Retrovirus-mediated gene transfer as an approach to analyze neuroblastoma relapse after autologous bone marrow transplantation.
Retrovirus-mediated gene transfer as an approach to analyze neuroblastoma relapse after autologous bone marrow transplantation.
复制标题
逆转录病毒介导的基因转移作为分析自体骨髓移植后神经母细胞瘤复发的方法。
DOI：
10.1089/hum.1992.3.2-129
复制
发表时间：
1992
影响因子：
4.2
通讯作者：
Brenner,MK
中科院分区：
文献类型：
--
作者：
Rill,DR;Buschle,M;Foreman,NK;Bartholomew,C;Moen,RC;Santana,VM;Ihle,JN;Brenner,MK
播散性神经母细胞瘤是一种儿童恶性肿瘤，通常采用强化化疗/放疗后自体骨髓移植（ABMT）治疗。治疗后复发的比例很高。目前尚不清楚复发是由于患者体内残留的疾病还是由于注入骨髓中残留的恶性细胞受到污染的结果，骨髓在消融化疗/放疗之前必须收集和储存。假设注入的骨髓中残留的细胞会导致复发，因此在再注入之前采用骨髓清除。但是，既没有确定这一程序的必要性，也没有确定其效力。我们现在展示了使用LNL6载体的逆转录病毒介导的基因转移如何解决这个问题。通过观察G418中单个神经母细胞瘤细胞集落的生长情况，以及单个集落的聚合酶链反应（PCR），可以对患者骨髓中的克隆源性神经母细胞瘤细胞进行转导和NEORgene检测。转导效率在0 ~ 13.5%之间。如果骨髓在输注前暴露于LNL6，并且在复发时检测到标记细胞，这将证明输注的骨髓有助于疾病复发。这项技术可以用来分析骨髓净化技术的功效。由于来自这些患者的正常祖细胞也被标记，该技术可用于研究改变注入骨髓的重构和可转导性的因素。使用这种方法的临床研究现已开始。
Disseminated neuroblastoma is a malignancy of children often treated by intensive chemotherapy/radiotherapy followed by autologous bone marrow transplantation (ABMT). A high proportion of those treated subsequently relapse. It is unknown if relapse is a consequence of residual disease in the patient or of contaminating malignant cells remaining in the infused marrow, which, of necessity, is harvested and stored prior to ablative chemotherapy/radiotherapy. The assumption that residual cells in the infused marrow contribute to relapse has lead to the adoption of marrow purging prior to reinfusion. However, neither the necessity nor the efficacy of the procedure have been established. We now show how retroviral-mediated gene transfer using the LNL6 vector may resolve this issue. Clonogenic neuroblastoma cells in patient marrow can be transduced and the NEORgene detected by observing individual neuroblastoma cell colony growth in G418, and by polymerase chain reaction (PCR) of individual colonies. Efficiency of transduction is between 0 and 13.5%. If marrow is exposed to LNL6 prior to infusion and marked cells are detected at the time of relapse, this would demonstrate that infused marrow contributed to disease recurrence. The technique could then be used to analyze the efficacy of marrow purging techniques. Since normal progenitor cells from these patients are also marked, the technique can be used to study factors that modify reconstitution and transducibility of infused marrow. Clinical studies using this approach have now begun.