A Meta-Analysis of Stem Cell Mobilization by Granulocyte Colony-Stimulating Factor in the Treatment of Acute Myocardial Infarction

Objective This project was aimed at evaluating the safety and efficacy of granulocyte colony-stimulating factor (G-CSF) as an adjunctive therapy to the standard therapy [percutaneous coronary interventions (PCI) and conventional medication] after acute myocardial infarction (AMI). Methods A meta-analysis of randomized controlled trials (RCTs) of G-CSF as an adjunctive therapy to standard therapy versus standard therapy was performed. The endpoints were defined as (1) target-vessel restenosis, (2) cumulative cardiac events (CCEs) that were a combined endpoint of all-cause deaths, reinfarction, and target-vessel revascularization, and (3) the changes in left ventricular ejection fraction (LVEF) from baseline to follow-up. Results 320 patients were involved in 6 RCTs, of whom 160 were randomized to the G-CSF group and 160 to the control group. The follow-up period was 6.17 ± 3.49 months. There was no significant difference in the risk of target-vessel restenosis (P = 0.90) or CCEs (P = 0.59) between the two groups. When a pooled analysis of the changes in LVEF was performed with fixed-model effect, a significant heterogeneity was observed (P < 0.00001). The pooled analysis was thus conducted with random-model effect and did not show a significant improvement as compared to the control group (P = 0.34). A similar result was found in the sensitivity analysis based on five placebo-controlled trials involving 270 patients (P = 0.94). Conclusions G-CSF as an adjunctive therapy to standard therapy for patients with AMI may be safe. However, there is not much supporting evidence that this treatment could further improve LVEF. Since there are relatively few RCTs that meet the inclusion criteria and are heterogeneous in design, further research is required.     

Circulating Stem Cell Collection in Lymphoma and Myeloma after Mobilization with Cyclophosphamide and Granulocyte Colony-Stimulating Factor for Autologous Transplantation

We report the results of 72 leukapheresis procedures performed for autologous peripheral blood stem cell collection in 18 patients with lymphoma and myeloma, after combined mobilization with cyclophosphamide and granulocyte colony-stimulating factor (G-CSF). The numbers of mononuclear cells (MNCs), CD34+ cells and granulocyte-macrophage colony-forming units (CFU-GM) either in the peripheral circulation (preleukapheresis sample) or in the product obtained from leukapheresis (leukapheresis sample) were evaluated. A highly superior proportion of CD34+ cells (14-fold) and CFU-GM (5-fold) resulted from the mobilization therapy. CFU-GM and CD34+ cells were highly enriched with respect to all MNCs (relative recoveries: 2.13, range 0.3–41, and 1.08, range 0.2–8.5, respectively) due to an additional mobilization effect by the leukapheresis procedure. Also, a relatively strong linear correlation between the three different parameters was found in the leukapheresis product (CD34+:CFU-GM, r = 0.81; MNCs:CD34, r = 0.69; MNCs:CFU-GM, r = 0.75; CFU-GM:CD34+, and MNCs, r = 0.85). Our data suggest that the number of MNCs and CD34+ cells obtained after combined mobilization with cyclophosphamide and G-CSF can be used as predictor of the number of granulomonocytic progenitors.     

氨氯地平对糖尿病大鼠心肌梗死后骨髓内皮祖细胞动员及血管新生的改善作用

目的：观察氨氯地平对糖尿病大鼠心肌梗死（心梗）后骨髓内皮祖细胞(EPC)动员和血管新生障碍的改善作用以及对心功能的影响,并探讨其可能的分子机制。
　　方法：180~200g SPF级雄性Sprague-Dawley大鼠60只随机分为两部分：糖尿病大鼠（n=40）给予高脂饲料饲养四周后,腹腔注射30 mg/kg链脲佐菌素;非糖尿病大鼠(n=20)为正常饮食饲养。40只糖尿病大鼠结扎冠状动脉左前降支造成急性心梗模型。术后将大鼠随机分成对照组(n=20),每日予以0.5%羧甲基纤维素钠溶剂1ml灌胃,治疗组(n=20)每日予以氨氯地平2 mg/kg灌胃,继续高脂喂养四周。流式细胞术检测术前及术后不同时间点（第1、3、5、7、14和28 d）外周血CD45-/low+/CD133+/KDR+早期EPC数量,酶联免疫吸附法检测血浆血管内皮生长因子(VEGF)水平。CD31免疫荧光染色法评估心梗周围区血管新生情况。超声心动图评估心功能。免疫印迹法测定骨髓细胞中EPC动员相关信号通路蛋白的表达。
　　结果：外周血CD45-/low+/CD133+/KDR+ EPC水平术后峰值治疗组（第7 d,112±30/106单核细胞）较对照组（第5 d,55±10/106单核细胞）升高;血浆VEGF水平在心梗后峰值治疗组[第7 d ,（5.63±1.33）ng/L]较对照组[第5 d,(3.68±0.98) ng/L]升高;骨髓细胞蛋白激酶B与内皮型一氧化氮合酶的活化水平及基质金属蛋白酶-9的表达增加;心梗周围区新生毛细血管密度治疗组大鼠较对照组显著增加,左心室射血分数及左心室短轴缩短率明显提高。上述比较差异均有统计学意义（P<0.05~0.01）。
　　结论：氨氯地平治疗改善糖尿病大鼠缺血诱导的骨髓EPC动员障碍,缺血区血管新生以及心梗后心功能。这种作用可能通过改善VEGF/内皮型一氧化氮合酶信号通路活化而介导。     

Mobilization of Mesenchymal Stem Cells by Granulocyte Colony-stimulating Factor in Rats with Acute Myocardial Infarction

PURPOSE: Intravenous delivery of mesenchymal stem cells (MSCs), a noninvasive strategy for myocardial repair after acute myocardial infarction (MI), is limited by the low percentage of MSCs migration to the heart. The purpose of this study was to test whether granulocyte colony-stimulating factor (G-CSF) would enhance the colonization of intravenously infused MSCs in damaged heart in a rat model of acute MI. METHODS: After induction of anterior MI, Sprague-Dawley rats were randomized to receive: (1) saline (n = 9); (2) MSCs (n = 15); and (3) MSCs plus G-CSF (50 mug/kg/day for 5 consecutive days, n = 13). RESULTS: Flow cytometry revealed that G-CSF slightly increased surface CXCR4 expression on MSCs in vitro. After completion of G-CSF administration, MSCs showed a significantly lower colonization in bone marrow and a trend toward higher localization in the infarcted myocardium. At 3 months, vessel density in the infarct region of heart was significantly increased in MSCs group and trended to increase in MSCs + G-CSF group. However, echocardiographic and hemodynamic parameters, including left ventricular (LV) end-diastolic diameters, ejection fraction, and +/-dP/dt (max), were not statistically different. Morphological analysis showed that infarct size and collagen content were similar in the three groups. Immunohistochemistry revealed that the combined therapy accelerated endothelial recovery of the blood vessels in the ischemic myocardium. However, myocardial regeneration resulting from MSCs differentiation was not observed. CONCLUSIONS: G-CSF enhanced the migration of systemically delivered MSCs from bone marrow to infarcted heart. However, the beneficial effect of this kind of migration is limited, as cardiac function did not improve.     

同种异体骨髓间充质干细胞移植对扩张型心肌病心功能衰竭大鼠左心室功能的影响

目的探讨同种异体大鼠骨髓间充质干细胞移植在阿霉素诱导的扩张型心肌病心功能衰竭大鼠心脏内存活、分化的情况及对左心室功能的影响。方法雌性Wistar大鼠55只,随机分成正常对照组(n=10)、模型组(n=15)、诱导前移植组(n=15)和诱导后移植组(n=15)。体外分离培养雄性大鼠骨髓间充质干细胞,传至一代后用10μmol/L5-氮胞苷诱导4周,DAPI标记后,诱导前移植组移植诱导前的骨髓间充质干细胞,诱导后移植组移植诱导后的骨髓间充质干细胞;4周后检测左心室功能及移植细胞存活、分化情况。结果异体骨髓间充质干细胞移植4周后可存活并分化成心肌细胞,表达心肌细胞特异性蛋白;与模型组比较,细胞移植组左心室功能明显改善,且两细胞移植组间差异无显著性。结论同种异体骨髓间充质干细胞移植4周后可存活并分化成心肌细胞,对扩张型心肌病心功能衰竭大鼠的左心室功能有保护作用。     

Marginal Benefit/Disadvantage of Granulocyte Colony-Stimulating Factor Therapy After Autologous Blood Stem Cell Transplantation in Children: Results of a Prospective Randomized Trial

In this prospective trial, a total of 74 children who were scheduledto undergo high-dose chemotherapy followed by autologous peripheralblood stem cell transplantation (PBSCT) were prospectively randomizedat diagnosis to evaluate the effectiveness of exogenous granulocytecolony-stimulating factor (G-CSF) treatment in accelerating hematopoietic recovery after PBSCT. The diagnosis included acute lymphoblastic leukemia (ALL) (n = 27), neuroblastoma (n = 29), andmiscellaneous solid tumors (n = 18). Eligibility criteria included(1) primary PBSCT, (2) chemotherapy-responsive disease, and (3)collected cell number >1 × 10⁵ colony-formingunit-granulocyte-macrophage (CFU-GM)/kg and >1 × 10⁶CD34⁺ cells/kg patient's body weight. After applying theabove criteria, 11 patients were excluded due to disease progressionbefore PBSCT (n = 6) or a low number of harvested cells (n = 5),leaving 63 patients for analysis; 32 patients in the treatment group(300 µg/m² of G-CSF intravenously over 1 hour from day 1 of PBSCT) and 31 in the control group without treatment. Two distinctdisease-oriented high-dose regimens without total body irradiationconsisted of the MCVAC regimen usingranimustine (MCNU, 450 mg/m²), cytosinearabinoside (16 g/m²), etoposide (1.6 g/m²),and cyclophosphamide (100 mg/kg) for patients with ALL, and theHi-MEC regimen using melphalan (180 mg/m²),etoposide (1.6 g/m²), and carboplatinum (1.6 g/m²) for those with solid tumors. Five patients (two inthe treatment group and three in the control group) were subsequentlyremoved due to protocol violations. All patients survived PBSCT. Themedian numbers of transfused mononuclear cells (MNC),CD34⁺ cells, and CFU-GM were, respectively, 4.5 (range, 1 to 19) × 10⁸/kg, 8.0 (1.1 to 25) × 10⁶/kg,and 3.7 (1.2 to 23) × 10⁵/kg in the treatment group (n= 30) and 2.9 (0.8 to 21) × 10⁸/kg, 6.3 (1.1 to 34) × 10⁶/kg, and 5.5 (1.3 to 37) × 10⁵/kg,respectively, in the control group (n = 28), with no significant difference. After PBSCT, the time to achieve an absolute neutrophil count (ANC) of >0.5 × 10⁹/L in the treatment group wasless than that in the control group (median, 11 v 12 days; thelog-rank test, P = .046), although the last day of red bloodcell (RBC) transfusion (day 11 v day 10) and the duration offebrile days (>38°C) after PBSCT (4 v 4 days) wereidentical in both groups. However, platelet recovery to >20 × 10⁹/L was significantly longer in treatment group thancontrol group (26 v 16 days; P = .009) and >50 × 10⁹/L tended to take longer in the treatment group (29 v 26 days; P = .126), with significantlymore platelet transfusion-dependent days (27 v 13 days;t-test, P = .037). When patients were divided intotwo different disease cohorts, ALL patients showed no difference inengraftment kinetics between the G-CSF treatment and control groups,while differences were seen in those with solid tumors. We concludedthat the marginal clinical benefit of 1 day earlier recovery ofgranulocytes could be offset by the delayed recovery of platelets. Werecommend that the routine application of costly G-CSF therapy inchildren undergoing PBSCT should be seriously reconsidered.     

Comparison of the Seattle Heart Failure Model and Cardiopulmonary Exercise Capacity for Prediction of Death in Patients With Chronic Ischemic Heart Failure and Intracoronary Progenitor Cell Application

Background:

Despite many therapeutic advances, the prognosis of patients with chronic heart failure (CHF) remains poor. Therefore, reliable identification of high‐risk patients with poor prognosis is of utmost importance. Cardiopulmonary exercise testing (CPET) provides important prognostic information by peak O₂ uptake (peak VO₂), maximal oxygen pulse (O₂ Pmax), O₂ uptake efficiency slope (OUES), and VE/VCO₂ slope (VE/VCO₂). A different approach for prognostic assessment is the Seattle Heart Failure Model (SHFM), which is based on clinical data and calculates the estimated annual mortality.

Hypothesis:

Comparison of the prognostic value of the Seattle Heart Failure Score and cardiopulmonary excercis testing in patients with chronic heart failure.

Methods:

One hundred fifty‐seven patients with ischemic heart failure and recent intracoronary progenitor cell application were analyzed for mortality during a follow‐up of 4 years. CPET (peak VO₂, O₂ Pmax, OUES, VE/VCO₂) was performed in all patients at baseline. The SHFM score was calculated for every patient, with data obtained at the time of CPET.

Results:

During follow‐up, 24 patients died (15.2%). Nonsurvivors had significantly worse initial CPET results and a higher SHFM score compared to survivors. Receiver operating characteristics curve analysis of sensitivity and specificity revealed the largest area under the curve value for the SHFM score, followed by VE/VCO₂ slope. Kaplan Meier analysis using cutoff points of SHFM and VE/VCO₂ slope with highest sensitivity and specificity resulted in significant discrimination of survivors and nonsurvivors. By multivariate analysis, only the SHFM score persisted as independent predictor of mortality in these patients.

Conclusions:

These data indicate superior prognostic power of the SHFM score compared to CPET in patients with chronic ischemic heart failure. The authors have no funding, financial relationships, or conflicts of interest to disclose.     

慢性心力衰竭患者外周血单个核细胞核转录因子κB/p65的表达及白细胞介素-10的影响

目的探讨慢性心力衰竭(CHF)患者外周血单个核细胞(PBMCs)核转录因子κappaB(NF-κB)/p65的表达及白细胞介素-10(IL-10)的干预作用.方法对15例心力衰竭患者和15例健康对照者分离出的外周血单个核细胞进行培养,并将心力衰竭患者和健康对照者培养的外周血单个核细胞分成3组空白对照组、脂多糖(LPS)组和白细胞介素-10组,共培养24小时,对3组培养后的外周血单个核细胞NF-κB/p65的表达进行免疫组化染色检测和半定量分析.结果心力衰竭患者外周血单个核细胞NF-κB/p65胞核染色阳性率非常明显高于健康对照者[(22±8)%vs(9±2)%,P＜0.01].脂多糖组,心力衰竭患者和健康对照者外周血单个核细胞NF-κB/p65胞核染色阳性率与空白对照组比较均极显著增加(P＜0.01);而在白细胞介素-10组,心力衰竭患者和健康对照者外周血单个核细胞NF-κB/p65胞核染色阳性率与脂多糖组比较则极显著下降(P＜0.01).结论心力衰竭患者NF-κB/p65表达增高,而白细胞介素-10可下调脂多糖刺激下NF-κB/p65表达增高.