AML-induced osteogenic differentiation in mesenchymal stromal cells supports leukemia growth.

Genotypic and phenotypic alterations in the bone marrow (BM) microenvironment, in particular in osteoprogenitor cells, have been shown to support leukemogenesis. However, it is unclear how leukemia cells alter the BM microenvironment to create a hospitable niche. Here, we report that acute myeloid leukemia (AML) cells, but not normal CD34+ or CD33+ cells, induce osteogenic differentiation in mesenchymal stromal cells (MSCs). In addition, AML cells inhibited adipogenic differentiation of MSCs. Mechanistic studies identified that AML-derived BMPs activate Smad1/5 signaling to induce osteogenic differentiation in MSCs. Gene expression array analysis revealed that AML cells induce connective tissue growth factor (CTGF) expression in BM-MSCs irrespective of AML type. Overexpression of CTGF in a transgenic mouse model greatly enhanced leukemia engraftment in vivo. Together, our data suggest that AML cells induce a preosteoblast-rich niche in the BM that in turn enhances AML expansion.

[1]  Gang Huang,et al.  Leukaemogenic effects of Ptpn11 activating mutations in the stem cell microenvironment , 2016, Nature.

[2]  H. Mewes,et al.  Stroma-Derived Connective Tissue Growth Factor Maintains Cell Cycle Progression and Repopulation Activity of Hematopoietic Stem Cells In Vitro , 2015, Stem cell reports.

[3]  P. Sikorski,et al.  Osteogenic Differentiation of Human Mesenchymal Stem Cells in Mineralized Alginate Matrices , 2015, PloS one.

[4]  J. Mao,et al.  CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model. , 2010, The Journal of clinical investigation.

[5]  J. Teruya-Feldstein,et al.  Inhibition of leukemia cell engraftment and disease progression in mice by osteoblasts. , 2014, Blood.

[6]  M. Guindani,et al.  A role for BMP-induced homeobox gene MIXL1 in acute myelogenous leukemia and identification of type I BMP receptor as a potential target for therapy , 2014, Oncotarget.

[7]  S. Armstrong,et al.  Acute myelogenous leukemia-induced sympathetic neuropathy promotes malignancy in an altered hematopoietic stem cell niche. , 2014, Cell stem cell.

[8]  T. Shibahara,et al.  A Novel Strategy for Enrichment and Isolation of Osteoprogenitor Cells from Induced Pluripotent Stem Cells Based on Surface Marker Combination , 2014, PloS one.

[9]  R. Rabadán,et al.  Leukemogenesis Induced by an Activating β-catenin mutation in Osteoblasts , 2014, Nature.

[10]  M. Konopleva,et al.  Targeting connective tissue growth factor (CTGF) in acute lymphoblastic leukemia preclinical models: anti-CTGF monoclonal antibody attenuates leukemia growth , 2014, Annals of Hematology.

[11]  Ralph Müller,et al.  A comparative analysis of water-soluble and blood-pool contrast agents for in vivo vascular imaging with micro-CT. , 2013, Academic radiology.

[12]  D. Strunk,et al.  Connective tissue growth factor regulates adipocyte differentiation of mesenchymal stromal cells and facilitates leukemia bone marrow engraftment. , 2013, Blood.

[13]  A. Bullock,et al.  Development of an ALK2-biased BMP type I receptor kinase inhibitor. , 2013, ACS chemical biology.

[14]  D. Abraham,et al.  Connective tissue growth factor causes EMT-like cell fate changes in vivo and in vitro , 2013, Journal of Cell Science.

[15]  M. Konopleva,et al.  The Bone Marrow Microenvironment as Niche Retreats for Hematopoietic and Leukemic Stem Cells , 2013, Advances in hematology.

[16]  W. Tawackoli,et al.  BMP-6 is more efficient in bone formation than BMP-2 when overexpressed in mesenchymal stem cells , 2012, Gene Therapy.

[17]  M. L. Le Beau,et al.  BMP meets AML: induction of BMP signaling by a novel fusion gene promotes pediatric acute leukemia. , 2012, Cancer cell.

[18]  A. Reinisch,et al.  Human extramedullary bone marrow in mice: a novel in vivo model of genetically controlled hematopoietic microenvironment. , 2012, Blood.

[19]  P. Becker,et al.  Dependence of Acute Myeloid Leukemia on Adhesion within the Bone Marrow Microenvironment , 2012, TheScientificWorldJournal.

[20]  B. Cohen,et al.  Detection of Intranasally Delivered Bone Marrow-Derived Mesenchymal Stromal Cells in the Lesioned Mouse Brain: A Cautionary Report , 2011, Stem cells international.

[21]  M. Konopleva,et al.  Leukemia stem cells and microenvironment: biology and therapeutic targeting. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  Alex G. Lambi,et al.  The role of connective tissue growth factor (CTGF/CCN2) in skeletogenesis. , 2011, Critical reviews in eukaryotic gene expression.

[23]  Charles P. Lin,et al.  Bone progenitor dysfunction induces myelodysplasia and secondary leukemia , 2010, Nature.

[24]  D. Strunk,et al.  Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures. , 2009, Journal of visualized experiments : JoVE.

[25]  A. Miyawaki,et al.  Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow , 2009, The Journal of experimental medicine.

[26]  T. He,et al.  BMP‐9‐induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/β‐catenin signalling , 2009, Journal of cellular and molecular medicine.

[27]  J. Downing,et al.  Mouse models of human AML accurately predict chemotherapy response. , 2009, Genes & development.

[28]  L. Kanz,et al.  Isolation of functionally distinct mesenchymal stem cell subsets using antibodies against CD56, CD271, and mesenchymal stem cell antigen-1 , 2009, Haematologica.

[29]  Angela C. Colmone,et al.  Leukemic Cells Create Bone Marrow Niches That Disrupt the Behavior of Normal Hematopoietic Progenitor Cells , 2008, Science.

[30]  Sean J. Morrison,et al.  Stem Cells and Niches: Mechanisms That Promote Stem Cell Maintenance throughout Life , 2008, Cell.

[31]  Lothar Just,et al.  Human placenta and bone marrow derived MSC cultured in serum-free, b-FGF-containing medium express cell surface frizzled-9 and SSEA-4 and give rise to multilineage differentiation. , 2007, Differentiation; research in biological diversity.

[32]  H. Gundacker,et al.  Connective tissue growth factor (CTGF) expression and outcome in adult patients with acute lymphoblastic leukemia. , 2007, Blood.

[33]  M. Long,et al.  Osteogenic differentiation of human mesenchymal stem cells is regulated by bone morphogenetic protein‐6 , 2006, Journal of cellular biochemistry.

[34]  C. Morsczeck.  Gene Expression of runx2, Osterix, c-fos, DLX-3, DLX-5, and MSX-2 in Dental Follicle Cells during Osteogenic Differentiation In Vitro , 2006, Calcified Tissue International.

[35]  A. Montag,et al.  Connective Tissue Growth Factor (CTGF) Is Regulated by Wnt and Bone Morphogenetic Proteins Signaling in Osteoblast Differentiation of Mesenchymal Stem Cells* , 2004, Journal of Biological Chemistry.

[36]  J. Szatkowski,et al.  Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery , 2004, Gene Therapy.

[37]  D. Prockop,et al.  An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. , 2004, Analytical biochemistry.

[38]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[39]  D. Scadden,et al.  Osteoblastic cells regulate the haematopoietic stem cell niche , 2003, Nature.

[40]  R. Kusec,et al.  Expression of bone morphogenetic proteins in acute promyelocytic leukemia before and after combined all trans-retinoic acid and cytotoxic treatment. , 2003, Leukemia research.

[41]  Jie Xu,et al.  Expression of connective tissue growth factor in bone: Its role in osteoblast proliferation and differentiation in vitro and bone formation in vivo , 2003, Journal of cellular physiology.

[42]  M. Konopleva,et al.  Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins , 2002, Leukemia.

[43]  Robert E. Lenkinski,et al.  In vivo near-infrared fluorescence imaging of osteoblastic activity , 2001, Nature Biotechnology.

[44]  B. Hogan,et al.  Bone morphogenetic proteins: multifunctional regulators of vertebrate development. , 1996, Genes & development.