Identification and isolation of homogenous AML derived extracellular vesicles for MRD detection in pediatric AML

AML specific mutations were previously detected by our group in EV-DNA employing ultracentrifugation (UC) based EV isolation either at diagnosis or during therapy. However, it is challenging to detect minimal residual disease (MRD) status in these samples because the UC method provides heterogeneous EVs containing DNA from leukemic and non-leukemic immune and stromal cells, leading to a massive decline in mutational information after remission during MRD stages. Therefore, there is an urgent need for novel strategies to be developed to allow homogenous EV preparation with high purity.

To obtain homogenous EVs with biomarker potential, we have established a single EV identification and capturing platform using antigen panels specific for immunocapturing AML-cell-derived EVs. Using the cohort of patient samples at diagnosis (n=30) and three different time points of therapy (n=120), we aimed to perform EV isolation using AML-EVs surface antibody bead capturing approach. Next, after the isolation of EV-DNA, MRD detection will be performed utilizing pediatric AML NGS panel to establish EV biomarker potential in leukemia diagnostics.

Publication History

Article published online:
17 May 2022