Blood Res.  2024;59:43. 10.1007/s44313-024-00051-5.

Bone marrow mesenchymal stem cell exosomes suppress JAK/STAT signaling pathway in acute myeloid leukemia in vitro

• Jalilivand S
• Nabigol M
• Bakhtiyaridovvombaygi M
• Gharehbaghian A

• KMID: 2564323
• DOI: http://doi.org/10.1007/s44313-024-00051-5

Abstract

Introduction
Despite advances in the treatment of acute myeloid leukemia (AML), refractory forms of this malignancy and relapse remain common. Therefore, development of novel, synergistic targeted therapies are needed urgently. Recently, mesenchymal stem cells (MSCs) have been shown to be effective in treating various diseases, with most of their therapeutic outcomes attributed to their exosomes. In the current study, we investigated the effects of bone marrow mesenchymal stem cell (BM-MSC) exosomes on the expression of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling genes involved in AML pathogenesis. Material and Methods Exosomes were isolated from BM-MSCs and confirmed using transmission electron microscopy, dynamic light scattering, and flow cytometry. Subsequently, the exosome concentration was estimated using the bicinchoninic acid assay, and HL-60 cells were cocultured with 100 µg/mL of BM-MSC exosomes. Finally, the JAK2, STAT3, and STAT5 expression levels were analyzed using qRT-PCR.
Results
The exosome characterization results confirmed that most isolated nanoparticles exhibited a round morphology, expressed CD9, CD63, and CD81, which are specific protein markers for exosome identification, and ranged between 80 and 100 nm in diameter. Furthermore, qRT-PCR analysis revealed a significant downregulation of JAK2, STAT3, and STAT5 in HL-60 cells treated with 100 μg/mL of BM-MSC exosomes.
Conclusion
Since JAK/STAT signaling contributes to AML survival, our findings suggest that the downregulation of JAK/STAT genes by BM-MSC exosomes in leukemic cells may aid in designing a potent therapeutic strategy for AML treatment.

Keyword

Bone marrow mesenchymal stem cell; Exosome; Acute myeloid leukemia; JAK2; STAT3; STAT5

Figure

• 

  Fig. 1 The identification and confirmation of BM-MSCs. A In passage 2, BM-MSCs demonstrated fibroblast-like morphology under an inverted microscope. B The flow cytometry was used to check the immune phenotype of BM-MSCs. BM-MSCs lacked CD45, CD34, and CD14, but expressed CD105, CD90, and CD73

• 

  Fig. 2 The identification and confirmation of BM-MSC exosomes. A The size of the isolated exosomes was determined using the DLS technique. The exosome sizes varied from 80–100 nm. B Morphology of isolated exosomes under TEM. The BM-MSC exosomes demonstrated a spherical-shaped morphology. C The expression of the CD63, CD81, and CD9 exosome-specific surface markers in BM-MSC exosomes was examined using flow cytometry

• 

  Fig. 3 The impact of BM-MSC exosomes on the JAK/STAT gene expression of HL-60 cells. JAK2, STAT3, and STAT5 levels were notably reduced in HL-60 cells following treatment with 100 μg/mL of exosomes. The expression levels were normalized using the GAPDH housekeeping gene. Statistical significance was observed (*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001) compared to the control group (n = 3)

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