Bronchopulmonary dysplasia (BPD), the most common complication of extreme preterm birth, can be caused by oxygen-related lung injury and is characterized by impaired alveolar and vascular development. Mesenchymal stromal cells (MSCs) have lung protective effects. Conversely, BPD is associated with increased MSCs in tracheal aspirates. We hypothesized that endogenous lung (L-)MSCs are perturbed in a well-established oxygen-induced rat model mimicking BPD features. Rat pups were exposed to 21% or 95% oxygen from birth to postnatal day 10. On day 12, CD146+ L-MSCs were isolated and characterized according to the International Society for Cellular Therapy criteria. Epithelial and vascular repair potential were tested by scratch assay and endothelial network formation, respectively, immune function by mixed lymphocyte reaction assay. Microarray analysis was performed using the Affymetrix GeneChip and gene set enrichment analysis software. CD146+ L-MSCs isolated from rat pups exposed to hyperoxia had decreased CD73 expression and inhibited lung endothelial network formation. CD146+ L-MSCs indiscriminately promoted epithelial wound healing and limited T cell proliferation. Expression of potent antiangiogenic genes of the axonal guidance cue and CDC42 pathways was increased after in vivo hyperoxia, whereas genes of the anti-inflammatory Janus kinase (JAK)/signal transducer and activator of transcription (STAT) and lung/vascular growth-promoting fibroblast growth factor (FGF) pathways were decreased. In conclusion, in vivo hyperoxia exposure alters the proangiogenic effects and FGF expression of L-MSCs. In addition, decreased CD73 and JAK/STAT expression suggests decreased immune function. L-MSC function may be perturbed and contribute to BPD pathogenesis. These findings may lead to improvements in manufacturing exogenous MSCs with superior repair capabilities.
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References
1.
March of Dimes, PMNCH, Save the Children and WHO. (2012). Born Too Soon: The Global Action Report on Preterm Birth. HowsonC, KinneyM, LawnJ, eds. World Health Organization, Geneva.
2.
JobeAH and BancalariE. (2001). Bronchopulmonary dysplasia. Am J Respir Crit Care Med, 163:1723–1729.
3.
GreenoughA. (2012). Long term respiratory outcomes of very premature birth (<32 weeks). Semin Fetal Neonat Med, 17:73–76.
4.
FawkeJ, LumS, KirkbyJ, HennessyE, MarlowN, RowellV, ThomasS and StocksJ. (2010). Lung function and respiratory symptoms at 11 years in children born extremely preterm: the EPICure study. Am J Respir Crit Care Med, 182:237–245.
5.
WongPM, LeesAN, LouwJ, LeeFY, FrenchN, GainK, MurrayCP, WilsonA and ChambersDC. (2008). Emphysema in young adult survivors of moderate-to-severe bronchopulmonary dysplasia. Eur Respir J, 32:321–328.
6.
WHO. (2008). World Health Statistics 2008. TheakstonF, ed. World Health Organization, Geveva, Switzerland.
7.
SutskoRP, YoungKC, RibeiroA, TorresE, RodriguezM, HehreD, DeviaC, McNieceI and SuguiharaC. (2013). Long-term reparative effects of mesenchymal stem cell therapy following neonatal hyperoxia-induced lung injury. Pediatr Res, 73:46–53.
8.
ChangYS, OhW, ChoiSJ, SungDK, KimSY, ChoiEY, KangS, JinHJ, YangYS and ParkWS. (2009). Human umbilical cord blood-derived mesenchymal stem cells attenuate hyperoxia-induced lung injury in neonatal rats. Cell Transplant, 18:869–886.
9.
PierroM, IonescuL, MontemurroT, VadivelA, WeissmannG, OuditG, EmeryD, BodigaS, EatonF, et al. (2013). Short-term, long-term and paracrine effect of human umbilical cord-derived stem cells in lung injury prevention and repair in experimental bronchopulmonary dysplasia. Thorax, 68:475–484.
10.
van HaaftenT, ByrneR, BonnetS, RochefortGY, AkabutuJ, BouchentoufM, Rey-ParraGJ, GalipeauJ, HaromyA, et al. (2009). Airway delivery of mesenchymal stem cells prevents arrested alveolar growth in neonatal lung injury in rats. Am J Respir Crit Care Med, 180:1131–1142.
11.
AslamM, BavejaR, LiangOD, Fernandez-GonzalezA, LeeC, MitsialisSA and KourembanasS. (2009). Bone marrow stromal cells attenuate lung injury in a murine model of neonatal chronic lung disease. Am J Respir Crit Care Med, 180:1122–1130.
12.
LamaVN, SmithL, BadriL, FlintA, AndreiSM, WangZ, LiaoH, ToewsGB, KrebsbachPH, et al. (2007). Evidence for tissue-resident mesenchymal stem cells in human adult lung from studies of transplanted allografts. J Clin Invest, 117:989–996.
13.
HoffmanAM, PaxsonMR, MazanAMD, TyagiS, MurthyS, IngenitoEP, and IngenitoEP. (2011). Lung-derived mesenchymal stromal cell post-transplantation survival, persistence, paracrine expression, and repair of elastase-injured lung. Stem Cells Dev, 20:1779–1792.
14.
JarvinenL, BadriL, WettlauferS, OhtsukaT, StandifordTJ, ToewsGB, PinskyDJ, Peters-GoldenM and LamaVN. (2008). Lung resident mesenchymal stem cells isolated from human lung allografts inhibit T cell proliferation via a soluble mediator. J Immunol, 181:4389–4396.
15.
CollinsJJ and ThebaudB. (2014). Lung mesenchymal stromal cells in development and disease: to serve and protect?. Antioxid Redox Signal, 21:1849–1862.
16.
McQualterJL, McCartyRC, Van der VeldenJ, O'DonoghueRJ, Asselin-LabatML, BozinovskiS and BertoncelloI. (2013). TGF-beta signaling in stromal cells acts upstream of FGF-10 to regulate epithelial stem cell growth in the adult lung. Stem Cell Res, 11:1222–1233.
17.
ThebaudB, LadhaF, MichelakisED, SawickaM, ThurstonG, EatonF, HashimotoK, HarryG, HaromyA, KorbuttG and ArcherSL. (2005). Vascular endothelial growth factor gene therapy increases survival, promotes lung angiogenesis, and prevents alveolar damage in hyperoxia-induced lung injury: evidence that angiogenesis participates in alveolarization. Circulation, 112:2477–2486.
18.
CollinsJJ, MöbiusMA and ThébaudB. (2016). Isolation of CD146+ resident lung mesenchymal stromal cells from rat lungs. J Vis Exp 17:112.
19.
YaoQY, XuBL, WangJY, LiuHC, ZhangSC and TuCT. (2012). Inhibition by curcumin of multiple sites of the transforming growth factor-beta1 signalling pathway ameliorates the progression of liver fibrosis induced by carbon tetrachloride in rats. BMC Complement Altern Med, 12:156.
YamamotoY, BaldwinHS and PrinceLS. (2012). Endothelial differentiation by multipotent fetal mouse lung mesenchymal cells. Stem Cells Dev, 21:1455–1465.
22.
VadivelA, AlphonseRS, CollinsJJ, van HaaftenT, O'ReillyM, EatonF and ThebaudB. (2013). The axonal guidance cue semaphorin 3C contributes to alveolar growth and repair. PLoS One, 8:e67225.
23.
GebackT, SchulzMM, KoumoutsakosP and DetmarM. (2009). TScratch: a novel and simple software tool for automated analysis of monolayer wound healing assays. Biotechniques, 46:265–274.
24.
IonescuL, ByrneRN, van HaaftenT, VadivelA, AlphonseRS, Rey-ParraGJ, WeissmannG, HallA, EatonF and ThebaudB. (2012). Stem cell conditioned medium improves acute lung injury in mice: in vivo evidence for stem cell paracrine action. Am J Physiol Lung Cell Mol Physiol, 303:L967–L977.
25.
AlphonseRS, VadivelA, ColtanL, EatonF, BarrAJ, DyckJR and ThebaudB. (2011). Activation of Akt protects alveoli from neonatal oxygen-induced lung injury. Am J Respir Cell Mol Biol, 44:146–154.
26.
AlphonseRS, VadivelA, FungM, ShelleyWC, CritserPJ, IonescuL, O'ReillyM, OhlsRK, McConaghyS, et al. (2014). Existence, functional impairment, and lung repair potential of endothelial colony-forming cells in oxygen-induced arrested alveolar growth. Circulation, 129:2144–2157.
27.
SeghezziG, PatelS, RenCJ, GualandrisA, PintucciG, RobbinsES, ShapiroRL, GallowayAC, RifkinDB and MignattiP. (1998). Fibroblast growth factor-2 (FGF-2) induces vascular endothelial growth factor (VEGF) expression in the endothelial cells of forming capillaries: an autocrine mechanism contributing to angiogenesis. J Cell Biol, 141:1659–1673.
28.
MuulLM, HeineG, SilvinC, JamesSP, CandottiF, RadbruchA and WormM. (2011). Measurement of proliferative responses of cultured lymphocytes. Curr Protoc Immunol Chapter 7: Unit 7.10.
29.
WeissML, AndersonC, MedicettyS, SeshareddyKB, WeissRJ, VanderWerffI, TroyerD and McIntoshKR. (2008). Immune properties of human umbilical cord ’Wharton's jelly-derived cells. Stem Cells, 26:2865–2874.
30.
KellyE, WonA, RefaeliY and Van ParijsL. (2002). IL-2 and related cytokines can promote T cell survival by activating AKT. J Immunol, 168:597–603.
31.
RoedererM. (2011). Interpretation of cellular proliferation data: avoid the Panglossian. Cytometry A, 79:95–101.
32.
SubramanianA, TamayoP, MoothaVK, MukherjeeS, EbertBL, GilletteMA, PaulovichA, PomeroySL, GolubTR, LanderES and MesirovJP. (2005). Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A, 102:15545–15550.
33.
MoothaVK, LindgrenCM, ErikssonKF, SubramanianA, SihagS, LeharJ, PuigserverP, CarlssonE, RidderstraleM, et al. (2003). PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet, 34:267–273.
34.
dos SantosCC, MurthyS, HuP, ShanY, HaitsmaJJ, MeiSH, StewartDJ and LilesWC. (2012). Network analysis of transcriptional responses induced by mesenchymal stem cell treatment of experimental sepsis. Am J Pathol, 181:1681–1692.
35.
ShannonP, MarkielA, OzierO, BaligaNS, WangJT, RamageD, AminN, SchwikowskiB and IdekerT. (2003). Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res, 13:2498–2504.
36.
BatalhaVL, FerreiraDG, CoelhoJE, ValadasJS, GomesR, Temido-FerreiraM, ShmidtT, BaqiY, BueeL, et al. (2016). The caffeine-binding adenosine A2A receptor induces age-like HPA-axis dysfunction by targeting glucocorticoid receptor function. Sci Rep, 6:31493.
37.
HuH, XuanY, XueM, ChengW, WangY, LiX, YinJ, LiX, YangN, ShiY and YanS. (2016). Semaphorin 3A attenuates cardiac autonomic disorders and reduces inducible ventricular arrhythmias in rats with experimental myocardial infarction. BMC Cardiovasc Disord, 16:16.
38.
ToyodaK, OkanoH, BambaH, HisaY, OomuraY, ImamuraT, FurukawaS and TooyamaI. (2006). Comparison of FGF1 (aFGF) expression between the dorsal motor nucleus of vagus and the hypoglossal nucleus of rat. Acta Histochem Cytochem, 39:1–7.
39.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
40.
KramperaM, GalipeauJ, ShiY, TarteK, SensebeL and MSC. (2013). Immunological characterization of multipotent mesenchymal stromal cells–The International Society for Cellular Therapy (ISCT) working proposal. Cytotherapy, 15:1054–1061.
41.
ZhangCL, ZouY, HeW, GageFH and EvansRM. (2008). A role for adult TLX-positive neural stem cells in learning and behaviour. Nature, 451:1004–1007.
42.
MullerFJ, LaurentLC, KostkaD, UlitskyI, WilliamsR, LuC, ParkIH, RaoMS, ShamirR, et al. (2008). Regulatory networks define phenotypic classes of human stem cell lines. Nature, 455:401–405.
43.
WongDJ, LiuH, RidkyTW, CassarinoD, SegalE and ChangHY. (2008). Module map of stem cell genes guides creation of epithelial cancer stem cells. Cell Stem Cell, 2:333–344.
44.
LabbeE, LockL, LetamendiaA, GorskaAE, GryfeR, GallingerS, MosesHL and AttisanoL. (2007). Transcriptional cooperation between the transforming growth factor-beta and Wnt pathways in mammary and intestinal tumorigenesis. Cancer Res, 67:75–84.
45.
De BosscherK, Vanden BergheW and HaegemanG. (2006). Cross-talk between nuclear receptors and nuclear factor kappaB. Oncogene, 25:6868–6886.
46.
WestonGC, HavivI and RogersPA. (2002). Microarray analysis of VEGF-responsive genes in myometrial endothelial cells. Mol Hum Reprod, 8:855–863.
47.
BurtonGR, GuanY, NagarajanR and McGeheeREJr. (2002). Microarray analysis of gene expression during early adipocyte differentiation. Gene, 293:21–31.
48.
BoquestAC, ShahdadfarA, FronsdalK, SigurjonssonO, TunheimSH, CollasP and BrinchmannJE. (2005). Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture. Mol Biol Cell, 16:1131–1141.
49.
AltemeierWA, Matute-BelloG, GharibSA, GlennyRW, MartinTR and LilesWC. (2005). Modulation of lipopolysaccharide-induced gene transcription and promotion of lung injury by mechanical ventilation. J Immunol, 175:3369–3376.
50.
AltE, YanY, GehmertS, SongYH, AltmanA, GehmertS, VykoukalD and BaiX. (2011). Fibroblasts share mesenchymal phenotypes with stem cells, but lack their differentiation and colony-forming potential. Biol Cell, 103:197–208.
51.
Rolandsson EnesS, Andersson SjolandA, SkogI, HanssonL, LarssonH, Le BlancK, ErikssonL, BjermerL, SchedingS and Westergren-ThorssonG. (2016). MSC from fetal and adult lungs possess lung-specific properties compared to bone marrow-derived MSC. Sci Rep, 6:29160.
52.
GaskillCF, CarrierEJ, KropskiNC, BloodworthSM, ForonjyRF, TaketoMM, HongCC, AustinED, WestJD, et al. (2017). Disruption of lineage specification in adult pulmonary mesenchymal progenitor cells promotes microvascular dysfunction. J Clin Invest, 127:2262–2276.
53.
GaskillC, MarriottS, PratapS, MenonS, HedgesLK, FesselJP, KropskiD, AmesLW, LoydJE, et al. (2016). Shared gene expression patterns in mesenchymal progenitors derived from lung and epidermis in pulmonary arterial hypertension: identifying key pathways in pulmonary vascular disease. Pulm Circ, 6:483–497.
54.
KagoshimaM and ItoT. (2001). Diverse gene expression and function of semaphorins in developing lung: positive and negative regulatory roles of semaphorins in lung branching morphogenesis. Genes Cells, 6:559–571.
55.
VadivelA, van HaaftenT, AlphonseRS, Rey-ParraGJ, IonescuL, HaromyA, EatonF, MichelakisE and ThebaudB. (2012). Critical role of the axonal guidance cue EphrinB2 in lung growth, angiogenesis, and repair. Am J Respir Crit Care Med, 185:564–574.
56.
MaioneF, MollaF, MedaC, LatiniR, ZentilinL, GiaccaM, SeanoG, SeriniG, BussolinoF and GiraudoE. (2009). Semaphorin 3A is an endogenous angiogenesis inhibitor that blocks tumor growth and normalizes tumor vasculature in transgenic mouse models. J Clin Invest, 119:3356–3372.
57.
CasazzaA, KigelB, MaioneF, CapparucciaL, KesslerO, GiraudoE, MazzoneM, NeufeldG and TamagnoneL. (2012). Tumour growth inhibition and anti-metastatic activity of a mutated furin-resistant Semaphorin 3E isoform. EMBO Mol Med, 4:234–250.
58.
CasazzaA, FuX, JohanssonI, CapparucciaL, AnderssonF, GiustacchiniA, SquadritoML, VenneriMA, MazzoneM, et al. (2011). Systemic and targeted delivery of semaphorin 3A inhibits tumor angiogenesis and progression in mouse tumor models. Arterioscler Thromb Vasc Biol, 31:741–749.
59.
NeufeldG and KesslerO. (2008). The semaphorins: versatile regulators of tumour progression and tumour angiogenesis. Nat Rev Cancer, 8:632–645.
60.
WhiteAC, LavineKJ and OrnitzDM. (2007). FGF9 and SHH regulate mesenchymal Vegfa expression and development of the pulmonary capillary network. Development, 134:3743–3752.
61.
KwonHM, HurSM, ParkKY, KimCK, KimYM, KimHS, ShinHC, WonMH, HaKS, et al. (2014). Multiple paracrine factors secreted by mesenchymal stem cells contribute to angiogenesis. Vascul Pharmacol, 63:19–28.
62.
AlviraCM. (2016). Aberrant pulmonary vascular growth and remodeling in bronchopulmonary dysplasia. Front Med (Lausanne), 3:21.
63.
GuX, KarpPH, BrodySL, PierceRA, WelshMJ, HoltzmanMJ and Ben-ShaharY. (2014). Chemosensory functions for pulmonary neuroendocrine cells. Am J Respir Cell Mol Biol, 50:637–646.
64.
PluznickJL, ProtzkoRJ, GevorgyanH, PeterlinZ, SiposA, HanJ, BrunetI, WanLX, ReyF, et al. (2013). Olfactory receptor responding to gut microbiota-derived signals plays a role in renin secretion and blood pressure regulation. Proc Natl Acad Sci U S A, 110:4410–4415.
65.
RothJM, KohlerD, SchneiderM, GranjaTF and RosenbergerP. (2016). Semaphorin 7A Aggravates Pulmonary Inflammation during Lung Injury. PLoS One, 11:e0146930.
66.
WhiteAC, XuJ, YinY, SmithC, SchmidG and OrnitzDM. (2006). FGF9 and SHH signaling coordinate lung growth and development through regulation of distinct mesenchymal domains. Development, 133:1507–1517.
67.
BostromH, WillettsK, PeknyM, LeveenP, LindahlP, HedstrandH, PeknaM, HellstromM, Gebre-MedhinS, et al. (1996). PDGF-A signaling is a critical event in lung alveolar myofibroblast development and alveogenesis. Cell, 85:863–873.
68.
Al AlamD, El AghaE, SakuraiR, KheirollahiV, MoiseenkoA, DanopoulosS, ShresthaA, SchmoldtC, QuantiusJ, et al. (2015). Evidence for the involvement of fibroblast growth factor 10 in lipofibroblast formation during embryonic lung development. Development, 142:4139–4150.
69.
LindahlP, KarlssonL, HellstromM, Gebre-MedhinS, WillettsK, HeathJK and BetsholtzC. (1997). Alveogenesis failure in PDGF-A-deficient mice is coupled to lack of distal spreading of alveolar smooth muscle cell progenitors during lung development. Development, 124:3943–3953.
70.
PerlAK and GaleE. (2009). FGF signaling is required for myofibroblast differentiation during alveolar regeneration. Am J Physiol Lung Cell Mol Physiol, 297:L299–L308.
71.
El AghaE, HeroldS, AlamDAl, QuantiusJ, MacKenzieB, CarraroG, MoiseenkoA, ChaoCM, MinooP, SeegerW and BellusciS. (2014). Fgf10-positive cells represent a progenitor cell population during lung development and postnatally. Development, 141:296–306.
72.
ChaoCM, MoiseenkoA, ZimmerKP and BellusciS. (2016). Alveologenesis: key cellular players and fibroblast growth factor 10 signaling. Mol Cell Pediatr, 3:17.
73.
ChaoCM, YahyaF, MoiseenkoA, TiozzoC, ShresthaA, AhmadvandN, El AghaE, QuantiusJ, DilaiS, et al. (2017). Fgf10 deficiency is causative for lethality in a mouse model of bronchopulmonary dysplasia. J Pathol, 241:91–103.
74.
BenjaminJT, CarverBJ, PlosaEJ, YamamotoY, MillerJD, LiuJH, van der MeerR, BlackwellTS and PrinceLS. (2010). NF-kappaB activation limits airway branching through inhibition of Sp1-mediated fibroblast growth factor-10 expression. J Immunol, 185:4896–4903.
75.
BenjaminJT, SmithRJ, HalloranBA, DayTJ, KellyDR and PrinceLS. (2007). FGF-10 is decreased in bronchopulmonary dysplasia and suppressed by Toll-like receptor activation. Am J Physiol Lung Cell Mol Physiol, 292:L550–L558.
76.
AcostaJM, ThebaudB, CastilloC, MailleuxA, TefftD, WuenschellC, AndersonKD, BourbonJ, ThieryJP, BellusciS and WarburtonD. (2001). Novel mechanisms in murine nitrofen-induced pulmonary hypoplasia: FGF-10 rescue in culture. Am J Physiol Lung Cell Mol Physiol, 281:L250–L257.
77.
PopovaAP, BentleyJK, CuiTX, RichardsonMN, LinnMJ, LeiJ, ChenQ, GoldsmithAM, PryhuberGS and HershensonMB. (2014). Reduced platelet-derived growth factor receptor expression is a primary feature of human bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol, 307:L231–L239.
78.
CaplanAI and SorrellJM. (2015). The MSC curtain that stops the immune system. Immunol Lett, 168:136–139.
79.
CaplanAI and CorreaD. (2011). The MSC: an injury drugstore. Cell Stem Cell, 9:11–15.
80.
Nogueira-SilvaC, PiairoP, Carvalho-DiasE, VeigaC, MouraRS and Correia-PintoJ. (2013). The role of glycoprotein 130 family of cytokines in fetal rat lung development. PLoS One, 8:e67607.
81.
BelizarioJE, Fontes-OliveiraCC, BorgesJP, KashiabaraJA and VannierE.. (2016). Skeletal muscle wasting and renewal: a pivotal role of myokine IL-6. Springerplus. 5:619.
82.
LinG, ZhangH, SunF, LuZ, Reed-MaldonadoA, LeeYC, WangG, BanieL and LueTF. (2016). Brain-derived neurotrophic factor promotes nerve regeneration by activating the JAK/STAT pathway in Schwann cells. Transl Androl Urol, 5:167–175.
83.
RicardN, TuL, Le HiressM, HuertasA, PhanC, ThuilletR, SattlerC, FadelE, SeferianA, et al. (2014). Increased pericyte coverage mediated by endothelial-derived fibroblast growth factor-2 and interleukin-6 is a source of smooth muscle-like cells in pulmonary hypertension. Circulation, 129:1586–1597.
84.
VarinA and GordonS. (2009). Alternative activation of macrophages: immune function and cellular biology. Immunobiology, 214:630–641.
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