Ex vivo pretreatment of bone marrow mononuclear cells with endothelial NO synthase enhancer AVE9488 enhances their functional activity for cell therapy
Ex vivo pretreatment of bone marrow mononuclear cells with endothelial NO synthase enhancer AVE9488 enhances their functional activity for cell therapy
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DOI：
10.1073/pnas.0604144103
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发表时间：
2006-09-26
影响因子：
11.1
通讯作者：
Dimmeler, Stefanie
中科院分区：
文献类型：
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作者：
Sasaki, Ken-ichiro;Heeschen, Christopher;Dimmeler, Stefanie
缺血性心肌病患者的骨髓单个核细胞（BMC）在体内表现出较低的新生血管能力。NO在新生血管形成中起重要作用，并且NO生物利用度通常在患有血管炎的患者中降低。我们研究了用新型内皮型一氧化氮合酶（eNOS）转录增强剂AVE 9488（AVE）预处理是否可以恢复受损的血管新生能力。体外预处理AVE患者的BMC显著增加eNOS mRNA表达2.1倍（P < 0.05），ESR评估的eNOS活性增加> 3倍（P < 0.05）。eNOS表达的增加与体外迁移能力的增强（P < 0.01）和体内缺血后肢模型中灌注的BMC的新生血管能力的改善（P < 0.001）相关。灌注AVE预处理的BMC后，缺血肢体灌注改善，游泳时间增加（P < 0.05）。用eNOS抑制剂N-G-硝基-L-精氨酸甲酯进行离体预处理，可消除AVE处理的BMC对肢体灌注的增强作用。一致的是，AVE没有表现出对来自eNOS缺陷型小鼠的BMC的迁移能力受损的影响，记录了NO的具体参与。来自ENO患者的BMC的新血管形成能力降低可能会限制其在细胞治疗研究中的治疗潜力。在这里，我们表明，药理学增强eNOS表达与AVE至少部分逆转受损的功能活性的BMC从MDS患者，突出的关键作用NO祖细胞功能。
Bone marrow mononuclear cells (BMC)from patients with ischemic cardiomyopathy (ICMP) show a reduced neovascularization capacity in vivo. NO plays an important role in neovascularization, and NO bioavailability is typically reduced in patients with ICMP. We investigated whether the impaired neovascularization capacity of ICMP patient-derived progenitor cells can be restored by pretreatment with the novel endothelial NO synthase (eNOS) transcription enhancer AVE9488 (AVE). Ex vivo pretreatment of BMC from patients with ICMP with AVE significantly increased eNOS mRNA expression by 2.1-fold (P < 0.05) and eNOS activity as assessed by ESR by > 3-fold (P < 0.05). The increased eNOS expression was associated with an enhanced migratory capacity in vitro (P < 0.01) and improved neovascularization capacity of the infused BMC in an ischemic hind limb model in vivo (P < 0.001). The improvement in ischemic limb perfusion after infusion of AVE-pretreated BMC resulted in an increase in swimming time (P < 0.05). The enhancement of limb perfusion by AVE-treated BMC was abrogated by ex vivo pretreatment with the eNOS inhibitor N-G-nitro-L-arginine methyl ester. Consistently, AVE showed no effect on the impaired migratory capacity of BMC derived from eNOS-deficient mice, documenting the specific involvement of NO. The reduced neovascularization capacity of BMC from patients with ICMP may limit their therapeutic potential in cell therapy studies. Here, we show that pharmacological enhancement of eNOS expression with AVE at least partially reverses the impaired functional activity of BMC from ICMP patients, highlighting the critical role of NO for progenitor cell function.