Abstract：
Every year, thousands of donors are exposed to granulocyte-colony stimulating factor (G-CSF) for stem cell mobilization in hematopoietic stem cell transplantations (HSCT). Previous studies about the genotoxicity of G-CSF were inconclusive. In this study, the genotoxic effects of G-CSF in peripheral blood stem cell (PBSC) donors were evaluated prospectively by using three different validated and reliable methods for the first time in the literature to the best of our knowledge.
Donors of PBSC transplantation (n=36), who received G-CSF were evaluated for genotoxicity by micronucleus test (MNT), nuclear division index (NDI), and comet assay (CA). Genotoxic effects are expected to cause an increase in MNT and CA values and decrease in NDI. Blood samples were collected at three timepoints (TP): before starting G-CSF (TP1), after G-CSF for five days (TP2), and one month after the last dose (TP3). Sixteen controls were included for baseline comparison of genotoxicity tests. CD34 cell counts and hemograms were also analyzed.
MNT and CA parameters; comet and tail length, tail DNA%, and tail moment, showed no change in time whereas another CA parameter, Olive`s tail moment (OTM) was increased significantly at TP3 compared to both baseline and TP2 (p=0.002 and p=0.017, respectively). Nuclear division index decreased significantly at TP2 (p < 0.001), then increased above baseline at TP3 (p=0.004). Baseline comparison with controls showed higher MN frequency in donors without statistical significance (p=0.059). Whereas, CA results were significantly higher in controls. CD34 cell count showed moderate positive correlation with white blood cell count at TP2 (Pearson R=0.495, p=0.004).
Our results showed the genotoxic effect of G-CSF in healthy donors, in two of the three tests performed, short-term effect in NDI, and long-lasting effect in OTM. So, this study provides novel information for the debate about the genotoxicity of G-CSF and supports the need for further studies with a larger sample size and longer follow-up.
Donors of PBSC transplantation (n=36), who received G-CSF were evaluated for genotoxicity by micronucleus test (MNT), nuclear division index (NDI), and comet assay (CA). Genotoxic effects are expected to cause an increase in MNT and CA values and decrease in NDI. Blood samples were collected at three timepoints (TP): before starting G-CSF (TP1), after G-CSF for five days (TP2), and one month after the last dose (TP3). Sixteen controls were included for baseline comparison of genotoxicity tests. CD34 cell counts and hemograms were also analyzed.
MNT and CA parameters; comet and tail length, tail DNA%, and tail moment, showed no change in time whereas another CA parameter, Olive`s tail moment (OTM) was increased significantly at TP3 compared to both baseline and TP2 (p=0.002 and p=0.017, respectively). Nuclear division index decreased significantly at TP2 (p < 0.001), then increased above baseline at TP3 (p=0.004). Baseline comparison with controls showed higher MN frequency in donors without statistical significance (p=0.059). Whereas, CA results were significantly higher in controls. CD34 cell count showed moderate positive correlation with white blood cell count at TP2 (Pearson R=0.495, p=0.004).
Our results showed the genotoxic effect of G-CSF in healthy donors, in two of the three tests performed, short-term effect in NDI, and long-lasting effect in OTM. So, this study provides novel information for the debate about the genotoxicity of G-CSF and supports the need for further studies with a larger sample size and longer follow-up.
摘要：
背景：每年，在造血干细胞移植(HSCT)中,成千上万的捐献者暴露于粒细胞集落刺激因子(G-CSF)以动员干细胞.先前关于G-CSF遗传毒性的研究尚无定论。在这项研究中,据我们所知,在文献中首次使用3种不同的经验证和可靠的方法前瞻性评估了G-CSF对外周血干细胞(PBSC)供者的遗传毒性作用.
方法：PBSC移植的供体（n=36），接受G-CSF的患者通过微核试验（MNT）评估遗传毒性，核分裂指数(NDI)，和彗星测定（CA）。预期遗传毒性作用会导致MNT和CA值的增加和NDI的减少。在三个时间点（TP）收集血样：开始G-CSF之前（TP1），G-CSF后五天（TP2），最后一次给药（TP3）后一个月。包括16个对照用于遗传毒性测试的基线比较。还分析了CD34细胞计数和血流量图。
结果：MNT和CA参数；彗星和尾巴长度，尾部DNA%,和尾矩，显示时间没有变化，而另一个CA参数，与基线和TP2相比，TP3时的Olive尾矩（OTM）显着增加（分别为p=0.002和p=0.017）。核分裂指数在TP2时显著下降(p<0.001)，然后在TP3增加到基线以上(p=0.004)。与对照的基线比较显示供体中更高的MN频率,无统计学显著性(p=0.059)。然而,对照组的CA结果明显较高。CD34细胞计数与TP2时的白细胞计数呈中度正相关（PearsonR=0.495，p=0.004）。
结论：我们的结果显示了G-CSF对健康供者的遗传毒性作用，在进行的三项测试中，NDI的短期效应，在OTM中的持久效果。所以,这项研究为有关G-CSF遗传毒性的争论提供了新的信息,并支持需要更大样本量和更长随访时间的进一步研究.
方法：PBSC移植的供体（n=36），接受G-CSF的患者通过微核试验（MNT）评估遗传毒性，核分裂指数(NDI)，和彗星测定（CA）。预期遗传毒性作用会导致MNT和CA值的增加和NDI的减少。在三个时间点（TP）收集血样：开始G-CSF之前（TP1），G-CSF后五天（TP2），最后一次给药（TP3）后一个月。包括16个对照用于遗传毒性测试的基线比较。还分析了CD34细胞计数和血流量图。
结果：MNT和CA参数；彗星和尾巴长度，尾部DNA%,和尾矩，显示时间没有变化，而另一个CA参数，与基线和TP2相比，TP3时的Olive尾矩（OTM）显着增加（分别为p=0.002和p=0.017）。核分裂指数在TP2时显著下降(p<0.001)，然后在TP3增加到基线以上(p=0.004)。与对照的基线比较显示供体中更高的MN频率,无统计学显著性(p=0.059)。然而,对照组的CA结果明显较高。CD34细胞计数与TP2时的白细胞计数呈中度正相关（PearsonR=0.495，p=0.004）。
结论：我们的结果显示了G-CSF对健康供者的遗传毒性作用，在进行的三项测试中，NDI的短期效应，在OTM中的持久效果。所以,这项研究为有关G-CSF遗传毒性的争论提供了新的信息,并支持需要更大样本量和更长随访时间的进一步研究.
