Evaluation of 309 Environmental Chemicals Using a Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity Assay

被引：47

作者：

Chandler, Kelly J. ^{[1
,2
]}

Barrier, Marianne ^{[1
]}

Jeffay, Susan ^{[1
]}

Nichols, Harriette P. ^{[1
]}

Kleinstreuer, Nicole C. ^{[2
]}

Singh, Amar V. ^{[3
]}

Reif, David M. ^{[2
]}

Sipes, Nisha S. ^{[2
]}

Judson, Richard S. ^{[2
]}

Dix, David J. ^{[2
]}

Kavlock, Robert ^{[2
]}

Hunter, Edward S., III ^{[1
]}

Knudsen, Thomas B. ^{[2
]}

机构：

[1] US EPA, Natl Hlth & Environm Effects Res Lab, Off Res & Dev, Res Triangle Pk, NC 27711 USA

[2] US EPA, Natl Ctr Computat Toxicol, Off Res & Dev, Res Triangle Pk, NC 27711 USA

[3] Lockheed Martin, Res Triangle Pk, NC USA

来源：

PLOS ONE | 2011年 / 6卷 / 06期

关键词：

VITRO EMBRYOTOXICITY TESTS; HYPOXIC PULMONARY-HYPERTENSION; ANTIOXIDANT RESPONSE ELEMENT; INDUCIBLE FACTOR 1-ALPHA; YA-SUBUNIT GENE; OXIDATIVE STRESS; TRANSCRIPTIONAL REGULATION; PHENOLIC ANTIOXIDANTS; PREDICTION MODELS; SELF-RENEWAL;

D O I：

10.1371/journal.pone.0018540

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The vast landscape of environmental chemicals has motivated the need for alternative methods to traditional whole-animal bioassays in toxicity testing. Embryonic stem (ES) cells provide an in vitro model of embryonic development and an alternative method for assessing developmental toxicity. Here, we evaluated 309 environmental chemicals, mostly food-use pesticides, from the ToxCast (TM) chemical library using a mouse ES cell platform. ES cells were cultured in the absence of pluripotency factors to promote spontaneous differentiation and in the presence of DMSO-solubilized chemicals at different concentrations to test the effects of exposure on differentiation and cytotoxicity. Cardiomyocyte differentiation (alpha, beta myosin heavy chain; MYH6/MYH7) and cytotoxicity (DRAQ5 (TM)/Sapphire700 (TM)) were measured by In-Cell Western (TM) analysis. Half-maximal activity concentration (AC(50)) values for differentiation and cytotoxicity endpoints were determined, with 18% of the chemical library showing significant activity on either endpoint. Mining these effects against the ToxCast Phase I assays (similar to 500) revealed significant associations for a subset of chemicals (26) that perturbed transcription-based activities and impaired ES cell differentiation. Increased transcriptional activity of several critical developmental genes including BMPR2, PAX6 and OCT1 were strongly associated with decreased ES cell differentiation. Multiple genes involved in reactive oxygen species signaling pathways (NRF2, ABCG2, GSTA2, HIF1A) were strongly associated with decreased ES cell differentiation as well. A multivariate model built from these data revealed alterations in ABCG2 transporter was a strong predictor of impaired ES cell differentiation. Taken together, these results provide an initial characterization of metabolic and regulatory pathways by which some environmental chemicals may act to disrupt ES cell growth and differentiation.

引用

页数：11

共 53 条

[1] BMP type II receptor is required for gastrulation and early development of mouse embryos [J].

Beppu, H ;

Kawabata, M ;

Hamamoto, T ;

Chytil, A ;

Minowa, O ;

Noda, T ;

Miyazono, K .

DEVELOPMENTAL BIOLOGY, 2000, 221 (01) :249-258

[2] Gene expression in human embryonic stem cell lines: unique molecular signature [J].

Bhattacharya, B ;

Miura, T ;

Brandenberger, R ;

Mejido, J ;

Luo, YQ ;

Yang, AX ;

Joshi, BH ;

Ginis, I ;

Thies, RS ;

Amit, M ;

Lyons, I ;

Condie, BG ;

Itskovitz-Eldor, J ;

Rao, MS ;

Puri, RK .

BLOOD, 2004, 103 (08) :2956-2964

[3] Maintenance of retinal stem cells by Abcg2 is regulated by notch signaling [J].

Bhattacharya, Sumitra ;

Das, Ani ;

Mallya, Kavita ;

Ahmad, Iqbal .

JOURNAL OF CELL SCIENCE, 2007, 120 (15) :2652-2662

[4] Loss of the Nrf2 transcription factor causes a marked reduction in constitutive and inducible expression of the glutathione S-transferase Gsta1, Gsta2, Gstm1, Gstm2, Gstm3 and Gstm4 genes in the livers of male and female mice [J].

Chanas, SA ;

Jiang, Q ;

McMahon, M ;

McWalter, GK ;

McLellan, LI ;

Elcombe, CR ;

Henderson, CJ ;

Wolf, CR ;

Moffat, GJ ;

Itoh, K ;

Yamamoto, M ;

Hayes, JD .

BIOCHEMICAL JOURNAL, 2002, 365 (02) :405-416

[5] Endless Possibilities: Stem Cells and the Vision for Toxicology Testing in the 21st Century [J].

Chapin, Robert E. ;

Stedman, Donald B. .

TOXICOLOGICAL SCIENCES, 2009, 112 (01) :17-22

[6] Toxicology - Transforming environmental health protection [J].

Collins, Francis S. ;

Gray, George M. ;

Bucher, John R. .

SCIENCE, 2008, 319 (5865) :906-907

[7] ABCG2: A potential marker of stem cells and novel target in stern cell and cancer therapy [J].

Ding, Xi-wei ;

Wu, Jun-hua ;

Jiang, Chun-ping .

LIFE SCIENCES, 2010, 86 (17-18) :631-637

[8]

FAVREAU LV, 1991, J BIOL CHEM, V266, P4556

[9] Increased susceptibility to hypoxic pulmonary hypertension in Bmpr2 mutant mice is associated with endothelial dysfunction in the pulmonary vasculature [J].

Frank, David B. ;

Lowery, Jonathan ;

Anderson, Lynda ;

Brink, Monique ;

Reese, Jeff ;

de Caestecker, Mark .

AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2008, 294 (01) :L98-L109

[10] XENOBIOTIC-INDUCIBLE EXPRESSION OF MURINE GLUTATHIONE-S-TRANSFERASE YA-SUBUNIT GENE IS CONTROLLED BY AN ELECTROPHILE-RESPONSIVE ELEMENT [J].

FRILING, RS ;

BENSIMON, A ;

TICHAUER, Y ;

DANIEL, V .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (16) :6258-6262

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