Both basal and submucosal gland (SMG) duct stem cells of the airway epithelium are capable of sphere formation in the in vitro sphere assay, although the efficiency at which this occurs is very low. We sought to improve this efficiency of sphere formation by identifying subpopulations of airway basal stem cells (ABSC) and SMG duct cells based on their aldehyde dehydrogenase (ALDH) activity. ALDHhi ABSCs and SMG duct cells were highly enriched for the population of cells that could make spheres, while the co-culture of ALDHhi differentiated cells with the ALDHhi ABSCs increased their sphere-forming efficiency. Specific ALDH agonists and antagonists were used to show that airway specific ALDH isozymes are important for ABSC proliferation. Pathway analysis of gene expression profiling of ALDHhi and ALDHlo ABSCs revealed a significant upregulation of the arachidonic acid (AA) metabolism pathway in ALDHhi ABSCs. We confirmed the importance of this pathway in the metabolism of proliferating ALDHhi ABSCs using bioenergetics studies as well as agonists and antagonists of the AA pathway. These studies could lead to the development of novel strategies for altering ABSC proliferation in the airway epithelium.
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References
1.
HegabAE, HaVL, GilbertJL, ZhangKX, MalkoskiSP, ChonAT, DarmawanDO, BishtB, OoiAT, et al. (2011). Novel stem/progenitor cell population from murine tracheal submucosal gland ducts with multipotent regenerative potential. Stem Cells, 29:1283–1293.
2.
HegabAE, HaVL, DarmawanDO, GilbertJL, OoiAT, AttigaYS, BishtB, NickersonDW and GompertsBN. (2012). Isolation and in vitro characterization of Basal and submucosal gland duct stem/progenitor cells from human proximal airways. Stem Cells Transl Med, 1:719–724.
3.
HegabAE, HaVL, AttigaYS, NickersonDW and GompertsBN. (2012). Isolation of basal cells and submucosal gland duct cells from mouse trachea. J Vis Exp. e3731.
4.
HongKU, ReynoldsSD, WatkinsS, FuchsE and StrippBR. (2004). Basal cells are a multipotent progenitor capable of renewing the bronchial epithelium. Am J Pathol, 164:577–588.
5.
RockJR, OnaitisMW, RawlinsEL, LuY, ClarkCP, XueY, RandellSH and HoganBL. (2009). Basal cells as stem cells of the mouse trachea and human airway epithelium. Proc Natl Acad Sci U S A, 106:12771–12775.
6.
PastranaE, Silva-VargasV and DoetschF. (2011). Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell Stem Cell, 8:486–498.
7.
ArmstrongL, StojkovicM, DimmickI, AhmadS, StojkovicP, HoleN and LakoM. (2004). Phenotypic characterization of murine primitive hematopoietic progenitor cells isolated on basis of aldehyde dehydrogenase activity. Stem Cells, 22:1142–1151.
8.
HessDA, WirthlinL, CraftTP, HerrbrichPE, HohmSA, LaheyR, EadesWC, CreerMH and NoltaJA. (2006). Selection based on CD133 and high aldehyde dehydrogenase activity isolates long-term reconstituting human hematopoietic stem cells. Blood, 107:2162–2169.
9.
GinestierC, HurMH, Charafe-JauffretE, MonvilleF, DutcherJ, BrownM, JacquemierJ, ViensP, KleerCG, et al. (2007). ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell, 1:555–567.
10.
PearceDJ, TaussigD, SimpsonC, AllenK, RohatinerAZ, ListerTA and BonnetD. (2005). Characterization of cells with a high aldehyde dehydrogenase activity from cord blood and acute myeloid leukemia samples. Stem Cells, 23:752–760.
11.
MarcatoP, DeanCA, GiacomantonioCA and LeePW. (2011). Aldehyde dehydrogenase: its role as a cancer stem cell marker comes down to the specific isoform. Cell Cycle, 10:1378–1384.
12.
PatelM, LuL, ZanderDS, SreeramaL, CocoD and MorebJS. (2008). ALDH1A1 and ALDH3A1 expression in lung cancers: correlation with histologic type and potential precursors. Lung Cancer, 59:340–349.
13.
SullivanJP, SpinolaM, DodgeM, RasoMG, BehrensC, GaoB, SchusterK, ShaoC, LarsenJE, et al. (2010). Aldehyde dehydrogenase activity selects for lung adenocarcinoma stem cells dependent on notch signaling. Cancer Res, 70:9937–9948.
14.
YoshidaA, RzhetskyA, HsuLC and ChangC. (1998). Human aldehyde dehydrogenase gene family. Eur J Biochem, 251:549–557.
15.
SophosNA and VasiliouV. (2003). Aldehyde dehydrogenase gene superfamily: the 2002 update. Chem Biol Interact, 143–144:5–22.
16.
VasiliouV and NebertDW. (2005). Analysis and update of the human aldehyde dehydrogenase (ALDH) gene family. Hum Genomics, 2:138–143.
17.
O'BrienPJ, SirakiAG and ShangariN. (2005). Aldehyde sources, metabolism, molecular toxicity mechanisms, and possible effects on human health. Crit Rev Toxicol, 35:609–662.
18.
YouY, RicherEJ, HuangT and BrodySL. (2002). Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population. Am J Physiol Lung Cell Mol Physiol, 283:L1315–L1321.
19.
TrapnellC, PachterL and SalzbergSL. (2009). TopHat: discovering splice junctions with RNA-Seq. Bioinformatics, 25:1105–1111.
20.
TrapnellC, WilliamsBA, PerteaG, MortazaviA, KwanG, van BarenMJ, SalzbergSL, WoldBJ and PachterL. (2010). Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol, 28:511–515.
21.
AndersS and HuberW. (2010). Differential expression analysis for sequence count data. Genome Biol, 11:R106
22.
FeigeJN, LagougeM, CantoC, StrehleA, HoutenSM, MilneJC, LambertPD, MatakiC, ElliottPJ and AuwerxJ. (2008). Specific SIRT1 activation mimics low energy levels and protects against diet-induced metabolic disorders by enhancing fat oxidation. Cell Metab, 8:347–358.
23.
CicaleseA, BonizziG, PasiCE, FarettaM, RonzoniS, GiuliniB, BriskenC, MinucciS, Di FiorePP and PelicciPG. (2009). The tumor suppressor p53 regulates polarity of self-renewing divisions in mammary stem cells. Cell, 138:1083–1095.
24.
AlnoutiY and KlaassenCD. (2008). Tissue distribution, ontogeny, and regulation of aldehyde dehydrogenase (Aldh) enzymes mRNA by prototypical microsomal enzyme inducers in mice. Toxicol Sci, 101:51–64.
25.
KeungWM and ValleeBL. (1993). Daidzin: a potent, selective inhibitor of human mitochondrial aldehyde dehydrogenase. Proc Natl Acad Sci U S A, 90:1247–1251.
26.
RenS, KalhornTF and SlatteryJT. (1999). Inhibition of human aldehyde dehydrogenase 1 by the 4-hydroxycyclophosphamide degradation product acrolein. Drug Metab Dispos, 27:133–137.
27.
CanutoRA, MaggioraM, TrombettaA, MartinassoG and MuzioG. (2003). Aldehyde dehydrogenase 3 expression is decreased by clofibrate via PPAR gamma induction in JM2 rat hepatoma cell line. Chem Biol Interact, 143–144:29–35.
28.
TsaoPN, VasconcelosM, IzvolskyKI, QianJ, LuJ and CardosoWV. (2009). Notch signaling controls the balance of ciliated and secretory cell fates in developing airways. Development, 136:2297–2307.
BanhA, XiaoN, CaoH, ChenCH, KuoP, KrakowT, BavanB, KhongB, YaoM, et al. (2011). A novel aldehyde dehydrogenase-3 activator leads to adult salivary stem cell enrichment in vivo. Clin Cancer Res, 17:7265–7272.
31.
KhannaM, ChenCH, Kimble-HillA, ParajuliB, Perez-MillerS, BaskaranS, KimJ, DriaK, VasiliouV, Mochly-RosenD and HurleyTD. (2011). Discovery of a novel class of covalent inhibitor for aldehyde dehydrogenases. J Biol Chem, 286:43486–43494.
32.
MorebJS, UcarD, HanS, AmoryJK, GoldsteinAS, OstmarkB and ChangLJ. (2012). The enzymatic activity of human aldehyde dehydrogenases 1A2 and 2 (ALDH1A2 and ALDH2) is detected by Aldefluor, inhibited by diethylaminobenzaldehyde and has significant effects on cell proliferation and drug resistance. Chem Biol Interact, 195:52–60.
33.
KatadaC, MutoM, NakayamaM, TanabeS, HiguchiK, SasakiT, AzumaM, IshidoK, KatadaN, et al. (2012). Risk of superficial squamous cell carcinoma developing in the head and neck region in patients with esophageal squamous cell carcinoma. Laryngoscope, 122:1291–1296.
34.
OyamaT, NagayoshiH, MatsudaT, OkaM, IsseT, YuHS, PhamTT, TanakaM, KagawaN, KanekoK and KawamotoT. (2010). Effects of acetaldehyde inhalation in mitochondrial aldehyde dehydrogenase deficient mice (Aldh2-/-). Front Biosci, 2:1344–1354.
35.
ParkJY, MatsuoK, SuzukiT, ItoH, HosonoS, KawaseT, WatanabeM, OzeI, HidaT, et al. (2010). Impact of smoking on lung cancer risk is stronger in those with the homozygous aldehyde dehydrogenase 2 null allele in a Japanese population. Carcinogenesis, 31:660–665.
36.
LewisSJ and SmithGD. (2005). Alcohol, ALDH2, and esophageal cancer: a meta-analysis which illustrates the potentials and limitations of a Mendelian randomization approach. Cancer Epidemiol Biomarkers Prev, 14:1967–1971.
37.
MatsuoK, OzeI, HosonoS, ItoH, WatanabeM, IshiokaK, ItoS, TajikaM, YatabeY, et al. (2013). The aldehyde dehydrogenase 2 (ALDH2) Glu504Lys polymorphism interacts with alcohol drinking in the risk of stomach cancer. Carcinogenesis, 34:1510–1515.
38.
BalberAE. (2011). Concise review: aldehyde dehydrogenase bright stem and progenitor cell populations from normal tissues: characteristics, activities, and emerging uses in regenerative medicine. Stem Cells, 29:570–575.
39.
SatoT, van EsJH, SnippertHJ, StangeDE, VriesRG, van den BornM, BarkerN, ShroyerNF, van de WeteringM and CleversH. (2011). Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature, 469:415–418.
40.
CummingsBS, McHowatJ and SchnellmannRG. (2000). Phospholipase A(2)s in cell injury and death. J Pharmacol Exp Ther, 294:793–799.
41.
SuraneniMV, Schneider-BroussardR, MooreJR, DavisTC, MaldonadoCJ, LiH, NewmanRA, KusewittD, HuJ, YangP and TangDG. (2010). Transgenic expression of 15-lipoxygenase 2 (15-LOX2) in mouse prostate leads to hyperplasia and cell senescence. Oncogene, 29:4261–4275.
42.
KenchegowdaS, BazanNG and BazanHE. (2011). EGF stimulates lipoxin A4 synthesis and modulates repair in corneal epithelial cells through ERK and p38 activation. Invest Ophthalmol Vis Sci, 52:2240–2249.
43.
AnderssonCK, ClaessonHE, Rydell-TormanenK, SwedmarkS, HallgrenA and ErjefaltJS. (2008). Mice lacking 12/15-lipoxygenase have attenuated airway allergic inflammation and remodeling. Am J Respir Cell Mol Biol, 39:648–656.
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