Extracellular vesicles derived from endometrial epithelial cells deliver exogenous miR-92b-3p to affect the function of embryonic trophoblast cells via targeting TSC1 and DKK3
Tóm tắt
Extracellular vesicles (EVs) could mediate embryo-maternal communication to affect embryo implantation by delivering biology information, including microRNA (miRNA), protein, lipid. Our previous research shows that miR-92b-3p was differentially expressed in EVs of uterine flushing fluids during the embryo implantation period. However, the role of miR-92b-3p from EVs in embryo implantation remains elusive. EVs were isolated from porcine endometrial epithelial cells (EECs) by ultracentrifugation. MiR-92b-3p mimics and EVs were used to regulate the expression of miR-92b-3p in porcine trophoblast cells (PTr2 cells). Cell proliferation, migration and adhesion analyses were used to observe the phenotype. RT-qPCR, western blot and dual-luciferase reporter assay were used to assess the targets of miR-92b-3p. In this study, EVs derived from porcine EECs were identified and could be taken up by PTr2 cells. We found that the EVs derived from EECs transfected with miR-92b-3p mimic (EVs-miR-92b-3p) significantly promoted the proliferation, migration and adhesion of PTr2 cells. We verified that Tuberous sclerosis complex subunit (TSC1) and Dickkopf 3 (DKK3) were the target genes of miR-92b-3p. Moreover, our study showed that miR-92b-3p plays a vital role in PTr2 cells via targeting TSC1 and DKK3. Furthermore, the 3'UTR vectors of TSC1 and DKK3 can rescue the effect of miR-92b-3p on PTr2 cells. Taken together, this study reveals a novel mechanism that EVs derived from porcine EECs treated with miR-92b-3p crosstalk with trophoblasts by targeting TSC1 and DKK3, leading to an enhanced ability for implantation.
Tài liệu tham khảo
Tayade C, Fang Y, Croy BA. A review of gene expression in porcine endometrial lymphocytes, endothelium and trophoblast during pregnancy success and failure. J Reprod Dev. 2007;53:455–63.
Yelich JV, Pomp D, Geisert RD. Detection of transcripts for retinoic acid receptors, retinol-binding protein, and transforming growth factors during rapid trophoblastic elongation in the porcine conceptus. Biol Reprod. 1997;57:286–94.
Achache H, Revel A. Endometrial receptivity markers, the journey to successful embryo implantation. Hum Reprod Update. 2006;12:731–46.
Vigano P, Mangioni S, Pompei F, Chiodo I. Maternal-conceptus cross talk--a review. Placenta. 2003;24(Suppl B):S56-61.
Bidarimath M, Khalaj K, Kridli RT, Kan FW, Koti M, Tayade C. Extracellular vesicle mediated intercellular communication at the porcine maternal-fetal interface: A new paradigm for conceptus-endometrial cross-talk. Sci Rep. 2017;7:40476.
Kurian NK, Modi D. Extracellular vesicle mediated embryo-endometrial cross talk during implantation and in pregnancy. J Assist Reprod Genet. 2019;36:189–98.
Thery C, Zitvogel L, Amigorena S. Exosomes: composition, biogenesis and function. Nat Rev Immunol. 2002;2:569–79.
Turturici G, Tinnirello R, Sconzo G, Geraci F. Extracellular membrane vesicles as a mechanism of cell-to-cell communication: advantages and disadvantages. Am J Physiol Cell Physiol. 2014;306:C621–33.
Hugel B, Martinez MC, Kunzelmann C, Freyssinet JM. Membrane microparticles: two sides of the coin. Physiology (Bethesda). 2005;20:22–7.
Burns G, Brooks K, Wildung M, Navakanitworakul R, Christenson LK, Spencer TE. Extracellular vesicles in luminal fluid of the ovine uterus. PLoS ONE. 2014;9:e90913.
Krawczynski K, Bauersachs S, Reliszko ZP, Graf A, Kaczmarek MM. Expression of microRNAs and isomiRs in the porcine endometrium: implications for gene regulation at the maternal-conceptus interface. BMC Genomics. 2015;16:906.
Kusama K, Nakamura K, Bai R, Nagaoka K, Sakurai T, Imakawa K. Intrauterine exosomes are required for bovine conceptus implantation. Biochem Biophys Res Commun. 2018;495:1370–5.
Lv C, Yu WX, Wang Y, Yi DJ, Zeng MH, Xiao HM. MiR-21 in extracellular vesicles contributes to the growth of fertilized eggs and embryo development in mice. Biosci Rep. 2018;38:BSR20180036.
Vilella F, Moreno-Moya JM, Balaguer N, Grasso A, Herrero M, Martinez S, Marcilla A, Simon C. Hsa-miR-30d, secreted by the human endometrium, is taken up by the pre-implantation embryo and might modify its transcriptome. Development. 2015;142:3210–21.
Yáñez-Mó M, Siljander PRM, Andreu Z, Zavec AB, Borràs FE, Buzas EI, Buzas K, Casal E, Cappello F, Carvalho J, et al. Biological properties of extracellular vesicles and their physiological functions. Journal of extracellular vesicles. 2015;4:27066.
Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol. 2013;200:373–83.
Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, Antoniou A, Arab T, Archer F, Atkin-Smith GK, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2018;7:1535750.
Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007;9:654–9.
Tan Q, Shi S, Liang J, Cao D, Wang S, Wang Z. Endometrial cell-derived small extracellular vesicle miR-100-5p promotes functions of trophoblast during embryo implantation. Mol Ther Nucleic Acids. 2021;23:217–31.
Hua R, Wang Y, Lian W, Li W, Xi Y, Xue S, Kang T, Lei M. Small RNA-seq analysis of extracellular vesicles from porcine uterine flushing fluids during peri-implantation. Gene. 2021;766:145117.
Guo S, Chen J, Chen F, Zeng Q, Liu WL, Zhang G. Exosomes derived from Fusobacterium nucleatum-infected colorectal cancer cells facilitate tumour metastasis by selectively carrying miR-1246/92b-3p/27a-3p and CXCL16. Gut. 2021;70:1507–19.
Gao X, Ran N, Dong X, Zuo B, Yang R, Zhou Q, Moulton HM, Seow Y, Yin H. Anchor peptide captures, targets, and loads exosomes of diverse origins for diagnostics and therapy. Sci Transl Med. 2018;10:0195.
Thery C, Amigorena S, Raposo G, Clayton A: Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol. 2006;Chapter 3:Unit 3.22.1-29.
Hua R, Zhang X, Li W, Lian W, Liu Q, Gao D, Wang Y, Lei M. Ssc-miR-21–5p regulates endometrial epithelial cell proliferation, apoptosis and migration via the PDCD4/AKT pathway. J cell sci. 2020;133:jcs248898.
Jaeger LA, Spiegel AK, Ing NH, Johnson GA, Bazer FW, Burghardt RC. Functional effects of transforming growth factor beta on adhesive properties of porcine trophectoderm. Endocrinology. 2005;146:3933–42.
Fu Y, Li L, Fang X, Li B, Zhao W, Zhou L, Ren S. Investigation of Eph-ephrin A1 in the regulation of embryo implantation in sows. Reprod Domest Anim. 2018;53:1563–74.
Jiang X, Xue S, Kang T, Liu H, Ren H, Hua R, Ni D, Lei M. Annexin A8 (ANXA8) regulates proliferation of porcine endometrial cells via Akt signalling pathway. Reprod Domest Anim. 2019;54:3–10.
Sun X, Ruan YC, Guo J, Chen H, Tsang LL, Zhang X, Jiang X, Chan HC. Regulation of miR-101/miR-199a-3p by the epithelial sodium channel during embryo implantation: involvement of CREB phosphorylation. Reproduction. 2014;148:559–68.
Wang Y, Hua R, Xue S, Li W, Wu L, Kang T, Lei M. mRNA/lncRNA expression patterns and the function of fibrinogen-like protein 2 in Meishan pig endometrium during the preimplantation phases. Mol Reprod Dev. 2019;86:354–69.
Hu Q, Zang X, Ding Y, Gu T, Shi J, Li Z, Cai G, Liu D, Wu Z, Hong L. Porcine uterine luminal fluid-derived extracellular vesicles improve conceptus-endometrial interaction during implantation. Theriogenology. 2021;178:8–17.
Zhou C, Cai G, Meng F, Xu Z, He Y, Hu Q, Zheng E, Huang S, Xu Z, Gu T, et al. Deep-Sequencing Identification of MicroRNA Biomarkers in Serum Exosomes for Early Pig Pregnancy. Front Genet. 2020;11:536.
Nguyen HP, Simpson RJ, Salamonsen LA, Greening DW. Extracellular Vesicles in the Intrauterine Environment: Challenges and Potential Functions. Biol Reprod. 2016;95:109.
Greening DW, Nguyen HPT, Elgass K, Simpson RJ, Salamonsen LA. Human Endometrial Exosomes Contain Hormone-Specific Cargo Modulating Trophoblast Adhesive Capacity: Insights into Endometrial-Embryo Interactions. Biol Reprod. 2016;94:38.
Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, Ofrecio JM, Wollam J, Hernandez-Carretero A, Fu W, et al. Adipose Tissue Macrophage-Derived Exosomal miRNAs Can Modulate In Vivo and In Vitro Insulin Sensitivity. Cell. 2017;171(372–384):e312.
Ali A, Bouma GJ, Anthony RV, Winger QA. The Role of LIN28--ARID3B Pathway in Placental Development. Int J Mol Sci. 2020;21:3637.
Wang J, Armant DR. Integrin-mediated adhesion and signaling during blastocyst implantation. Cells Tissues Organs. 2002;172:190–201.
Zhang Z, Zhuang L, Lin C-P. Roles of MicroRNAs in Establishing and Modulating Stem Cell Potential. International journal of molecular sciences. 2019;20:3643.
Lee J, Heo J, Kang H. miR-92b-3p-TSC1 axis is critical for mTOR signaling-mediated vascular smooth muscle cell proliferation induced by hypoxia. Cell Death Differ. 2019;26:1782–95.
Li T, Liu X, Gong X. E Q, Zhang X, Zhang X: microRNA 92b–3p regulates primordial follicle assembly by targeting TSC1 in neonatal mouse ovaries. Cell Cycle. 2019;18:824–33.
Wang LP, Geng JN, Sun B, Sun CB, Shi Y, Yu XY. MiR-92b-3p is Induced by Advanced Glycation End Products and Involved in the Pathogenesis of Diabetic Nephropathy. Evid Based Complement Alternat Med. 2020;2020:6050874.
Daikoku T, Yoshie M, Xie H, Sun X, Cha J, Ellenson LH, Dey SK. Conditional deletion of Tsc1 in the female reproductive tract impedes normal oviductal and uterine function by enhancing mTORC1 signaling in mice. Mol Hum Reprod. 2013;19:463–72.
Aghajanova L, Shen S, Rojas AM, Fisher SJ, Irwin JC, Giudice LC. Comparative transcriptome analysis of human trophectoderm and embryonic stem cell-derived trophoblasts reveal key participants in early implantation. Biology of reproduction. 2012;86:1–21.