Comprehensive analysis of mesenchymal cells reveals a dysregulated TGF-β/WNT/HOXB7 axis in patients with myelofibrosis
Saravanan Ganesan, Sarah Awan-Toor, Fabien Guidez, Nabih Maslah, Rifkath Rahimy, Céline Aoun, Panhong Gou, Chloé Guiguen, Juliette Soret, Odonchimeg Ravdan, Valeria Bisio, Nicolas Dulphy, Camille Lobry, Marie-Hélène Schlageter, Michèle Souyri, Stéphane Giraudier, Jean-Jacques Kiladjian, Christine Chomienne, Bruno Cassinat
Saravanan Ganesan, Sarah Awan-Toor, Fabien Guidez, Nabih Maslah, Rifkath Rahimy, Céline Aoun, Panhong Gou, Chloé Guiguen, Juliette Soret, Odonchimeg Ravdan, Valeria Bisio, Nicolas Dulphy, Camille Lobry, Marie-Hélène Schlageter, Michèle Souyri, Stéphane Giraudier, Jean-Jacques Kiladjian, Christine Chomienne, Bruno Cassinat
View: Text | PDF
Research Article Hematology

Comprehensive analysis of mesenchymal cells reveals a dysregulated TGF-β/WNT/HOXB7 axis in patients with myelofibrosis

Abstract

Despite the advances in the understanding and treatment of myeloproliferative neoplasm (MPN), the disease remains incurable with the risk of evolution to acute myeloid leukemia or myelofibrosis (MF). Unfortunately, the evolution of the disease to MF remains poorly understood, impeding preventive and therapeutic options. Recent studies in solid tumor microenvironment and organ fibrosis have shed instrumental insights on their respective pathogenesis and drug resistance, yet such precise data are lacking in MPN. In this study, through a patient sample–driven transcriptomic and epigenetic description of the MF microenvironment landscape and cell-based analyses, we identify homeobox B7 (HOXB7) overexpression and more precisely a potentially novel TGF-β/WNT/HOXB7 pathway as associated to a pro-fibrotic and pro-osteoblastic biased differentiation of mesenchymal stromal cells (MSCs). Using gene-based and chemical inhibition of this pathway, we reversed the abnormal phenotype of MSCs from patients with MF, providing the MPN field a potentially novel target to prevent and manage evolution to MF.

Authors

Saravanan Ganesan, Sarah Awan-Toor, Fabien Guidez, Nabih Maslah, Rifkath Rahimy, Céline Aoun, Panhong Gou, Chloé Guiguen, Juliette Soret, Odonchimeg Ravdan, Valeria Bisio, Nicolas Dulphy, Camille Lobry, Marie-Hélène Schlageter, Michèle Souyri, Stéphane Giraudier, Jean-Jacques Kiladjian, Christine Chomienne, Bruno Cassinat

×

Figure 1

Expanded bone marrow MSCs from patients with MPN MF show an inflammatory profile and a biased osteoblast differentiation.

Options: View larger image (or click on image) Download as PowerPoint
Expanded bone marrow MSCs from patients with MPN MF show an inflammatory...
(A) Experimental workflow of this study (illustration created using BioRender). ATAC-Seq, assay for transposase-accessible chromatin using sequencing. (B) Representative phase contrast images of expanded bone marrow MSCs from either patients with MPN MF patients or healthy age-matched controls. The spindle shape morphology of control MSCs (left) shifts to a stellar morphology in F-MPNs (right). Pictures were taken on passage 2. Scale bar = 20× original magnification. (C) Immunophenotypic characterization of expanded F-MPNs and control MSCs. All markers were present but differentially expressed in F-MPNs compared with controls. P values (*<0.05, **<0.01, ***<0.001) were calculated using unpaired t test. (D) Cytokine secretion profile of expanded MSCs. Day 3 supernatants at passage 2 were analyzed using a multiplexed Luminex assay. F-MPNs show a significant increase in inflammatory cytokine levels. P values (*<0.05, **<0.01, ***<0.001) were calculated using unpaired t test. (E) Heatmap and unsupervised hierarchical clustering by sample and gene were performed using the 300 genes (RNA-Seq data) that had the largest coefficients of variation based on DESeq2 analysis. The data are based on samples from the F-MPNs (n = 7) and control (n = 4) MSC group. (F) Volcano plot showing the relationship between the P values and the log2 fold-change in normalized expression (DESeq2) between F-MPNs (n = 7) and control MSCs (n = 4). Genes found to be the most differentially expressed are shown in the plot by P value. (G) Gene set enrichment analysis (GSEA) of RNA-Seq data between F-MPNs (n = 7) and control MSCs (n = 4) demonstrates an enrichment of gene sets of the fibrosis pathway. (H) Bar graph showing the pathways differentially expressed in the RNA-Seq analysis from control MSCs versus F-MPNs identified by DAVID pathway analysis. (I) Bar graph showing upregulation of osteoblast-associated genes and downregulation of adipocyte-associated genes in RNA-Seq data of F-MPNs.