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Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
Comparative Mutational Profiling of Hematopoietic Progenitor Cells and Circulating Endothelial Cells (CECs) in
Mirko Farina1,2, Simona Bernardi1,2, Nicola Polverelli1
1Unit of Blood Diseases and Bone Marrow Transplantation, Cell Therapies and Hematology Research Program, Department of Clinical and Experimental Sciences, University of Brescia, ASST Spedali Civili di Brescia, P.le Spedali Civili, 1, 25123 Brescia, Italy.
Abstract:
A role of endothelial cells (ECs) in Primary Myelofibrosis (PMF) was supposed since JAK2 mutation was found in endothelial precursor cells (EPCs) and in ECs captured by laser microdissection. By Cell Search method, the circulating endothelial cells (CECs) from 14 PMF patients and 5 healthy controls have been isolated and compared by NGS with CD34+Hematopoietic stem and progenitors cells (HSPCs) for panel of 54 myeloid-associated mutations. PMF patients had higher levels of CECs. No mutation was found in HSPCs and CECs from controls, while CECs from PMF patients presented several somatic mutations. 72% of evaluable patients shared at least one mutation between HSPCs and CECs. 2 patients shared the JAK2 mutation, together with ABL1, IDH1, TET2 and ASXL1, KMT2A, respectively. 6 out of 8 shared only NON MPN-driver mutations: TET2 and NOTCH1 in one case; individual paired mutations in TP53, KIT, SRSF2, NOTCH1 and WT1, in the other cases. In conclusion, 70% of PMF patients shared at least one mutation between HSPCs and CECs. These latter harbored several myeloid-associated mutations, besides JAK2V617F mutation. Our results support a primary involvement of EC in PMF and provide a new methodological approach for further studies exploring the role of the "neoplastic" vascular niche.
Insights
Endothelial cells (ECs) play a role in Primary Myelofibrosis (PMF). Circulating endothelial cells (CECs) in PMF patients harbor myeloid-associated mutations, suggesting EC involvement in the disease.
Area of Science:
- Hematology
- Oncology
- Molecular Biology
Background:
- Endothelial cells (ECs) are increasingly recognized for their role in hematological malignancies.
- Previous findings suggested a link between JAK2 mutations and endothelial cells in Primary Myelofibrosis (PMF).
Purpose of the Study:
- To investigate the presence and significance of somatic mutations in circulating endothelial cells (CECs) of PMF patients.
- To compare mutations in CECs with those in CD34+ hematopoietic stem and progenitor cells (HSPCs) in PMF.
- To explore the potential primary involvement of ECs in the pathogenesis of PMF.
Main Methods:
- Isolation of CECs from PMF patients and healthy controls using the Cell Search method.
- Next-generation sequencing (NGS) analysis of CECs and HSPCs for a panel of 54 myeloid-associated mutations.
- Comparison of mutation profiles between CECs and HSPCs within individual PMF patients.
Main Results:
- PMF patients exhibited significantly higher levels of CECs compared to healthy controls.
- CECs from PMF patients harbored various somatic mutations, including JAK2, ABL1, IDH1, TET2, ASXL1, KMT2A, TP53, KIT, NOTCH1, SRS2, and WT1.
- A high concordance rate (72%) of mutations was observed between CECs and HSPCs in PMF patients, with many non-driver mutations shared.
Conclusions:
- The findings support a primary role of endothelial cells in the development and progression of PMF.
- Shared mutations between CECs and HSPCs highlight the complex cellular interactions in PMF.
- This study introduces a novel methodological approach for investigating the contribution of the vascular niche in PMF.
