Restricted accessResearch articleFirst published online 2020-08
Exosomes Derived from Human Placental Mesenchymal Stromal Cells Carrying AntagomiR-4450 Alleviate Intervertebral Disc Degeneration Through Upregulation of ZNF121
Exosomes derived from mesenchymal stromal cells (MSCs) have emerged as novel drug and gene delivery tools. Current study aimed to elucidate the potential therapeutic role of human placental MSC (hPLMSC)-derived exosomes carrying AntagomiR-4450 (EXO-AntagomiR-4450) in intervertebral disc degeneration (IDD) progression. Initially, the differentially expressed miRNAs related to IDD were identified by microarray analysis, which provided data predicting the interaction between microRNA-4450 (miR-4450) and zinc finger protein-121 (ZNF121) in IDD. Next, miR-4450 and ZNF121 were elevated or silenced to determine their effects on the damage of nucleus pulposus cells (NPCs) treated with tumor necrosis factor α (TNF-α). The therapeutic effects of EXO-AntagomiR-4450 on NPCs were verified both in vitro and in vivo (15-week-old C57BL/6 male mice); especially gait analysis and fluorescent molecular tomography were used in live mice with IDD. Our results revealed that miR-4450 was highly expressed, while ZNF121 was poorly expressed in IDD patients and NPCs treated with TNF-α. Furthermore, miR-4450 was identified to specifically target ZNF121. In addition, the inhibition of miR-4450 exerted an alleviatory effect on the inflammation, apoptosis, and damage of the NPCs by upregulating ZNF121 (all P < 0.05). Moreover, EXO-AntagomiR-4450 retarded damage of NPCs in vitro, alleviated IDD damage, and ameliorated gait abnormality in vivo (all P < 0.05). hPLMSC-derived exosomes could be a feasible nanovehicle to deliver inhibitory oligonucleotides like AntagomiR-4450 in IDD.
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References
1.
VosT, BarberRM, BellB, Bertozzi-VillaA and BiryukovS. (2015). Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet, 386:743–800.
2.
RobertsS, EvansH, TrivediJ and MenageJ. (2006). Histology and pathology of the human intervertebral disc. J Bone Joint Surg Am 88, (Suppl. 2)::10–14.
3.
PengB, ChenJ, KuangZ, LiD, PangX and ZhangX. (2009). Expression and role of connective tissue growth factor in painful disc fibrosis and degeneration. Spine (Phila Pa 1976), 34:E178–E182.
4.
YangSH, WuCC, ShihTT, SunYH and LinFH. (2008). In vitro study on interaction between human nucleus pulposus cells and mesenchymal stem cells through paracrine stimulation. Spine (Phila Pa 1976), 33:1951–1957.
5.
SemenovOV, KoestenbauerS, RiegelM, ZechN, ZimmermannR, ZischAH and MalekA. (2010). Multipotent mesenchymal stem cells from human placenta: critical parameters for isolation and maintenance of stemness after isolation. Am J Obstet Gynecol, 202:193.e1–193.e13.
6.
MiaoZ, JinJ, ChenL, ZhuJ, HuangW, ZhaoJ, QianH and ZhangX. (2006). Isolation of mesenchymal stem cells from human placenta: comparison with human bone marrow mesenchymal stem cells. Cell Biol Int, 30:681–687.
ShahabipourF, BaratiN, JohnstonTP, DerosaG, MaffioliP and SahebkarA. (2017). Exosomes: nanoparticulate tools for RNA interference and drug delivery. J Cell Physiol, 232:1660–1668.
10.
AngerF, CamaraM, EllingerE, GermerCT, SchlegelN, OttoC and KleinI. (2019). Human mesenchymal stromal cell-derived extracellular vesicles improve liver regeneration after ischemia reperfusion injury in mice. Stem Cells Dev, 28:1451–1462.
11.
ShabbirA, CoxA, Rodriguez-MenocalL, SalgadoM and Van BadiavasE. (2015). Mesenchymal stem cell exosomes induce proliferation and migration of normal and chronic wound fibroblasts, and enhance angiogenesis in vitro. Stem Cells Dev, 24:1635–1647.
LuK, LiHY, YangK, WuJL, CaiXW, ZhouY and LiCQ. (2017). Exosomes as potential alternatives to stem cell therapy for intervertebral disc degeneration: in-vitro study on exosomes in interaction of nucleus pulposus cells and bone marrow mesenchymal stem cells. Stem Cell Res Ther, 8:108.
14.
WangC, WangW-J, YanY-G, XiangY-X, ZhangJ, TangZ-H and JiangZ-S. (2015). MicroRNAs: new players in intervertebral disc degeneration. Clin Chim Acta, 450:333–341.
15.
MarcusME and LeonardJN. (2013). FedExosomes: engineering therapeutic biological nanoparticles that truly deliver. Pharmaceuticals (Basel), 6:659–680.
16.
LiuX, CheL, XieY-K, HuQ-J, MaC-J, PeiY-J, WuZ-G, LiuZ-H, FanL-Y and WangH-Q. (2015). Noncoding RNAs in human intervertebral disc degeneration: an integrated microarray study. Genomics Data, 5:80.
17.
SmythG. (2004). Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol, 3:Article3.
18.
PfirrmannCW, MetzdorfA, ZanettiM, HodlerJ and BoosN. (2001). Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976), 26:1873–1878.
19.
WangF, WuXT, ZhuangSY, WangYT, HongX, ZhuL and BaoJP. (2011). Ex vivo observation of human nucleus pulposus chondrocytes isolated from degenerated intervertebral discs. Asian Spine J, 5:73–81.
20.
JohnsonZI, SchoepflinZR, ChoiH, ShapiroIM and RisbudMV. (2015). Disc in flames: roles of TNF-α and IL-1β in intervertebral disc degeneration. Eur Cells Mater, 30:104–117.
21.
ShiC, DasV, LiX, KcR, QiuS, O-SullivanI, RipperRL, KroinJS, MwaleF, et al. (2018). Development of an in vivo mouse model of discogenic low back pain. J Cell Physiol, 233:6589–6602.
22.
VinceletteJ, XuY, ZhangLN, SchaeferCJ, VergonaR, SullivanME, HamptonTG and WangYX. (2007). Gait analysis in a murine model of collagen-induced arthritis. Arthritis Res Ther, 9:R123.
23.
MasudaK, AotaY, MuehlemanC, ImaiY, OkumaM, ThonarEJ, AnderssonGB and AnHS. (2005). A novel rabbit model of mild, reproducible disc degeneration by an anulus needle puncture: correlation between the degree of disc injury and radiological and histological appearances of disc degeneration. Spine (Phila Pa 1976), 30:5–14.
24.
WeiA, ShenB, WilliamsL and DiwanA. (2014). Mesenchymal stem cells: potential application in intervertebral disc regeneration. Transl Pediatr, 3:71.
25.
LiaoZ, LuoR, LiG, SongY, ZhanS, ZhaoK, HuaW, ZhangY, WuX and YangC. (2019). Exosomes from mesenchymal stem cells modulate endoplasmic reticulum stress to protect against nucleus pulposus cell death and ameliorate intervertebral disc degeneration in vivo. Theranostics, 9:4084–4100.
26.
WangT, LiP, MaX, TianP, HanC, ZangJ, KongJ and YanH. (2015). MicroRNA-494 inhibition protects nucleus pulposus cells from TNF-α-induced apoptosis by targeting JunD. Biochimie, 115:1–7.
27.
van den BoornJG, SchleeM, CochC and HartmannG. (2011). siRNA delivery with exosome nanoparticles. Nat Biotechnol, 29:325.
28.
ZhouY, ZhouG, TianC, JiangW, JinL, ZhangC and ChenX. (2016). Exosome-mediated small RNA delivery for gene therapy. Wiley Interdiscip Rev RNA, 7:758–771.
29.
van DommelenSM, VaderP, LakhalS, KooijmansS, van SolingeWW, WoodMJ and SchiffelersRM. (2012). Microvesicles and exosomes: opportunities for cell-derived membrane vesicles in drug delivery. J Control Release, 161:635–644.
30.
YeoRWY, LaiRC, ZhangB, TanSS, YinY, TehBJ and LimSK. (2013). Mesenchymal stem cell: an efficient mass producer of exosomes for drug delivery. Adv Drug Deliv Rev, 65:336–341.
ZhuJ, ZhangX, GaoW, HuH, WangX and HaoD. (2019). lncRNA/circRNA-miRNA-mRNA ceRNA network in lumbar intervertebral disc degeneration. Mol Med Rep, 20:3160–3174.
33.
ZhangY, YangJ, ZhouX, WangN, LiZ, ZhouY, FengJ, ShenD and ZhaoW. (2019). Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells. Int J Mol Med, 44:1357–1365.
34.
WangB, WangD, YanT and YuanH. (2016). MiR-138-5p promotes TNF-α-induced apoptosis in human intervertebral disc degeneration by targeting SIRT1 through PTEN/PI3K/Akt signaling. Exp Cell Res, 345:199–205.
35.
LadomeryM and DellaireG. (2002). Multifunctional zinc finger proteins in development and disease. Ann Hum Genet, 66:331–342.
36.
LuoA, ZhangX, FuL, ZhuZ and DongJ-T. (2016). Zinc finger factor ZNF121 is a MYC-interacting protein functionally affecting MYC and cell proliferation in epithelial cells. J Genet Genomics, 43:677–685.
37.
WuT, LinY and XieZ. (2018). MicroRNA-1247 inhibits cell proliferation by directly targeting ZNF346 in childhood neuroblastoma. Biol Res, 51:13.
38.
HuanC, XiaoxuC and XifangR. (2019). Zinc finger protein 521, negatively regulated by microRNA-204-5p, promotes proliferation, motility and invasion of gastric cancer cells. Technol Cancer Res Treat, 18:1533033819874783.
39.
AndaloussiSE, LakhalS, MägerI and WoodMJ. (2013). Exosomes for targeted siRNA delivery across biological barriers. Adv Drug Deliv Rev, 65:391–397.
40.
NaseriZ, OskueeRK, JaafariMR and MoghadamMF. (2018). Exosome-mediated delivery of functionally active miRNA-142-3p inhibitor reduces tumorigenicity of breast cancer in vitro and in vivo. Int J Nanomed, 13:7727.
41.
LuL, ChenX, TaoH, XiongW, JieS and LiH. (2015). Regulation of the expression of zinc finger protein genes by microRNAs enriched within acute lymphoblastic leukemia-derived microvesicles. Genet Mol Res, 14:11884–11895.
42.
NtziachristosV, BremerC and WeisslederR. (2003). Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging. Eur Radiol, 13:195–208.
43.
PetersonJD, LabrancheTP, VasquezKO, KossodoS, MeltonM, RaderR, ListelloJT, AbramsMA and MiskoTP. (2010). Optical tomographic imaging discriminates between disease-modifying anti-rheumatic drug (DMARD) and non-DMARD efficacy in collagen antibody-induced arthritis. Arthritis Res Ther, 12:R105.
RisbudMV and ShapiroIM. (2014). Role of cytokines in intervertebral disc degeneration: pain and disc content. Nat Rev Rheumatol, 10:44–56.
46.
ChengX, ZhangG, ZhangL, HuY, ZhangK, SunX, ZhaoC, LiH, LiYM and ZhaoJ. (2018). Mesenchymal stem cells deliver exogenous miR-21 via exosomes to inhibit nucleus pulposus cell apoptosis and reduce intervertebral disc degeneration. J Cell Mol Med, 22:261–276.
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