Trophoblast-derived hyaluronan promotes the regulatory phenotype of decidual macrophages

in Reproduction
Correspondence should be addressed to R Zhu: summerrui2006@aliyun.com or M Du: dmrlq1973@sina.cn or H Li: hongziszivf@163.com

*(S Wang and F Sun contributed equally to this work)

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There is delicate crosstalk between fetus-derived trophoblasts (Tros) and maternal cells during normal pregnancy. Dysfunctions in interaction are highly linked to some pregnancy complications, such as recurrent spontaneous abortion (RSA), pre-eclampsia and fetal growth restriction. Hyaluronan (HA), the most abundant component of extracellular matrix, has been reported to act as both a pro- and an anti-inflammatory molecule. Previously, we reported that HA promotes the invasion and proliferation of Tros by activating PI3K/Akt and MAPK/ERK1/2 signaling pathways. While lower HA secretion by Tros was observed during miscarriages than that during normal pregnancies, in the present study, we further confirmed that higher secretion of HA by Tros could induce M2 polarization of macrophages at the maternal–fetal interface by interacting with CD44 and activating the downstream PI3K/Akt-STAT-3/STAT-6 signaling pathways. Furthermore, HA could restore the production of IL-10 and other normal pregnancy markers by decidual macrophages (dMφs) from RSA. These findings underline the important roles of HA in regulating the function of dMφs and maintaining a normal pregnancy.

 

    Society for Reproduction and Fertility

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    Effects of HA on dMφs during early pregnancy. (A) Quantitation of flow cytometric analysis of the expression of MHC II and co-stimulatory molecules by dMφs stimulated with HMW-HA (HHA, 50 or 100 μg/mL) or MMW-HA (MHA, 50 or 100 μg/mL) or LMW-HA (LHA, 50 or 100 μg/mL). (B, C and D) Quantitation of flow cytometric analysis of the expression of CD206, CD163 cytokines and chemokines by dMφs stimulated with the indicated concentrations of HMW-HA, MMW-HA or LMW-HA. Data are represented as the mean ± standard error of the mean (s.e.m.) for three to five separate experiments. n = 12 dMφs in the first trimester of normal pregnancies. *P < 0.05, **P < 0.01, ***P < 0.001.

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    HMW-HA induces M2 polarization through CD44. (A) Quantitation of flow cytometric analysis of the expression of CD44 on DICs. (B) Flow cytometric analysis of the expression of CD44 on dMφs. The flow cytometric plot is from one representative experiment among three separate experiments. (C) Quantitation of flow cytometric analysis of M1/M2 associated markers on dMφs stimulated with HMW-HA (HHA, 100 μg/mL) in the presence or absence of anti-CD44 mAbs. Data are represented as the mean ± s.e.m. for three separate experiments. n = 9 dMφs in the first trimester of normal pregnancies. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the group Ctrl. # P < 0.5, ## P < 0.01, ### P < 0.001, compared with the group HHA.

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    PI3K/Akt-STAT-3/STAT-6 pathways are involved in the M2 polarization. (A) Quantitation of flow cytometric analysis of M1/M2 associated markers on dMφs stimulated with HMW-HA (HHA, 100 μg/mL) in the presence or absence of the indicated inhibitors of signal transduction. (B) dMφs were serum treated with 100 μg/mL HMW-HA at a different time. A Western blot was used to analyze the total levels and phosphorylation levels of Akt. HMW-HA induced Akt phosphorylation in dMφs in a time-dependent manner. The image is representative of four individual experiments. (C) Flow cytometric analysis (right) and quantitation (left) of STAT-3 and STAT-6 phosphorylation in dMφs stimulated with HMW-HA (HHA, 100 μg/mL) in the presence or absence of the PI3K signaling pathway inhibitor LY294002. The flow cytometric plot is from one representative experiment. Data are represented as the mean ± s.e.m. for three separate experiments, n = 9. *P < 0.05, **P < 0.01, ***P < 0.001, compared with group Ctrl. ## P < 0.01, ### P < 0.001, compared with group HHA.

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    Tros instruct dMφs to an M2 phenotype via HMW-HA. Quantitation of flow cytometric analysis of M1/M2 associated markers on dMφs cultured with TCM in the presence or absence of pep-1 and Cpep with or without pretreatment of neutralizing anti-CD44 (20 µg/mL) antibody. Data are represented as the mean ± s.e.m. for three separate experiments, n = 12. *P < 0.05, **P < 0.01, ***P < 0.001, compared with group Ctrl. # P < 0.5, ## P < 0.01, ### P < 0.001, compared with group TCM + Cpep.

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    Dysregulation of dMφs correlates with human miscarriages. (A) Flow cytometric analysis (right) and quantitation (left) of MHC II and co-stimulatory molecules on dMφs from normal pregnancies (n = 12) and RSA (n = 19). (B) Flow cytometric analysis (right) and quantitation (left) of cytokine production by dMφs from normal pregnancies and RSA. (C) Flow cytometric analysis (right) and quantitation (left) of pattern recognition receptors on dMφs from normal pregnancy and RSA. The flow cytometric plot is from one representative experiment among six separate experiments. (D) Quantitation of flow cytometric analysis of the surface molecule expression and cytokine production by dMφs isolated from patients with RSA (n = 9) stimulated with HMW-HA (HHA, 100 μg/mL) from three separate experiments. Data are represented as the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001.

  • View in gallery

    A schematic diagram of Tros-derived HA induced M2 polarization at the maternal–fetal interface. Tros secrete HA, which in turn promotes the invasion and proliferation of human Tros. Tros-derived HMW-HA instructs dMφs to transform into immunosuppressive M2 through interacting with CD44 and activating the downstream PI3K/Akt-STAT-3/STAT-6 signaling pathways. While in RSA, the secretion of HA by Tros was decreased, and the M2 bias was disrupted at the maternal–fetal interface, which was harmful to the maintenance of normal pregnancy.

References

  • AkgulYWordRAEnsignLMYamaguchiYLydonJHanesJMahendrooM 2014 Hyaluronan in cervical epithelia protects against infection-mediated preterm birth. Journalof Clinical Investigation 124 54815489. (https://doi.org/10.1172/JCI78765)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ArckPCHecherK 2013 Fetomaternal immune cross-talk and its consequences for maternal and offspring’s health. Nature Medicine 19 548556. (https://doi.org/10.1038/nm.3160)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • AriyoshiWOkinagaTKnudsonCBKnudsonWNishiharaT 2014 High molecular weight hyaluronic acid regulates osteoclast formation by inhibiting receptor activator of NF-kappaB ligand through Rho kinase. Osteoarthritis Cartilage 22 111120. (https://doi.org/10.1016/j.joca.2013.10.013)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • CareASDienerKRJasperMJBrownHMIngmanWVRobertsonSA 2013 Macrophages regulate corpus luteum development during embryo implantation in mice. Journal of Clinical Investigation 123 34723487. (https://doi.org/10.1172/JCI60561)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ChanmeeTOntongPItanoN 2016 Hyaluronan: a modulator of the tumor microenvironment. Cancer Letters 375 2030. (https://doi.org/10.1016/j.canlet.2016.02.031)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • DelFCOteroKGomez-GarciaLGonzalez-LeonMCSoler-RangerLFuentes-PriorPEscollPBaosRCavedaLGarciaF 2005 Tumor cells deactivate human monocytes by up-regulating IL-1 receptor associated kinase-M expression via CD44 and TLR4. Journal of Immunology 174 30323040. (https://doi.org/10.4049/jimmunol.174.5.3032)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • DickerKTGurskiLAPradhan-BhattSWittRLFarach-CarsonMCJiaX 2014 Hyaluronan: a simple polysaccharide with diverse biological functions. Acta Biomaterialia 10 15581570. (https://doi.org/10.1016/j.actbio.2013.12.019)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • EgashiraMHirotaYShimizu-HirotaRSaito-FujitaTHaraguchiHMatsumotoLMatsuoMHiraokaTTanakaTAkaedaS 2017 F4/80+ macrophages contribute to clearance of senescent cells in the mouse postpartum uterus. Endocrinology 158 23442353. (https://doi.org/10.1210/en.2016-1886)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HeldinPKarousouEBernertBPorschHNishitsukaKSkandalisSS 2008 Importance of hyaluronan–CD44 interactions in inflammation and tumorigenesis. Connective Tissue Research 49 215218. (https://doi.org/10.1080/03008200802143323)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HighleyCBPrestwichGDBurdickJA 2016 Recent advances in hyaluronic acid hydrogels for biomedical applications. Current Opinion in Biotechnology 40 3540. (https://doi.org/10.1016/j.copbio.2016.02.008)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • IshiiMWenHCorsaCALiuTCoelhoALAllenRMCarsonWTCavassaniKALiXLukacsNW 2009 Epigenetic regulation of the alternatively activated macrophage phenotype. Blood 114 32443254. (https://doi.org/10.1182/blood-2009-04-217620)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • JaiswalMKMallersTMLarsenBKwak-KimJChaouatGGilman-SachsABeamanKD 2012 V-ATPase upregulation during early pregnancy: a possible link to establishment of an inflammatory response during preimplantation period of pregnancy. Reproduction 143 713725. (https://doi.org/10.1530/REP-12-0036)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • JiangDLiangJFanJYuSChenSLuoYPrestwichGDMascarenhasMMGargHGQuinnDA 2005 Regulation of lung injury and repair by Toll-like receptors and hyaluronan. Nature Medicine 11 11731179. (https://doi.org/10.1038/nm1315)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • JiangDLiangJNoblePW 2011 Hyaluronan as an immune regulator in human diseases. Physiological Reviews 91 221264. (https://doi.org/10.1152/physrev.00052.2009)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • LashGEPitmanHMorganHLInnesBAAgwuCNBulmerJN 2016 Decidual macrophages: key regulators of vascular remodeling in human pregnancy. Journal of Leukocyte Biology 100 315325. (https://doi.org/10.1189/jlb.1A0815-351R)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • MillsCDKincaidKAltJMHeilmanMJHillAM 2000 M-1/M-2 macrophages and the Th1/Th2 paradigm. Journal of Immunology 164 61666173. (https://doi.org/10.4049/jimmunol.164.12.6166)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • MurrayPJAllenJEBiswasSKFisherEAGilroyDWGoerdtSGordonSHamiltonJAIvashkivLBLawrenceT 2014 Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41 1420. (https://doi.org/10.1016/j.immuni.2014.06.008)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • PetroffMGSedlmayrPAzzolaDHuntJS 2002 Decidual macrophages are potentially susceptible to inhibition by class Ia and class Ib HLA molecules. Journal of Reproductive Immunology 56 317. (https://doi.org/10.1016/S0165-0378(02)00024-4)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • PowellJDHortonMR 2005 Threat matrix: low-molecular-weight hyaluronan (HA) as a danger signal. Immunologic Research 31 207218. (https://doi.org/10.1385/IR:31:3:207)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • RayahinJEBuhrmanJSZhangYKohTJGemeinhartRA 2015 High and low molecular weight hyaluronic acid differentially influence macrophage activation. ACS Biomaterials Science and Engineering 1 481493. (https://doi.org/10.1021/acsbiomaterials.5b00181)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ShimadaSEbinaYIijimaNDeguchiMYamadaH 2018 Decidual CD68(+) HLA-DR(+) CD163(−) M1 macrophages increase in miscarriages with normal fetal chromosome. American Journal of Reprodective Immunology 79 e12791. (https://doi.org/10.1111/aji.12791)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • SicaABronteV 2007 Altered macrophage differentiation and immune dysfunction in tumor development. Journal of Clinical Investigation 117 11551166. (https://doi.org/10.1172/JCI31422)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • SmithPWalshCMManganNEFallonRESayersJRMcKenzieANFallonPG 2004 Schistosoma mansoni worms induce anergy of T cells via selective up-regulation of programmed death ligand 1 on macrophages. Journal of Immunology 173 12401248. (https://doi.org/10.4049/jimmunol.173.2.1240)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • SternRKoganGJedrzejasMJSoltesL 2007 The many ways to cleave hyaluronan. Biotechnology Advances 25 537557. (https://doi.org/10.1016/j.biotechadv.2007.07.001)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Svensson-ArvelundJErnerudhJ 2015 The role of macrophages in promoting and maintaining homeostasis at the fetal–maternal interface. American Journal of Reproductive Immunology 74 100109. (https://doi.org/10.1111/aji.12357)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • TangMXHuXHLiuZZKwak-KimJLiaoAH 2015 What are the roles of macrophages and monocytes in human pregnancy? Journal of Reproductive Immunology 112 7380. (https://doi.org/10.1016/j.jri.2015.08.001)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • TerrazasLIMonteroDTerrazasCAReyesJLRodriguez-SosaM 2005 Role of the programmed death-1 pathway in the suppressive activity of alternatively activated macrophages in experimental cysticercosis. International Journal for Parasitology 35 13491358. (https://doi.org/10.1016/j.ijpara.2005.06.003)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • TiainenSTumeliusRRillaKHamalainenKTammiMTammiRKosmaVMOikariSAuvinenP 2015 High numbers of macrophages, especially M2-like (CD163-positive), correlate with hyaluronan accumulation and poor outcome in breast cancer. Histopathology 66 873883. (https://doi.org/10.1111/his.12607)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • TsaoFYWuMYChangYLWuCTHoHN 2018 M1 macrophages decrease in the deciduae from normal pregnancies but not from spontaneous abortions or unexplained recurrent spontaneous abortions. Journal of the Formosan Medical Association 117 204211. (https://doi.org/10.1016/j.jfma.2017.03.011)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • WangSCLiYHPiaoHLHongXWZhangDXuYYTaoYWangYYuanMMLiDJ 2015 PD-1 and Tim-3 pathways are associated with regulatory CD8+ T-cell function in decidua and maintenance of normal pregnancy. Cell Death and Disease 6 e1738. (https://doi.org/10.1038/cddis.2015.112)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • WangSZhuXXuYZhangDLiYTaoYPiaoHLiDDuM 2016 Programmed cell death-1 (PD-1) and T-cell immunoglobulin mucin-3 (Tim-3) regulate CD4+ T cells to induce Type 2 helper T cell (Th2) bias at the maternal–fetal interface. Human Reproduction 31 700711. (https://doi.org/10.1093/humrep/dew019)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • WelchJSEscoubet-LozachLSykesDBLiddiardKGreavesDRGlassCK 2002 TH2 cytokines and allergic challenge induce Ym1 expression in macrophages by a STAT6-dependent mechanism. Journal of Biological Chemistry 277 4282142829. (https://doi.org/10.1074/jbc.M205873200)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • XuYRomeroRMillerDKadamLMialTNPlazyoOGarcia-FloresVHassanSSXuZTarcaAL 2016 An M1-like macrophage polarization in decidual tissue during spontaneous preterm labor that is attenuated by rosiglitazone treatment. Journal of Immunology 196 24762491. (https://doi.org/10.4049/jimmunol.1502055)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ZhangGGuoLYangCLiuYHeYDuYWangWGaoF 2016 A novel role of breast cancer-derived hyaluronan on inducement of M2-like tumor-associated macrophages formation. OncoImmunology 5 e1172154. (https://doi.org/10.1080/2162402X.2016.1172154)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • ZhangYHHeMWangYLiaoAH 2017 Modulators of the balance between M1 and M2 macrophages during pregnancy. Frontiers in Immunology 8 120. (https://doi.org/10.3389/fimmu.2017.00120)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • ZhouJMiaoHLiXHuYSunHHouY 2017 Curcumin inhibits placental inflammation to ameliorate LPS-induced adverse pregnancy outcomes in mice via upregulation of phosphorylated Akt. Inflammation Research 66 177185. (https://doi.org/10.1007/s00011-016-1004-4)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ZhuRHuangYHTaoYWangSCSunCPiaoHLWangXQDuMRLiDJ 2013a Hyaluronan up-regulates growth and invasion of trophoblasts in an autocrine manner via PI3K/AKT and MAPK/ERK1/2 pathways in early human pregnancy. Placenta 34 784791. (https://doi.org/10.1016/j.placenta.2013.05.009)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • ZhuRWangSCSunCTaoYPiaoHLWangXQDuMRLiDJ 2013b Hyaluronan–CD44 interaction promotes growth of decidual stromal cells in human first-trimester pregnancy. PLoS ONE 8 e74812. (https://doi.org/10.1371/journal.pone.0074812)

    • Crossref
    • Search Google Scholar
    • Export Citation

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