 发表时间: 2006
• 期刊: J Mol Cell Cardiol 40
• 作者: Ozawa T;Toba K;Kato K;et. al.
• 通讯作者: et. al.

骨髄単核細胞移植を用いた血管新生治療の作用機所とエリスロポエチンの臨床応用

骨髓单个核细胞移植血管生成治疗的作用机制及促红细胞生成素的临床应用

• 发表时间: 2006
• 期刊: 脈管学 46・5
• 作者: 加藤公則;鳥羽健;小田雅人;他
• 通讯作者: 他

Marked decrease of plasma VEGF after implantation of autologous bone marrow mononuclear cells in a patient with critical limb ischemia-a case report

肢体严重缺血患者自体骨髓单个核细胞植入后血浆VEGF明显下降一例报告

• 发表时间: 2006
• 期刊: Angiology 57(2)
• 作者: Ozawa;T;Kato;K;Sanada;H;Makiyama;Y;Saigawa;T;Souda;S;Hashimoto;S;Furukawa;T;Toba;K;Kodama;M;Fujiwara;H;Namura;O;Hayashi;J;Yoshimura;N;Aizawa;Y
• 通讯作者: Y

骨髄単核細胞移植を用いた血管新生療法の作用機序とエリスロポエチンの臨床応用

骨髓单个核细胞移植血管生成治疗的作用机制及促红细胞生成素的临床应用

• 发表时间: 2006
• 期刊: 脈管学 46・5
• 作者: 加藤公則;鳥羽健;小田雅人
• 通讯作者: 小田雅人

In vitro effect of cyclosporin A, mitomycin C and prednisolone on cell kinetics in cultured human umbilical vein endothelial cells

• DOI: 10.1016/j.thromres.2004.09.001
• 发表时间: 2005-01-01
• 期刊: THROMBOSIS RESEARCH
• 影响因子: 7.5
• 作者: Seki, Y;Toba, K;Aizawa, Y
• 通讯作者: Aizawa, Y