Activated fibronectin-secretory phenotype of mesenchymal stromal cells in pre-fibrotic myeloproliferative neoplasms
© Schneider et al.; licensee BioMed Central Ltd. 2014
Received: 13 November 2014
Accepted: 29 November 2014
Published: 14 December 2014
We characterized bone marrow stromal cells (BMSC) from patients with pre-fibrotic myeloproliferative neoplasms (MPN). MPN-BMSC showed decreased capacity to stimulate the proliferation of colony-forming units of normal hematopoietic stem and progenitor cells and displayed increased matrix remodelling (in particular fibronectin deposition) compared to control BMSC. This finding was confirmed in pre-fibrotic MPN bone marrow biopsies in a tissue microarray (n = 34), which stained positive for fibronectin in the absence of reticulin as a standard myelofibrosis marker. Fibronectin expression correlated significantly with reduced haemoglobin levels in MPN-patients (p = 0.007; R2 = 0.42). Our data show significant cell-intrinsic alterations in MPN-MSC and suggest that Fibronectin expression might be applicable as a biomarker for the identification of early myelofibrotic transformation in reticulin-negative MPN.
To characterize the hematopoiesis-supporting capacity of MSC, secreted cytokines and their biological activity were tested. ET-MSC secreted significantly lower levels of G-CSF and IL-7 compared to controls, indicating a defect in the hematopoiesis-supporting capacity (Figure 1c). We therefore evaluated the biological activity of secreted cytokines in myeloid colony-forming unit (CFU) assays using human cord blood (CB) CD34+-hematopoietic stem and progenitor cells (HSPCs) . Myeloid CFU-activity significantly decreased when supernatants from PV and CML were used (Figure 1d). These findings indicate that MPN-BMSCs lose their capacity to produce and release functional myeloid differentiation-supporting cytokines.
To validate these findings, we systematically analyzed CD271 and fibronectin in tissue microarrays including ET (n = 13), PV (n = 11), CML (n = 14), PMF (n = 11) and non-MPN controls (n = 17), (Figure 2c). CD271 and fibronectin were evaluated applying a grading scheme similar to the established guidelines for grading myelofibrosis – 0 = no fibrosis and 3 = severe myelofibrosis . Although the majority of MPN cases were classified as myelofibrosis grade 0–1 in reticulin-staining, myelofibrotic transformation was graded as ≥ 1 by Fibronectin-staining in 69% of ET (vs. 7.7% Reticulin-representing one post ET-MF patient), 63% of PV (vs. 0% Reticulin) and 100% CML (vs. 0% Reticulin) cases and by CD271 staining in 54% of ET, 51% PV and 64% CML cases (Figure 2d). In line with the observation of a pre-fibrotic marrow, fibronectin and CD271 expression grade correlated significantly with decreased haemoglobin levels in reticulin-negative biopsies (Figure 2e).
In conclusion, our data reveal an intrinsic defect of MSC in pre-fibrotic MPN resulting in decreased hematopoiesis-supporting capacities and increased ECM remodelling. Our data suggest that fibronectin up-regulation/distribution detects early myelofibrotic changes in the BM of MPN patients and implies clinical and prognostic application in different myeloproliferative (Ph− and Ph+) neoplasms.
This work was supported by the START-program (to RKS and PZ) and the Interdisciplinary Center for Clinical Research (IZKF grant O1-6) both within the Faculty of Medicine RWTH Aachen University. R.K.S was supported by the German Research Foundation (DFG1188/3-1). We thank the “Immunohistochemistry and confocal microscopy facility”, a core facility of the Interdisciplinary Center for Clinical Research (IZKF) Aachen within the Faculty of Medicine at RWTH Aachen University. The institutional review board of the medical faculty of the RWTH Aachen approved the project (EK173/06).
- Kramann R, Couson SK, Neuss S, Kunter U, Bovi M, Bornemann J, Knuchel R, Jahnen-Dechent W, Floege J, Schneider RK: Exposure to uremic serum induces a procalcific phenotype in human mesenchymal stem cells. Arterioscler Thromb Vasc Biol. 2011, 31 (9): e45-54. 10.1161/ATVBAHA.111.228601.View ArticlePubMedGoogle Scholar
- Schneider RK, Puellen A, Kramann R, Raupach K, Bornemann J, Knuechel R, Perez-Bouza A, Neuss S: The osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. Biomaterials. 2010, 31 (3): 467-480. 10.1016/j.biomaterials.2009.09.059.View ArticlePubMedGoogle Scholar
- Manz MG, Miyamoto T, Akashi K, Weissman IL: Prospective isolation of human clonogenic common myeloid progenitors. Proc Natl Acad Sci U S A. 2002, 99 (18): 11872-11877. 10.1073/pnas.172384399.PubMed CentralView ArticlePubMedGoogle Scholar
- Tormin A, Li O, Brune JC, Walsh S, Schutz B, Ehinger M, Ditzel N, Kassem M, Scheding S: CD146 expression on primary nonhematopoietic bone marrow stem cells is correlated with in situ localization. Blood. 2011, 117 (19): 5067-5077. 10.1182/blood-2010-08-304287.PubMed CentralView ArticlePubMedGoogle Scholar
- Lo Celso C, Fleming HE, Wu JW, Zhao CX, Miake-Lye S, Fujisaki J, Cote D, Rowe DW, Lin CP, Scadden DT: Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche. Nature. 2009, 457 (7225): 92-96. 10.1038/nature07434.PubMed CentralView ArticlePubMedGoogle Scholar
- Thiele J, Kvasnicka HM, Facchetti F, Franco V, van der Walt J, Orazi A: European consensus on grading bone marrow fibrosis and assessment of cellularity. Haematologica. 2005, 90 (8): 1128-1132.PubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.