Self-repopulating recipient bone marrow resident macrophages promote long-term hematopoietic stem cell engraftment

Kaur, Simranpreet; Raggatt, Liza J.; Millard, Susan; Wu, Andy C.K.; Batoon, Lena; Jacobsen, Rebecca; Winkler, Ingrid G.; MacDonald, Kelli P. A.; Perkins, Andrew C.; Hume, David; Pettit, Allison R.

doi:10.1182/blood-2018-01-829663

Cited by 73 publications

(76 citation statements)

References 54 publications

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“…For liver and brain Percoll density gradient centrifugation was used to isolate the nonparenchymal fraction, as previously described (4,5). Cell isolation from BM, spleen and lymph node (LN) was carried out as described (54). X cells were stained for flow cytometry analysis and 1-2 x 106 cells were acquired for analysis.…”

Section: Tissue Collection For Imaging and Disaggregation For Flow Cymentioning

confidence: 99%

A transgenic line that reports CSF1R protein expression provides a definitive marker for the mouse mononuclear phagocyte system

Grabert

Sehgal

Irvine

et al. 2020

Preprint

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The proliferation, differentiation and survival of cells of the mononuclear phagocyte system (MPS, progenitors, monocytes, macrophages and classical dendritic cells) is controlled by signals from the macrophage colony-stimulating factor receptor (CSF1R). Cells of the MPS lineage have been identified using numerous surface markers and transgenic reporters but none is both universal and lineage-restricted. Here we report the development and characterization of a novel CSF1R reporter mouse. A Fusion Red (FRed) cassette was inserted in-frame with the C-terminus of CSF1R, separated by a T2A-cleavable linker. The insertion had no effect of CSF1R expression or function. CSF1R-FRed was expressed in monocytes and macrophages and absent from granulocytes and lymphocytes. In bone marrow, CSF1R-FRed was absent in lineage-negative hematopoietic stem cells (HSC), arguing against a direct role for CSF1R in myeloid lineage commitment. It was highly-expressed in marrow monocytes and common myeloid progenitors (CMP) but significantly lower in granulocyte-macrophage progenitors (GMP). In sections of bone marrow, CSF1R-FRed was also detected in osteoclasts, CD169+ resident macrophages and, consistent with previous mRNA analysis, in megakaryocytes. In lymphoid tissues, CSF1R-FRed highlighted diverse MPS populations including classical dendritic cells. Whole mount imaging of non-lymphoid tissues in mice with combined CSF1R-FRed/Csf1r-EGFP confirmed the restriction of CSF1R expression to MPS cells. The two markers highlight the remarkable abundance and regular distribution of tissue MPS cells including novel macrophage populations within tendon and skeletal muscle and underlying the mesothelial/serosal/capsular surfaces of every major organ. The CSF1R-FRed mouse provides a novel reporter with exquisite specificity for cells of the MPS.

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Section: Tissue Collection For Imaging and Disaggregation For Flow Cymentioning

confidence: 99%

A transgenic line that reports CSF1R protein expression provides a definitive marker for the mouse mononuclear phagocyte system

Grabert

Sehgal

Irvine

et al. 2020

Preprint

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“…Myeloid lineage 7,28 , HSPC 7,29 , B cell subsets 30,31 and red blood cell maturation 29,32,33 phenotyping were performed as detailed in Supplementary Table 1. Cell acquisition was performed on Beckman Coulter's CyAn™ ADP Analyser (Beckman Coulter, USA), Cytoflex Analyser (Beckman Coulter, USA) or BD LSRFortessa™ X-20 (BD Biosciences, USA) for panels specified in Supplementary Table 1.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…CD169 is expressed by HSC 5,7 and erythroid 39,40 niche macrophages in the BM. Given both the sustained increase in spleen size and shift in splenic red pulp macrophage phenotype to express CD169 5,7 , we assessed whether CSF1-Fc treatment induced extramedullary haematopoiesis and/or impacted HSPC populations in BM, spleen and liver at 7 and 14 days post treatment.…”

Section: Csf1-fc Treatment Transiently Increased Hspc Populations Inmentioning

confidence: 99%

“…To determine whether the increased splenic HSC pool observed at 14 days post-CSF1-Fc treatment was an indirect consequence of compensatory extramedullary haematopoiesis, myeloid progenitors 7,28 , lymphoid progenitors 29,30,47 , erythroblasts and reticulocytes 32,33 were assessed in BM, spleen and liver at day 14 post initial CSF1-Fc or saline treatment. No effect on common myeloid progenitors (CMP) number was observed at this time point in either BM or spleen, whereas they were increased in the liver (Fig.…”

Section: Csf1-fc Treatment Transiently Increased Hspc Populations Inmentioning

confidence: 99%

“…HSC reside in specific locations called niches in the bone marrow (BM) and BM resident macrophages are an integral component of HSC niches. Macrophage depletion in vivo is sufficient to drive mobilisation of HSC to blood [4][5][6] and prevent successful re-establishment of the HSC niche after total body irradiation 7 . Granulocyte colony stimulating factor (G-CSF) was one of the first growth factors used to mobilise HSC 8 and remains the main compound to elicit HSPC mobilisation in donors and patients 9 .…”

Section: Introductionmentioning

confidence: 99%

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Stable colony stimulating factor 1 fusion protein treatment increases HSC pool and enhances their mobilisation in mice

Kaur

Sehgal

et al. 2020

Preprint

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Prior chemotherapy and/or underlying morbidity commonly leads to poor mobilisation of hematopoietic stem cells (HSC) for transplantation in cancer patients. Increasing the number of available HSC prior to mobilisation is a potential strategy to overcome this deficiency. Resident bone marrow (BM) macrophages are essential for maintenance of niches that support HSC and enable engraftment in transplant recipients. Here we examined potential of donor treatment with colony stimulating factor-1 (CSF1) to modify the BM niche and expand the potential HSC pool for autologous transplantation. We administrated CSF1 Fc fusion protein (CSF1-Fc) to naive C57Bl/6 mice and assessed the impacts on HSC number and function and overall haematopoiesis. Outcomes were assessed by in situ imaging and ex vivo flow cytometry with functional validation by colony formation and competitive transplantation assay. CSF1-Fc treatment caused a transient expansion of monocyte-macrophage cells within BM and spleen at the expense of BM B lymphopoiesis and hematopoietic stem and progenitor cell (HSPC) homeostasis. During the recovery phase after cessation of CSF1-Fc treatment, normalisation of haematopoiesis was accompanied by an increase in the total available HSPC pool. In the spleen, increased HSC was associated with expression of the BM niche marker CD169 in red pulp macrophages. Pre-treatment with CSF1-Fc increased the number and reconstitution potential of HSPC in blood following a HSC mobilising regimen of granulocyte colony stimulating factor (G-CSF) treatment. These results indicate that CSF1-Fc conditioning could represent a therapeutic strategy to overcome poor HSC mobilisation and subsequently improve autologous or heterologous HSC transplantation outcomes.

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Neonatal Osteomacs and Bone Marrow Macrophages Differ in Phenotypic Marker Expression and Function

Mohamad

Gunawan

Blosser

et al. 2020

Journal of Bone and Mineral Research

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Osteomacs (OM) are specialized bone‐resident macrophages that are a component of the hematopoietic niche and support bone formation. Also located in the niche are a second subset of macrophages, namely bone marrow–derived macrophages (BM Mφ). We previously reported that a subpopulation of OM co‐express both CD166 and CSF1R, the receptor for macrophage colony‐stimulating factor (MCSF), and that OM form more bone‐resorbing osteoclasts than BM Mφ. Reported here are single‐cell quantitative RT‐PCR (qRT‐PCR), mass cytometry (CyTOF), and marker‐specific functional studies that further identify differences between OM and BM Mφ from neonatal C57Bl/6 mice. Although OM express higher levels of CSF1R and MCSF, they do not respond to MCSF‐induced proliferation, in contrast to BM Mφ. Moreover, receptor activator of NF‐κB ligand (RANKL), without the addition of MCSF, was sufficient to induce osteoclast formation in OM but not BM Mφ cultures. OM express higher levels of CD166 than BM Mφ, and we found that osteoclast formation by CD166−/− OM was reduced compared with wild‐type (WT) OM, whereas CD166−/− BM Mφ showed enhanced osteoclast formation. CD110/c‐Mpl, the receptor for thrombopoietin (TPO), was also higher in OM, but TPO did not alter OM‐derived osteoclast formation, whereas TPO stimulated BM Mφ osteoclast formation. CyTOF analyses demonstrated OM uniquely co‐express CD86 and CD206, markers of M1 and M2 polarized macrophages, respectively. OM performed equivalent phagocytosis in response to LPS or IL‐4/IL‐10, which induce polarization to M1 and M2 subtypes, respectively, whereas BM Mφ were less competent at phagocytosis when polarized to the M2 subtype. Moreover, in contrast to BM Mφ, LPS treatment of OM led to the upregulation of CD80, an M1 marker, as well as IL‐10 and IL‐6, known anti‐inflammatory cytokines. Overall, these data reveal that OM and BM Mφ are distinct subgroups of macrophages, whose phenotypic and functional differences in proliferation, phagocytosis, and osteoclast formation may contribute physiological specificity during health and disease. © 2021 American Society for Bone and Mineral Research (ASBMR).

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Self-repopulating recipient bone marrow resident macrophages promote long-term hematopoietic stem cell engraftment

Cited by 73 publications

References 54 publications

A transgenic line that reports CSF1R protein expression provides a definitive marker for the mouse mononuclear phagocyte system

A transgenic line that reports CSF1R protein expression provides a definitive marker for the mouse mononuclear phagocyte system

Stable colony stimulating factor 1 fusion protein treatment increases HSC pool and enhances their mobilisation in mice

Neonatal Osteomacs and Bone Marrow Macrophages Differ in Phenotypic Marker Expression and Function

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