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Journal of Experimental & Clinical Cancer Research

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Open Access 01-12-2021 | Research

Distinct profile of CD34⁺ cells and plasma-derived extracellular vesicles from triple-negative patients with Myelofibrosis reveals potential markers of aggressive disease

Authors: Dorian Forte, Martina Barone, Cristina Morsiani, Giorgia Simonetti, Francesco Fabbri, Samantha Bruno, Erika Bandini, Daria Sollazzo, Salvatore Collura, Maria Chiara Deregibus, Giuseppe Auteri, Emanuela Ottaviani, Nicola Vianelli, Giovanni Camussi, Claudio Franceschi, Miriam Capri, Francesca Palandri, Michele Cavo, Lucia Catani

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2021

Abstract

Background

Myelofibrosis (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.

Methods

Peripheral 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)-1β, Tumor Necrosis Factor (TNF)-α, IL6) or after co-culture with EVs from MF patients/HD.

Results

The 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.

Conclusions

Distinct 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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Title: Distinct profile of CD34+ cells and plasma-derived extracellular vesicles from triple-negative patients with Myelofibrosis reveals potential markers of aggressive disease
Authors: Dorian Forte
Martina Barone
Cristina Morsiani
Giorgia Simonetti
Francesco Fabbri
Samantha Bruno
Erika Bandini
Daria Sollazzo
Salvatore Collura
Maria Chiara Deregibus
Giuseppe Auteri
Emanuela Ottaviani
Nicola Vianelli
Giovanni Camussi
Claudio Franceschi
Miriam Capri
Francesca Palandri
Michele Cavo
Lucia Catani
Publication date: 01-12-2021
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2021
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-020-01776-8

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