Periostin supports hematopoietic progenitor cells and niche-dependent myeloblastoma cells in vitro

Abstract

The expression of extracellular matrix protein periostin (POSTN) was attenuated in Med1−/− mouse embryonic fibroblasts (MEFs), which exhibited a decreased capability to support hematopoietic progenitor cells (HPCs) in vitro. When bone marrow (BM) cells were cocultured with mitomycin C-treated Med1+/+ MEFs, or OP-9 or MS-5 BM stromal cells, in the presence of anti-POSTN antibody, the growth of BM cells and number of long-term culture-initiating cells (LTC-ICs) were attenuated. When BM cells were cocultured with Med1−/− MEFs in the presence of recombinant POSTN, the growth of BM cells and the number of LTC-ICs were restored. Moreover, antibody-mediated blockage of stromal cells-derived POSTN markedly reduced the growth and cobblestone formation, a leukemic stem cell feature, of stromal cell-dependent MB-1 myeloblastoma cells. POSTN was expressed both in BM cells and variably in different BM stromal cells. Expression in the latter cells was increased by physical interaction with hematopoietic cells. The receptor for POSTN, integrin αvβ3, was expressed abundantly in BM stromal cells. The addition of recombinant POSTN to BM stromal cells induced intracellular signaling downstream of integrin αvβ3. These results suggest that stromal cell POSTN supports both normal HPCs and leukemia-initiating cells in vitro, at least in part, indirectly by acting on stromal cells in an autocrine or paracrine manner.

Periostin function in communication with extracellular matrices

“…Interestingly, there are several recent reports of periostin function in stem cells. Periostin has been shown to maintain hematopoietic stem cells (HSC) in a quiescent state through its interaction with integrin-αv of HSC in the bone marrow niche (Khurana et al 2016); and another recent report showed that periostin secreted from stromal fibroblastic cells supports not only normal hematopoietic precursor cells but also leukemia-initiating cells (Tanaka et al 2016). It has also been reported that periostin associates with a matricellular protein, CCN3 (also termed NOV), and that a functional relationship was observed in the periodontal ligament (Djokic et al 2013).…”

Matrix Gla protein maintains normal and malignant hematopoietic progenitor cells by interacting with bone morphogenetic protein-4

“…Mouse BM-derived primary mesenchymal stem/stromal cells (MSCs; Gibco), and Med1 þ/þ and Med1 À/À mouse embryonic fibroblasts (MEFs) that harbored a p53 inactivation [10], were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) [10]. The MB-1 niche-dependent myeloblastoma cells [26,27] were maintained by coculturing with mitomycin C (MMC)-treated OP-9 cells and used by coculturing with MMC-treated either MS-5 or OP-9 cells [12]. In some experiments, 0.5 μM LDN-193189 (Selleck) or 10 μM warfarin (Wako) was added to the coculture [24].…”

“…BM hematopoietic cells were prepared from femora and tibiae of C57BL6 mice [12]. These cells (5 Â 10 5 cells/well) were added to 24-well plates in which MMC-treated MS-5 or OP-9 cells, or MEFs, were pre-seeded, and cultured in Methocult M5300 (STEMCELL Technologies) in the absence or presence of 0.5 mg/ml anti-mouse (m) MGP goat polyclonal antibody (sc-32820; Santa Cruz Biotechnology) or normal goat IgG (R&D Systems) [10].…”

“…For long-term culture-initiating cells (LTC-ICs), BM hematopoietic cells cocultured with MMC-treated MS-5 or OP-9 stromal cells for 4 weeks were placed in complete methylcellulose medium (Methocult M3434; STEMCELL Technologies) for 2 weeks, and colonies were counted [10,12].…”

“…Among the circa 31 Mediator subunits, MED1 is a specific coactivator for activators including nuclear receptors [7] and is crucial for transcription of genes involved in hematopoietic niche function [10]. We previously identified dozens of genes whose expressions were significantly attenuated in Med1 À/À mesenchymal stromal cells [10], and reported that osteopontin, fibroblast growth factor 7 (FGF7) and periostin independently supported hematopoietic stem or progenitor cells (HSPCs) in vitro [10,11,12].…”