Angiogenic Cell Therapy by Bone Marrow-Derived Monocyte-lineage Stem Cells

骨髓来源的单核细胞谱系干细胞的血管生成细胞疗法

• 批准号: 15390254
• 负责人: MATSUBARA Hiroaki
• 金额: $ 9.6万
• 依托单位: Kyoto Prefectural University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390254/
• 关键词: endothelium; angioplasty; endothelial progenitor cell; bone marrow; reendothelialization; angiogenesis; 血管再生; 骨髄移植; 単球

Peripheral blood(PB)-derived CD14^+ monocytes were shown to trans-differentiate into endothelial cell(EC)-lineage cells and contribute to neovascularization. We investigated whether bone marrow(BM)- or PB-derived CD34^-/CD14^+ cells are involved in reendothelialization after carotid balloon injury. Human BM-derived CD34^-/CD14^+ monocyte-lineage cells (BM-MLCs), but not PB-derived CD34^-CD14^+ monocytes, expressed EC-specific markers (Tie2, CD31, VE-cadherin, Endoglin) to an extent identical to mature EC. When BM-MLCs were cultured with VEGF, hematopoietic markers were drastically decreased and new EC-specific markers (Flk and CD34) were induced. BM-MLCs were intra-arterially transplanted into balloon-injured arteries of athymic nude rats. Only when BM-MLCs were activated by MCP-1 in vivo or in vitro, they adhered onto injured endothelium, differentiated into EC-like cells by losing hematopoietic markers and inhibited neointimal hyperplasia. Such MCP-dependent adhesion was not observed in PB-derived CD34^-/CD14^+ monocytes. Regenerated endothelium exhibited cobble-stone appearance, blocked extravasation of dye and induced NO-dependent vasorelaxation. Basal adhesive activities on HUVEC under laminar flow and beta-1-integrin expression (basal and active forms) were significantly increased in BM-MLCs compared with PB-derived monocytes. MCP-1 markedly enhanced adhesive activity of BM-MLCs (2.8 fold) on HUVEC by activating beta-1-integrin conformation. Thus, BM-MLCs can function as EC progenitors and acquire the ability to adhere on injured endothelium in a MCP-1-dependent manner, leading to reendothelialization followed by inhibition of intimal hyperplasia. This will open a novel window to MCP-1-mediated biological actions as well as vascular regeneration strategies using BM cell therapy.

外周血（PB）来源的CD 14 ^+单核细胞被证明转分化为内皮细胞（EC）谱系细胞，并有助于新生血管形成。我们研究了颈动脉球囊损伤后骨髓（BM）或外周血来源的CD 34 ^-/CD 14 ^+细胞是否参与了再内皮化。人骨髓来源的CD 34 ^-/CD 14 ^+单核细胞系细胞（BM-MLCs）表达的EC特异性标志物（Tie 2、CD 31、VE-钙粘蛋白、内皮糖蛋白）与成熟EC的表达程度相同，而人外周血来源的CD 34 ^-CD 14 ^+单核细胞则不表达。当BM-MLCs与VEGF一起培养时，造血标志物急剧减少，并诱导新的EC特异性标志物（Flk和CD 34）。将BM-MLCs动脉内移植到无胸腺裸大鼠球囊损伤的动脉中。BM-MLCs只有在体内或体外被MCP-1激活后才能粘附于损伤的内皮细胞上，通过失去造血标志物分化为EC样细胞，抑制新生内膜增生。在PB来源的CD 34 ^-/CD 14 ^+单核细胞中未观察到这种MCP依赖性粘附。再生的内皮呈现鹅卵石样外观，阻断染料外渗并诱导NO依赖性血管舒张。与PB来源的单核细胞相比，BM-MLCs在层流和β-1-整合素表达（基础和活性形式）下对HUVEC的基础粘附活性显著增加。MCP-1通过激活β-1-整合素构象，显著增强BM-MLCs对HUVEC的粘附活性（2.8倍）。因此，BM-MLC可以作为EC祖细胞发挥功能，并以MCP-1依赖性方式获得粘附在受损内皮上的能力，导致再内皮化，随后抑制内膜增生。这将为MCP-1介导的生物学作用以及使用BM细胞治疗的血管再生策略打开新的窗口。

项目成果

Angiogenesis by Implantation of Peripheral Blood Mononuclear Cells and Platelets into Ischemic Limbs.

通过将外周血单核细胞和血小板植入缺血肢体进行血管生成。

• 发表时间: 2002
• 期刊: Circulation 106
• 作者: Iba O;Matsubara H;Nozawa Y;PhD;Soichiro Fujiyama S;Amano K;Mori Y;Kojima H;Iwasaka T.
• 通讯作者: Iwasaka T.

Mechanism for IL-1β-mediated neovascularization unmasked by IL-1β knock-out mice, Journal of Molecular and Cellular Cardiology

IL-1β 敲除小鼠揭示 IL-1β 介导的新生血管形成机制，《分子与细胞心脏病学杂志》

• 发表时间: 2004
• 期刊: J Mol Cell Cardiol 36(4)
• 作者: Amano K;Matsubara H他6名
• 通讯作者: Matsubara H他6名

Carbon dioxide-rich water bathing enhances collateral blood flow in ischemic hindlimb via mobilization of endothelial progenitor cells and activation of NO-cGMP system

• DOI: 10.1161/01.cir.0000159329.40098.66
• 发表时间: 2005-03-29
• 期刊: CIRCULATION
• 影响因子: 37.8
• 作者: Irie, H;Tatsumi, T;Matsubara, H
• 通讯作者: Matsubara, H

Therapeutic angiogenesis for patients with limb ischemia by autologous transplantation of bone marrow cells : a pilot study and a randomised controlled trial

通过自体骨髓细胞移植治疗肢体缺血患者的血管生成：一项初步研究和一项随机对照试验

• 发表时间: 2002
• 期刊: Lancet 360
• 作者: Tateishi-Yuyama E;Matsubara H*;Murohara T;Ikeda U;Shintani S;Masaki H;Kishimoto;Y;Yoshimoto K;Akashi H;Shimada K;Iwasaka T;Imaizumi T.
• 通讯作者: Imaizumi T.

Autologous bone-marrow mononuclear cell implantation improves endothelium-dependent vasodilation in patients with limb ischemia

• DOI: 10.1161/01.cir.0000121427.53291.78
• 发表时间: 2004-03-16
• 期刊: CIRCULATION
• 影响因子: 37.8
• 作者: Higashi, Y;Kimura, M;Yoshizumi, M
• 通讯作者: Yoshizumi, M