Journal
JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH
Volume 40, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s13046-020-01776-8
Keywords
Myelofibrosis; Extracellular vesicles; Inflammation; microRNAs
Categories
Funding
- SIE-Societa Italiana di Ematologia e Associazione Amici di Beat Leukemia Dr. Alessandro Cevenini ONLUS
- Associazione Italiana Ricerca sul Cancro (AIRC-Fellowship Abroad) [Rif. 20930]
- Roberto and Cornelia Pallotti Foundation [M.C3]
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In this study, it was found that the absolute numbers of circulating CD34(+) cells were significantly increased in triple-negative (TN) myelofibrosis patients. These cells displayed in vitro defective functions and were unresponsive to the inflammatory microenvironment. Furthermore, the circulating extracellular vesicles (EVs) from TN patients were found to promote the survival of TN CD34(+) cells, suggesting a distinct EV-driven signal promoting malignant hemopoiesis in TN patients. The distinct gene expression profiling analysis and microRNA signature in TN patients may provide insights for novel therapeutic approaches for this rare form of myelofibrosis.
BackgroundMyelofibrosis (MF) is a clonal disorder of hemopoietic stem/progenitor cells (HSPCs) with high prevalence in elderly patients and mutations in three driver genes (JAK2, MPL, or CALR). Around 10-15% of patients are triple-negative (TN) for the three driver mutations and display significantly worse survival. Circulating extracellular vesicles (EVs) play a role in intercellular signaling and are increased in inflammation and cancer. To identify a biomolecular signature of TN patients, we comparatively evaluated the circulating HSPCs and their functional interplay with the microenvironment focusing on EV analysis.MethodsPeripheral blood was collected from MF patients (n =29; JAK2(V617F) mutation, n =23; TN, n =6) and healthy donors (HD, n =10). Immunomagnetically isolated CD34(+) cells were characterized by gene expression profiling analysis (GEP), survival, migration, and clonogenic ability. EVs were purified from platelet-poor plasma by ultracentrifugation, quantified using the Nanosight technology and phenotypically characterized by flow cytometry together with microRNA expression. Migration and survival of CD34(+) cells from patients were also analyzed after in vitro treatments with selected inflammatory factors, i.e. (Interleukin (IL)-1 beta, Tumor Necrosis Factor (TNF)-alpha, IL6) or after co-culture with EVs from MF patients/HD.ResultsThe absolute numbers of circulating CD34(+) cells were massively increased in TN patients. We found that TN CD34(+) cells show in vitro defective functions and are unresponsive to the inflammatory microenvironment. Of note, the plasma levels of crucial inflammatory cytokines are mostly within the normal range in TN patients. Compared to JAK2(V617F)-mutated patients, the GEP of TN CD34(+) cells revealed distinct signatures in key pathways such as survival, cell adhesion, and inflammation. Importantly, we observed the presence of mitochondrial components within plasma EVs and a distinct phenotype in TN-derived EVs compared to the JAK2(V617F)-mutated MF patients and HD counterparts. Notably, TN EVs promoted the survival of TN CD34(+) cells. Along with a specific microRNA signature, the circulating EVs from TN patients are enriched with miR-361-5p.ConclusionsDistinct EV-driven signals from the microenvironment are capable to promote the TN malignant hemopoiesis and their further investigation paves the way toward novel therapeutic approaches for rare MF.
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