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This research team previously developed a qPCR-based system that was used to detect levels of eleven markers associated with the cancer stem cells that cause neuroblastoma reoccurrence. This method enabled them to monitor MRD in high-risk neuroblastoma patients. However, it was difficult to accurately track the changes taking place using qPCR. For the current study, the research group decided to use the more sensitive and reproducible ddPCR to evaluate MRD.
First of all, 208 bone marrow samples and 67 peripheral blood samples were taken from high- risk neuroblastoma patients, and 103 bone marrow samples and 107 peripheral blood samples were obtained from healthy individuals (control). Seven markers were selected from among the eleven markers that are highly expressed in the cancer stem cells that cause neuroblastoma regrowth/relapse. Using ddPCR, they calculated the levels of these seven markers in the bone marrow and peripheral blood samples.
The research group discovered that they could more accurately diagnose high-risk neuroblastoma patients when calculating the aggregated expression levels of all 7 markers using ddPCR, as opposed to looking at the expression level of each marker individually. The changes in the expression levels of these seven markers reflected the amount of tumors, the stage of illness (remission, stable, or progression) and the sample collection period (diagnosis, treatment, post-treatment, or relapse).
The expression levels of the 7 markers were calculated for 73 bone marrow samples obtained from post-treatment high-risk neuroblastoma patients. The expression levels of the markers in the bone marrow samples of the 17 patients who had suffered tumor relapse/regrowth was significantly higher compared to the levels in the 56 patients who hadn't. By comparing the two PCR methods using the same 73 samples, it was predicted that the expression levels of the seven markers calculated by ddPCR would be able to more accurately predict tumor relapse/regrowth than qPCR.
Based on these results, it is expected that the ddPCR monitoring method developed by this study will improve the monitoring of MRD in high-risk neuroblastoma patients, enabling the possibility of tumor relapse or regrowth to be more accurately predicted. Further developments
The results of this research suggest that it is possible to accurately monitor MRD in post-treatment high-risk neuroblastoma patients. Next, this methodology needs to be evaluated through prospective clinical studies on a larger number of patients across multiple hospitals. It is expected that this monitoring method can be used as a basis for predicting patients' prognoses and developing new treatments for high-risk neuroblastoma patients. Source:
Kobe University Journal reference:
Thwin, K.K.M., et al. (2020) Level of Seven Neuroblastoma-Associated mRNAs Detected by Droplet Digital PCR Is Associated with Tumor Relapse/Regrowth of High-Risk Neuroblastoma Patients. Journal of Molecular Diagnostics . doi.org/10.1016/j.jmoldx.2019.10.012 .
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