Erythropoietin Pretreatment of Transplanted Endothelial Colony-Forming Cells Enhances Recovery in a Cerebral Ischemia Model by Increasing Their Homing Ability: A SPECT/CT Study
Erythropoietin Pretreatment of Transplanted Endothelial Colony-Forming Cells Enhances Recovery in a Cerebral Ischemia Model by Increasing Their Homing Ability: A SPECT/CT Study
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DOI：
10.2967/jnumed.115.170308
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发表时间：
2016-11-01
影响因子：
9.3
通讯作者：
Guillet, Benjamin
中科院分区：
文献类型：
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作者：
Garrigue, Philippe;Hache, Guillaume;Guillet, Benjamin
内皮集落形成细胞（ECFC）是缺血性疾病细胞治疗的有希望的候选者，因为不到10%的缺血性卒中患者适合溶栓治疗。我们以前报道，促红细胞生成素引发的ECFC增加其在体外和体内血管生成特性在小鼠后肢缺血。本研究采用SPECT/CT评价促红细胞生成素预充ECFC是否能增强其在大鼠短暂性大脑中动脉闭塞（MCAO）后再灌注的缺血部位的归巢，并增强其对血脑屏障（BBB）破坏、脑细胞凋亡和脑血流量（CBF）的保护或再生作用。研究方法：大鼠经历了1小时的MCAO，然后再灌注，然后在MCAO后1天接受静脉注射PBS（对照，n = 10）、PBS致敏的ECFC（ECFCPBS，n = 13）或促红细胞生成素致敏的ECFC（ECFCEPO，n = 10）。通过SPECT/CT评价ECFC归巢和对BBB破坏、脑细胞凋亡和CBF的影响，直至MCAO后14天。结果表示为每个半球中大脑中动脉血管化区域中同侧与对侧活动比率的中位数+/-四分位距。在第14天进行神经元存活和星形胶质细胞增殖的组织学评价。结果：促红细胞生成素预处理增加了ECFC向缺血半球的归巢（ECFCPBS，111.0% +/- 16.0%; ECFCEPO，146.5% +/- 13.3%）。BBB破坏显著减少（对照组，387% +/- 153%; ECFCPBS，151% +/- 46% [P < 0.05]; ECFCEPO，112% +/- 9% [P < 0.001]），并与ECFC归巢呈负相关（Pearson r =-0.6930，P = 0.0002）。脑细胞凋亡显著减少（对照组，161% ± 10%; ECFCpss，141% ± 9% [P < 0.05]; ECFCEPO，118% ± 5% [P < 0.001]），并与ECFC归巢呈负相关（r =-0.7251，P < 0.0001）。ECFC组CBF显著恢复，促红细胞生成素组CBF几乎完全恢复（对照组，72% +/- 2%; ECFCPBS，90% +/- 4% [P < 0.01]; ECFCEPO，99% +/- 4% [P < 0.001]），并与ECFC归巢呈正相关（r = 0.7348，P < 0.0001）。针对ECFC上的CD 146受体的免疫阻断突出了其在促红细胞生成素引发的ECFC归巢中的显著作用（ECFCEPO，147% +/-14%，n = 4;具有针对CD 146的抗体的ECFCEPO，101% +/-12%，n = 4 [P < 0.05]）。结论：在细胞移植前用促红细胞生成素引发是一种有效的策略，以放大ECFC的迁移和植入能力及其对BBB破坏、细胞凋亡和CBF的有益影响。
Endothelial colony-forming cells (ECFCs) are promising candidates for cell therapy of ischemic diseases, as less than 10% of patients with an ischemic stroke are eligible for thrombolysis. We previously reported that erythropoietin priming of ECFCs increased their in vitro and in vivo angiogenic properties in mice with hindlimb ischemia. The present study used SPECT/CT to evaluate whether priming of ECFCs with erythropoietin could enhance their homing to the ischemic site after transient middle cerebral artery occlusion (MCAO) followed by reperfusion in rats and potentiate their protective or regenerative effect on blood-brain barrier (BBB) disruption, cerebral apoptosis, and cerebral blood flow (CBF). Methods: Rats underwent a 1-h MCAO followed by reperfusion and then 1 d after MCAO received an intravenous injection of either PBS (control, n = 10), PBS-primed ECFCs (ECFCPBS, n = 13), or erythropoietin-primed ECFCs (ECFCEPO, n = 10). ECFC homing and the effect on BBB disruption, cerebral apoptosis, and CBF were evaluated by SPECT/CT up to 14 d after MCAO. The results were expressed as median +/- interquartile range for ipsilateral-to-contralateral ratio of the activity in middle cerebral artery-vascularized territories in each hemisphere. Histologic evaluation of neuronal survival and astrocytic proliferation was performed on day 14. Results: Erythropoietin priming increased homing of ECFCs to the ischemic hemisphere (ECFCPBS, 111.0% +/- 16.0%; ECFCEPO, 146.5% +/- 13.3%). BBB disruption was significantly reduced (control, 387% +/- 153%; ECFCPBS, 151% +/- 46% [P < 0.05]; ECFCEPO, 112% +/- 9% [P < 0.001]) and correlated negatively with ECFC homing (Pearson r = -0.6930, P = 0.0002). Cerebral apoptosis was significantly reduced (control, 161% +/- 10%; ECFCpss, 141% +/- 9% [P < 0.05]; ECFCEPO,118% +/- 5% [P < 0.001]) and correlated negatively with ECFC homing (r = -0.7251, P < 0.0001). CBF was significantly restored with ECFCs and almost totally so with erythropoietin priming (control, 72% +/- 2%; ECFCPBS, 90% +/- 4% [P < 0.01]; ECFCEPO, 99% +/- 4% [P < 0.001]) and correlated positively with ECFC homing (r = 0.7348, P < 0.0001). Immunoblocking against the CD146 receptor on ECFCs highlighted its notable role in ECFC homing with erythropoietin priming (ECFCEPO, 147% +/- 14%, n = 4; ECFCEPO with antibody against CD146, 101% +/- 12%, n = 4 [P < 0.05]). Conclusion: Priming with erythropoietin before cell transplantation is an efficient strategy to amplify the migratory and engraftment capacities of ECFCs and their beneficial impact on BBB disruption, apoptosis, and CBF.