Osteogenesis depends on commissioning of a network of stem cell transcription factors that act as repressors of adipogenesis
Alexander Rauch, Anders K. Haakonsson, Jesper G. S. Madsen, Mette Larsen, Isabel Forss, Martin R. Madsen, Elvira L. Van Hauwaert, Christian Wiwie, Naja Z. Jespersen, Michaela Tencerova, Ronni Nielsen, Bjørk D. Larsen, Richard Röttger, Jan Baumbach, Camilla Scheele, Moustapha Kassem and Susanne Mandrup
Nat Genet. 2019 Apr;51(4):716-727. doi: 10.1038/s41588-019-0359-1.
Mesenchymal (stromal) stem cells (MSCs) constitute populations of mesodermal multipotent cells involved in tissue regeneration and homeostasis in many different organs. Here we performed comprehensive characterization of the transcriptional and epigenomic changes associated with osteoblast and adipocyte differentiation of human MSCs. We demonstrate that adipogenesis is driven by considerable remodeling of the chromatin landscape and de novo activation of enhancers, whereas osteogenesis involves activation of preestablished enhancers. Using machine learning algorithms for in silico modeling of transcriptional regulation, we identify a large and diverse transcriptional network of pro-osteogenic and antiadipogenic transcription factors. Intriguingly, binding motifs for these factors overlap with SNPs related to bone and fat formation in humans, and knockdown of single members of this network is sufficient to modulate differentiation in both directions, thus indicating that lineage determination is a delicate balance between the activities of many different transcription factors.
Chronic Kidney Disease Is Associated With Greater Bone Marrow Adiposity
Gina N Woods, Susan K Ewing, Sigurdur Sigurdsson, Deborah M Kado, Joachim H Ix,Trisha F Hue, Gudny Eiriksdottir, Kaipin Xu, Vilmundur Gudnason, Thomas F Lang, Eric Vittinghoff, Tamara B Harris, Clifford J Rosen, Xiaojuan Li, Ann V Schwartz. J Bone Miner Res.2018 Aug 3. doi: 10.1002/jbmr.3562
Bone marrow adiposity is associated with aging, osteoporosis, and reduced hematopoiesis, as well as anorexia nervosa, but little is known about the underlying mechanisms that affect marrow adiposity. Chronic kidney disease (CKD) may influence bone marrow adipose tissue (BMAT), possibly through loss of lean mass or higher circulating levels of sclerostin. To test these hypotheses, we investigated the cross‐sectional association between estimated glomerular filtration rate (eGFR) as a measure of kidney function and 1H‐MRS‐based measurement of vertebral BMAT (L1 to L4) in 475 older adults from the Age Gene/Environment Susceptibility (AGES)‐Reykjavik study. Mean BMAT was compared in those with eGFR >60 (n = 297) versus those with eGFR 45 to 60 (n = 120) or eGFR <45 (n = 58) using linear regression models. Participants had a mean age of 81.5 (SD 4.1) years, mean eGFR of 64.3 (SD 16.1) mL/min/1.734 cm2, mean BMAT of 54.5% (SD 8.5); 48.2% were women. In unadjusted and adjusted models (age, visit window, gender, diabetes and visceral adipose tissue), BMAT was higher in those with eGFR <45 (adjusted mean 58.5%; 95% CI, 56.2 to 60.7) compared with those with eGFR >60 (adjusted mean 53.8%; 95% CI, 52.8 to 54.8) (p = 0.0002). BMAT did not differ in those with eGFR 45 to 60 (adjusted mean 54.3%; 95% CI, 52.8 to 55.9) compared with those with eGFR >60 (p = 0.58). In a subgroup of participants with serum sclerostin available (n = 253), additional adjustment for sclerostin attenuated the difference in adjusted mean vertebral BMAT between those with eGFR <45 versus >60 from 3.7% (p = 0.04) to 2.4% (p = 0.20). CKD stage 3b or worse was associated with greater bone marrow adiposity; this association may be partially mediated by sclerostin.
April 2018 Specific Modulation of Vertebral Marrow Adipose Tissue by Physical Activity
Daniel L Belavy, Matthew J Quittner, Nicola D Ridgers, Adnan Shiekh, Timo Rantalainen,Guy Trudel. J Bone Miner Res. 2018 Apr;33(4):651-657. doi: 10.1002/jbmr.3357. Epub 2018 Jan 16.
Marrow adipose tissue (MAT) accumulation with normal aging impacts the bone, hemopoiesis, and metabolic pathways. We investigated whether exercise was associated with lower MAT, as measured by vertebral marrow fat fraction (VFF) on magnetic resonance imaging. A total of 101 healthy individuals (54 females) aged 25 to 35 years without spine or bone disease but with distinct exercise histories were studied. Long-distance runners (67 km/wk, n = 25) exhibited lower mean lumbar VFF (27.9% [8.6%] versus 33.5% [6.0%]; p = 0.0048) than non-sporting referents (n = 24). In habitual joggers (28 km/wk, n = 30), mean lumbar VFF was 31.3% (9.0%) (p = 0.22 versus referents) and 6.0 percentage points lower than referents at vertebrae T10 , T11 , and T12 (p ≤ 0.023). High-volume road cycling (275 km/wk, n = 22) did not impact VFF. 3D accelerations corresponding to faster walking, slow jogging, and high-impact activities correlated with lower VFF, whereas low-impact activities and sedentary time correlated with higher mean lumbar VFF (all p ≤ 0.05). Given an estimated adipose bone marrow conversion of 7% per decade of life, long distance runners, with 5.6 percentage points lower VFF, showed an estimated 8-year younger vertebral marrow adipose tissue phenotype. Regression analysis showed a 0.7 percentage point reduction in mean lumbar VFF with every 9.4 km/wk run (p = 0.002). This study presents the first evidence in humans or animals that specific volumes and types of exercise may influence the age-determined adipose marrow conversion and result in low MAT. These results identify a potentially modifiable risk factor for prevalent chronic conditions related to bone metabolism, hemopoietic production, and other metabolic functions with potential global health applications.
March 2018 Development of a 3D bone marrow adipose tissue model.
Heather Fairfield, Carolyne Falank, Mariah Farrell, CalvinVary, Joshua M. Boucher, HeatherDriscoll, Lucy Liaw, Clifford J. Rosen, Michaela R. Reagan. Bone. 2018 Jan 26. pii: S8756-3282(18)30023-1. doi: 10.1016/j.bone.2018.01.023.
Over the past twenty years, evidence has accumulated that biochemically and spatially defined networks of extracellular matrix, cellular components, and interactions dictate cellular differentiation, proliferation, and function in a variety of tissue and diseases. Modeling in vivo systems in vitro has been undeniably necessary, but when simplified 2D conditions rather than 3D in vitro models are used, the reliability and usefulness of the data derived from these models decreases. Thus, there is a pressing need to develop and validate reliable in vitro models to reproduce specific tissue-like structures and mimic functions and responses of cells in a more realistic manner for both drug screening/disease modeling and tissue regeneration applications. In adipose biology and cancer research, these models serve as physiologically relevant 3D platforms to bridge the divide between 2D cultures and in vivo models, bringing about more reliable and translationally useful data to accelerate benchtop to bedside research. Currently, no model has been developed for bone marrow adipose tissue (BMAT), a novel adipose depot that has previously been overlooked as “filler tissue” but has more recently been recognized as endocrine-signaling and systemically relevant. Herein we describe the development of the first 3D, BMAT model derived from either human or mouse bone marrow (BM) mesenchymal stromal cells (MSCs). We found that BMAT models can be stably cultured for at least 3 months in vitro, and that myeloma cells (5TGM1, OPM2 and MM1S cells) can be cultured on these for at least 2 weeks. Upon tumor cell co-culture, delipidation occurred in BMAT adipocytes, suggesting a bidirectional relationship between these two important cell types in the malignant BM niche. Overall, our studies suggest that 3D BMAT represents a “healthier,” more realistic tissue model that may be useful for elucidating the effects of MAT on tumor cells, and tumor cells on MAT, to identify novel therapeutic targets. In addition, proteomic characterization as well as microarray data (expression of >22,000 genes) coupled with KEGG pathway analysis and gene set expression analysis (GSEA) supported our development of less-inflammatory 3D BMAT compared to 2D culture. In sum, we developed the first 3D, tissue-engineered bone marrow adipose tissue model, which is a versatile, novel model that can be used to study numerous diseases and biological processes involved with the bone marrow.
February 2018 Characterization of the bone marrow adipocyte niche with three-dimensional electron microscopy
Hero Robles, Sung Jae Park, Matthew S. Joens, James A. J. Fitzpatrick, Clarissa S. Craft, Erica L. Scheller. Bone. 2018 Jan 27. pii: S8756-3282(18)30020-6. doi: 10.1016/j.bone.2018.01.020.
Unlike white and brown adipose tissues, the bone marrow adipocyte (BMA) exists in a microenvironment containing unique populations of hematopoietic and skeletal cells. To study this microenvironment at the sub-cellular level, we performed a three-dimensional analysis of the ultrastructure of the BMA niche with focused ion beam scanning electron microscopy (FIB-SEM). This revealed that BMAs display hallmarks of metabolically active cells including polarized lipid deposits, a dense mitochondrial network, and areas of endoplasmic reticulum. The distinct orientations of the triacylglycerol droplets suggest that fatty acids are taken up and/or released in three key areas – at the endothelial interface, into the hematopoietic milieu, and at the bone surface. Near the sinusoidal vasculature, endothelial cells send finger-like projections into the surface of the BMA which terminate near regions of lipid within the BMA cytoplasm. In some regions, perivascular cells encase the BMA with their flattened cellular projections, limiting contacts with other cells in the niche. In the hematopoietic milieu, BMAT adipocytes of the proximal tibia interact extensively with maturing cells of the myeloid/granulocyte lineage. Associations with erythroblast islands are also prominent. At the bone surface, the BMA extends organelle and lipid-rich cytoplasmic regions toward areas of active osteoblasts. This suggests that the BMA may serve to partition nutrient utilization between diverse cellular compartments, serving as an energy-rich hub of the stromal-reticular network. Lastly, though immuno-EM, we’ve identified a subset of bone marrow adipocytes that are innervated by the sympathetic nervous system, providing an additional mechanism for regulation of the BMA. In summary, this work reveals that the bone marrow adipocyte is a dynamic cell with substantial capacity for interactions with the diverse components of its surrounding microenvironment. These local interactions likely contribute to its unique regulation relative to peripheral adipose tissues.
January 2018 Acute myeloid leukaemia disrupts endogenous myelo-erythropoiesis by compromising the adipocyte bone marrow niche
Allison L. Boyd, Jennifer C. Reid, Kyle R. Salci, Lili Aslostovar, Yannick D. Benoit, Zoya, Mio Nakanishi, Deanna P. Porras, Mohammed Almakadi, Clinton J. V. Campbell, Michael F. , Catherine A. Ross, Ronan Foley, Brian Leber, David S. Allan, Mitchell Sabloff, Anargyros Xenocostas, Tony J. Collins and Mickie Bhatia. Nat Cell Biol. 2017 Nov;19(11):1336-1347. doi: 10.1038/ncb3625. Epub 2017 Oct 16
Acute myeloid leukaemia (AML) is distinguished by the generation of dysfunctional leukaemic blasts, and patients characteristically suffer from fatal infections and anaemia due to insufficient normal myelo-erythropoiesis. Direct physical crowding of bone marrow (BM) by accumulating leukaemic cells does not fully account for this haematopoietic failure. Here, analyses from AML patients were applied to both in vitro co-culture platforms and in vivo xenograft modelling, revealing that human AML disease specifically disrupts the adipocytic niche in BM. Leukaemic suppression of BM adipocytes led to imbalanced regulation of endogenous haematopoietic stem and progenitor cells, resulting in impaired myelo-erythroid maturation. In vivo administration of PPARγ agonists induced BM adipogenesis, which rescued healthy haematopoietic maturation while repressing leukaemic growth. Our study identifies a previously unappreciated axis between BM adipogenesis and normal myelo-erythroid maturation that is therapeutically accessible to improve symptoms of BM failure in AML via non-cell autonomous targeting of the niche.
December 2017 Clinical Implications of Bone Marrow Adiposity.
Veldhuis-Vlug AG, Rosen CJ J Intern Med. 2017 Dec 6. doi: 10.1111/joim.12718.
Marrow adipocytes, collectively termed marrow adipose tissue (MAT), reside in the bone marrow in close contact to bone cells and hematopoietic cells. Marrow adipocytes arise from the mesenchymal stem cell and share their origin with the osteoblastst. Shifts in the lineage allocation of the mesenchymal stromal cell could potentially explain the association between increased MAT and increased fracture risk in diseases such as postmenopausal osteoporosis, anorexia nervosa and diabetes. Functionally, marrow adipocytes secrete adipokines, such as adiponectin, and cytokines, such as RANK-ligand and stem cell factor. These mediators can influence both boneremodeling and hematopoiesis by promoting bone resorption and hematopoietic recovery following chemotherapy. In addition, marrowadipocytes can secrete free fatty acids, acting as a energy supply for bone and hematopoietic cells. However, this induced lipolysis is also used by neoplastic cells to promote survival and proliferation. Therefore, MAT could represent a new therapeutic target for multiple diseases from osteoporosis to leukemia, although the exaxt characteristics and role of the marrow adipocyte in health and diseases remains to be determined. This article is protected by copyright. All rights reserved.
November 2017 Bone Marrow Fat Changes After Gastric Bypass Surgery Are Associated With Loss of Bone Mass.
Kim TY, Schwartz AV, Li X, Xu K, Black DM, Petrenko DM, Stewart L, Rogers SJ, Posselt AM, Carter JT, Shoback DM, Schafer A J Bone Miner Res. 2017 Nov;32(11):2239-2247. doi: 10.1002/jbmr.3212. Epub 2017 Aug 9.
October 2017 Bone marrow adipocytes promote the regeneration of stem cells and haematopoiesis by secreting SCF.
Zhou BO, Yu H, Yue R, Zhao Z, Rios JJ, Naveiras O, Morrison SJ Nat Cell Biol. 2017 Aug;19(8):891-903. doi: 10.1038/ncb3570. Epub 2017 Jul 17.
Endothelial cells and leptin receptor+ (LepR+) stromal cells are critical sources of haematopoietic stem cell (HSC) niche factors, including stem cell factor (SCF), in bone marrow. After irradiation or chemotherapy, these cells are depleted while adipocytes become abundant. We discovered that bone marrow adipocytes synthesize SCF. They arise from Adipoq-Cre/ER+ progenitors, which represent ∼5% of LepR+ cells, and proliferate after irradiation. Scf deletion using Adipoq-Cre/ER inhibited haematopoietic regeneration after irradiation or 5-fluorouracil treatment, depleting HSCs and reducing mouse survival. Scf from LepR+ cells, but not endothelial, haematopoietic or osteoblastic cells, also promoted regeneration. In non-irradiated mice, Scf deletion using Adipoq-Cre/ER did not affect HSC frequency in long bones, which have few adipocytes, but depleted HSCs in tail vertebrae, which have abundant adipocytes. A-ZIP/F1 ‘fatless’ mice exhibited delayed haematopoietic regeneration in long bones but not in tail vertebrae, where adipocytes inhibited vascularization. Adipocytes are a niche component that promotes haematopoietic regeneration.
September 2017 Adipocyte Accumulation in the Bone Marrow during Obesity and Aging Impairs Stem Cell-Based Hematopoietic and Bone Regeneration.
Ambrosi TH, Scialdone A, Graja A, Gohlke S, Jank AM, Bocian C, Woelk L, Fan H, Logan DW, Schürmann A, Saraiva LR, Schulz TJ. Cell Stem Cell. 2017 Jun 1;20(6):771-784.e6. doi: 10.1016/j.stem.2017.02.009. Epub 2017 Mar 16.
Aging and obesity induce ectopic adipocyte accumulation in bone marrow cavities. This process is thought to impair osteogenic and hematopoietic regeneration. Here we specify the cellular identities of the adipogenic and osteogenic lineages of the bone. While aging impairs the osteogenic lineage, high-fat diet feeding activates expansion of the adipogenic lineage, an effect that is significantly enhanced in aged animals. We further describe a mesenchymal sub-population with stem cell-like characteristics that gives rise to both lineages and, at the same time, acts as a principal component of the hematopoietic niche by promoting competitive repopulation following lethal irradiation. Conversely, bone-resident cells committed to the adipocytic lineage inhibit hematopoiesis and bone healing, potentially by producing excessive amounts of Dipeptidyl peptidase-4, a protease that is a target of diabetes therapies. These studies delineate the molecular identity of the bone-resident adipocytic lineage, and they establish its involvement in age-dependent dysfunction of bone and hematopoietic regeneration.