Acute promyelocytic leukemia derived extracellular vesicles conserve PML-RARα transcript from storage-inflicted degradation: a stable diagnosis tool in APL patients

Ann Hematol. 2021 Sep;100(9):2241-2252. doi: 10.1007/s00277-021-04579-9. Epub 2021 Jul 8.

Abstract

The early death, which is more common in acute promyelocytic leukemia (APL) patients rather than other types of acute myelocytic leukemia (AML) highlights the importance of appropriate diagnostic method for early detection of this disease. The low sensitivity of the conventional methods, low tumor burden in some patients, and the need for bone marrow sampling are some of the diagnostic challenges on the way of proper detection of APL. Given these, we aimed to compare the efficacy of extracellular vesicles (EVs), as a diagnostic tool, with the existing methods. RT-PCR, qPCR, and flow cytometry were applied on EVs and their corresponding associated cellular component collected from 18 APL new cases, 23 patients with minimal residual disease (MRD), and NB4 cell line. RT-PCR results were positive in both cellular and vesicular components of all new cases, NB4 cells, and EVs in contrary to MRD cases. Normalized copy numbers (NCN) of PML-RARα were 5100 and 3950 for cell and EVs, respectively (p < 0.05). There was a significant difference in the NCN of PML-RARα between cells and EVs in BM samples. Investigating the effect of storage at room temperature revealed that PML-RARα level was retained near to the baseline level in EVs, but there was a significant reduction in its copy number in the cellular component during 7 days. Taken together, given to the acceptable stability, EVs could be introduced as a non-invasive liquid biopsy that alongside existing methods could remarkably change the paradigm of APL diagnostic approaches.

Keywords: Acute promyelocytic leukemia; Extracellular vesicle; Liquid biopsy; Normal copy number.

MeSH terms

  • Adult
  • Aged
  • Cell Line, Tumor
  • Extracellular Vesicles / genetics*
  • Female
  • Gene Dosage
  • Humans
  • Leukemia, Promyelocytic, Acute / diagnosis*
  • Leukemia, Promyelocytic, Acute / genetics
  • Male
  • Middle Aged
  • Neoplasm, Residual / diagnosis
  • Neoplasm, Residual / genetics
  • Oncogene Proteins, Fusion / genetics*
  • Promyelocytic Leukemia Protein / genetics*
  • Retinoic Acid Receptor alpha / genetics*

Substances

  • Oncogene Proteins, Fusion
  • Promyelocytic Leukemia Protein
  • RARA protein, human
  • Retinoic Acid Receptor alpha
  • promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein
  • PML protein, human