- Short report
- Open Access
Microbiota regulates bone marrow mesenchymal stem cell lineage differentiation and immunomodulation
- E Xiao†1, 2,
- Linhai He†1, 2,
- Qiong Wu4, 5,
- Junxiang Li4, 5,
- Yang He1, 2,
- Lu Zhao1, 2,
- Shuo Chen1, 2,
- Jingang An1, 2,
- Yansong Liu6,
- Chider Chen3Email author and
- Yi Zhang1, 2Email author
© The Author(s). 2017
- Received: 2 July 2017
- Accepted: 12 September 2017
- Published: 29 September 2017
Health is dependent on the homeostasis of both inner and external microenvironments. The microbiota as the external microenvironment plays a critical role in regulation of several organ systems in mammals. However, it is unclear whether the microbiota regulates homeostasis of the skeletal system and bone marrow mesenchymal stem cells (BMMSCs). Here, using a well-established germ-free (GF) mouse model, we show that the microbiota significantly alters the stemness of BMMSCs in comparison to specific-pathogen-free (SPF)-derived BMMSCs. Colonization of GF mice with SPF microbiota (conventionalized (ConvD)) normalizes the proliferation and differentiation abilities of BMMSCs. On the other hand, normal microbiota is required to maintain immunomodulatory properties of BMMSCs through induction of activated T-cell apoptosis and cytokine secretion. GF-derived BMMSCs lose the capacity to ameliorate disease phenotypes in dextran sulfate sodium-induced experimental colitis mice. Mechanistically, single-cell RNA-sequencing analysis shows that ConvD BMMSCs have a similar gene expression pattern to SPF-derived BMMSCs, which have a distinct gene distribution from GF-derived BMMSCs.
The mammal is inhabited by a vast number of bacteria, archaea, viruses, and eukaryotes. This microorganism coexistence with their hosts is referred to as the microbiota. It is reported that the human microbiota contains as many as 1014 bacterial cells, a number 10 times greater than the number of human cells . The microbiota colonizes on the host mammal after they are exposed to the external environment. More than a billion years of mammalian–microbial coevolution has led to interdependency, resulting in a critical role of the microbiota in hematopoiesis , immune system development , neurologic signaling , host metabolism , and bone mass remodeling .
Bone marrow mesenchymal stem cells (BMMSCs), a kind of adult stromal cell in bone marrow, both contribute to the bone turnover  and form the unique bone marrow niche with hematopoietic stem cells . BMMSCs show promising therapeutic potential based on their multipotent differentiation potential and immunomodulatory capacity [9, 10]. However, whether these mesenchymal stem cells are “born with” these fantastic capacities or are educated by the microbiota was still not known. Thus, in this study we aimed to elucidate the effect of the microbiota on the multipotent differentiation and immunomodulatory abilities of BMMSCs.
BMMSCs from germ-free mice exhibited higher colony forming ability and proliferation rate
Microbiota increases adipogenesis but decreases osteogenesis of BMMSCs
Collectively, these findings indicated that the microbiota inhibited BMMSC proliferation and adipogenesis but increased osteogenesis and in-vivo regenerative abilities. Interestingly, the microbiota increases adipogenic differentiation, which is consistent with reduced obesity in germ-free mice . Probiotic and prebiotic treatments are able to increase bone mass and BMD, indicating that intestinal microbiota may impact bone metabolism and health maintenance [13–16]. These findings connect gut microbiota and skeletal remodeling, which prompts us to investigate the impact of the microbiota on BMMSCs and bone tissue regeneration. They also suggest that physiological regulation of the osteoblastic/adipogenic lineage switch in the bone compartment involves the microbiota.
BMMSCs from GF mice were deficient in immunomodulation
The role of the microbiota in regulating immune cell polarization and human diseases is receiving increasing attention [22, 23]. BMMSCs reside in the skeleton to maintain osteoblastic lineage cell function and serve as a niche for hematopoietic stem cells, which involves several physiological regulations and interplays with the immune system. Our data indicated that in the unique niche of both mesenchymal stem cells and hematopoietic stem cells, the microbiota also regulated the BMMSCs, which may be related to the maturation or apoptosis of hematopoietic cell lines.
Single-cell RNA-sequencing analysis identified three pathway categories regulated by microbiota
The microbiota is involved in the regulation of multiple host metabolic pathways, which activates immune-inflammatory axes and signaling pathways . As the bone compartment is unlikely to directly contact with microbes, it is easy to envision that the microbiome could influence BMMSCs through regulating metabolic pathways. Our single-cell RNA-sequencing data further confirm that several major metabolic pathways are significantly different between GF and SPF/ConvD BMMSCs, implying metabolism may connect the microbiota and BMMSCs to maintain bone homeostasis. Besides, our results show that HIF-1 signaling may be the major regulator in BMMSC immunomodulation, since HIF-1 has been reported to crosstalk with inflammatory transcription factor NFκB and regulates release of cytokines and chemokines to control immune response [28, 29]. In summary, this is the first study to link the microbiota with BMMSC function, and single-cell RNA-sequencing analysis further provides detailed pathway prediction to connect the microbiota to regulating BMMSCs and bone metabolism.
In conclusion, we have revealed that the microbiota alters the differentiation potential and enhances the immunomodulation capacity of BMMSCs. This study provides a new point of view on how BMMSCs gain their therapeutic function.
The authors thank Ting Zhang from Department of Orthodontics, Peking University School and Hospital of Stomatology for her help in mouse BMMSC culture.
This work was supported by grants from National Natural Science Foundation of China (No. 81500819 and No. 81371117) and National Institute of Dental and Craniofacial Research, National Institutes of Health, Department of Health and Human Services (K99E025915 to CC).
Availability of data and materials
All data generated or analyzed during this study are included in this published article.
All methods and materials used in this study are listed in Additional file 3.
EX contributed to conception, design, and data acquisition. LHH contributed to data acquisition and analysis. QW and JXL contributed to single-cell sequencing and data analysis. HY, ZL, CS, and JGA contributed to the data collection, analysis, and interpretation. YSL contributed to raising mice and mice sample harvest. CC contributed to conception, design, and data acquisition and drafted the manuscript. YZ contributed to conception and design, and critically revised the manuscript. All authors read and approved the final manuscript.
All animal experiments were approved by The Ethics Committee of the Peking University Health Science Center (LA2016149).
Consent for publication
The authors declare that they have no competing interests.
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