Therapeutic angiogenesis by transplantation of bone marrow stem cells

日本多中心研究(TACT-1研究组)在103例肢体缺血患者中建立了自体骨髓单个核细胞移植治疗性血管生成(BM-MNC)。安全性和有效性通过两年的随访研究(22例患者)得到证实。CD34~+细胞表达血管生成素-1，而CD34~+细胞主要表达其受体。在植入BM-MNC的肢体中，踝臂压力指数从基线的0.57增加到第4周的0.66和第24周的0.64(P&lt；0.0001)。84例患者中有70例肢体缺血后静息疼痛消退，疼痛受限的跑步机行走时间显著改善(从基线的1.4min降至第4周的3.6min和第24周的4.2min，P&lt；0.0001)。73条肢体中54条肢体缺血性溃疡或坏疽愈合，保肢成功。免疫组织化学分析显示新生毛细血管形成显着增加，Ki-67阳性的内皮细胞正在增殖。在Tact-1研究和猪模型临床前实验结果的基础上，我们对3例非选择性缺血冬眠心肌患者(n=3)进行了基于导管的BM-MNC植入的单一细胞治疗的临床试验。用27G针经导管将BM-MNC 0.2ml注入20个不同部位(共1×10^8个细胞)。局部灌注(SPECT、NOGA脑室造影)后，缺血区心绞痛发生率明显减少，室壁运动改善。连续1年半，24 h动态心电图均未发现实质性心律失常。因此，基于细胞的研究治疗缺血性心脏病被证明是安全和可行的。

Therapeutic angiogenesis by transplantation of autologous bone marrow mononuclear cells(BM-MNC) was established in 103 patients with ischemic limbs by multi-center study(TACT-1 Study Group) in Japan. Safety and efficacy was approved with 2-year follow-up study(22 patients). CD34^+ cells expressed bFGF>>VEGF>angiopoietin-1, while CD34^+ cells predominantly expressed their receptors. In BM-MNC-implanted limbs, ankle-brachial pressure index(ABI) was increased from 0.57 at baseline to 0.66 at week 4 and 0.64 at week 24 (P<0.0001). Rest pain in ischemic limbs was regressed in 70 of 84 patients, and their pain-limited treadmill walking time was significantly improved(from 1.4 min at baseline to 3.6 at week 4 and 4.2 at week 24,P<0.0001). Ischemic ulcers or gangrenes were healed in 54 of 73 limbs including successful limb salvage. Immunohistochemical analysis revealed a striking increase in new capillary formation with Ki-67 positive proliferating endothelial cells. On basis of the results of TACT-1 Study and pre-clinical experiments using porcine models, we performed the clinical trial of a sole cell therapy using catheter-based BM-MNC implantation in three no-option patients(n=3) with ischemic hibernating myocardium. 0.2 ml of BM-MNC was injected into 20 different sites(total 1x10^8 cells) via catheter with a 27G needle. Angina occurrence decreased dramatically and improvement in wall motion was observed in ischemic area with regional perfusion(SPECT, NOGA ventriography). There was no substantial arrhythmia on 24 h Holter recordings performed monthly for 1.5 year. Thus, cell-based investigational therapies for ischemic heart diseases were shown to be safe and feasible.