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Article: Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development.

Smith, Brenden W / Stanford, Elizabeth A / Sherr, David H / Murphy, George J

2016 Volume 2016, Page(s) 2574152

Abstract: The aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that increases the expression of detoxifying enzymes upon ligand stimulation. Recent studies now suggest that novel endogenous roles of the AHR exist throughout development. ... ...

Abstract	The aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that increases the expression of detoxifying enzymes upon ligand stimulation. Recent studies now suggest that novel endogenous roles of the AHR exist throughout development. In an effort to create an optimized model system for the study of AHR signaling in several cellular lineages, we have employed a CRISPR/CAS9 genome editing strategy in induced pluripotent stem cells (iPSCs) to incorporate a reporter cassette at the transcription start site of one of its canonical targets, cytochrome P450 1A1 (CYP1A1). This cell line faithfully reports on CYP1A1 expression, with luciferase levels as its functional readout, when treated with an endogenous AHR ligand (FICZ) at escalating doses. iPSC-derived fibroblast-like cells respond to acute exposure to environmental and endogenous AHR ligands, and iPSC-derived hepatocytes increase CYP1A1 in a similar manner to primary hepatocytes. This cell line is an important innovation that can be used to map AHR activity in discrete cellular subsets throughout developmental ontogeny. As further endogenous ligands are proposed, this line can be used to screen for safety and efficacy and can report on the ability of small molecules to regulate critical cellular processes by modulating the activity of the AHR.
Language	English
Publishing date	2016-04-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	2573856-2
ISSN	1687-9678 ; 1687-966X
ISSN (online)	1687-9678
ISSN	1687-966X
DOI	10.1155/2016/2574152
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Article ; Online: Dioxins and related environmental contaminants increase TDP-43 levels.

Ash, Peter E A / Stanford, Elizabeth A / Al Abdulatif, Ali / Ramirez-Cardenas, Alejandra / Ballance, Heather I / Boudeau, Samantha / Jeh, Amanda / Murithi, James M / Tripodis, Yorghos / Murphy, George J / Sherr, David H / Wolozin, Benjamin

Molecular neurodegeneration

2017 Volume 12, Issue 1, Page(s) 35

Abstract: Background: Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative condition that is characterized by progressive loss of motor neurons and the accumulation of aggregated TAR DNA Binding Protein-43 (TDP-43, gene: TARDBP). Increasing ... ...

Abstract	Background: Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative condition that is characterized by progressive loss of motor neurons and the accumulation of aggregated TAR DNA Binding Protein-43 (TDP-43, gene: TARDBP). Increasing evidence indicates that environmental factors contribute to the risk of ALS. Dioxins, related planar polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants that activate the aryl hydrocarbon receptor (AHR), a ligand-activated, PAS family transcription factor. Recently, exposure to these toxicants was identified as a risk factor for ALS. Methods: We examined levels of TDP-43 reporter activity, transcript and protein. Quantification was done using cell lines, induced pluripotent stem cells (iPSCs) and mouse brain. The target samples were treated with AHR agonists, including 6-Formylindolo[3,2-b]carbazole (FICZ, a potential endogenous ligand, 2,3,7,8-tetrachlorodibenzo(p)dioxin, and benzo(a)pyrene, an abundant carcinogen in cigarette smoke). The action of the agonists was inhibited by concomitant addition of AHR antagonists or by AHR-specific shRNA. Results: We now report that AHR agonists induce up to a 3-fold increase in TDP-43 protein in human neuronal cell lines (BE-M17 cells), motor neuron differentiated iPSCs, and in murine brain. Chronic treatment with AHR agonists elicits over 2-fold accumulation of soluble and insoluble TDP-43, primarily because of reduced TDP-43 catabolism. AHR antagonists or AHR knockdown inhibits agonist-induced increases in TDP-43 protein and TARDBP transcription demonstrating that the ligands act through the AHR. Conclusions: These results provide the first evidence that environmental AHR ligands increase TDP-43, which is the principle pathological protein associated with ALS. These results suggest novel molecular mechanisms through which a variety of prevalent environmental factors might directly contribute to ALS. The widespread distribution of dioxins, PCBs and PAHs is considered to be a risk factor for cancer and autoimmune diseases, but could also be a significant public health concern for ALS.
MeSH term(s)	Amyotrophic Lateral Sclerosis ; Animals ; Brain/drug effects ; Cell Line ; DNA-Binding Proteins/biosynthesis ; DNA-Binding Proteins/drug effects ; Environmental Pollutants/adverse effects ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Neurons/drug effects ; Polychlorinated Dibenzodioxins/adverse effects ; Polycyclic Aromatic Hydrocarbons/adverse effects ; Receptors, Aryl Hydrocarbon/agonists
Chemical Substances	DNA-Binding Proteins ; Environmental Pollutants ; Polychlorinated Dibenzodioxins ; Polycyclic Aromatic Hydrocarbons ; Receptors, Aryl Hydrocarbon ; TARDBP protein, human ; TDP-43 protein, mouse
Language	English
Publishing date	2017-05-05
Publishing country	England
Document type	Journal Article
ZDB-ID	2244557-2
ISSN	1750-1326 ; 1750-1326
ISSN (online)	1750-1326
ISSN	1750-1326
DOI	10.1186/s13024-017-0177-9
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Article ; Online: The frequency, trajectories and predictors of adolescent recurrent pain: a population-based approach.

Stanford, Elizabeth A / Chambers, Christine T / Biesanz, Jeremy C / Chen, Edith

Pain

2008 Volume 138, Issue 1, Page(s) 11–21

Abstract: Recurrent pains are a complex set of conditions that cause great discomfort and impairment in children and adults. The objectives of this study were to (a) describe the frequency of headache, stomachache, and backache in a representative Canadian ... ...

Abstract	Recurrent pains are a complex set of conditions that cause great discomfort and impairment in children and adults. The objectives of this study were to (a) describe the frequency of headache, stomachache, and backache in a representative Canadian adolescent sample and (b) determine whether a set of psychosocial factors, including background factors (i.e., sex, pubertal status, parent chronic pain), external events (i.e., injury, illness/hospitalization, stressful-life events), and emotional factors (i.e., anxiety/depression, self-esteem) were predictive of these types of recurrent pain. Statistics Canada's National Longitudinal Survey of Children and Youth was used to assess a cohort of 2488 10- to 11-year-old adolescents up to five times, every 2 years. Results showed that, across 12-19 years of age, weekly or more frequent rates ranged from 26.1%-31.8% for headache, 13.5-22.2% for stomachache, and 17.6-25.8% for backache. Chi-square tests indicated that girls had higher rates of pain than boys for all types of pain, at all time points. Structural equation modeling using latent growth curves showed that sex and anxiety/depression at age 10-11 years was predictive of the start- and end-point intercepts (i.e., trajectories that indicated high levels of pain across time) and/or slopes (i.e., trajectories of pain that increased over time) for all three types of pain. Although there were also other factors that predicted only certain pain types or certain trajectory types, overall the results of this study suggest that adolescent recurrent pain is very common and that psychosocial factors can predict trajectories of recurrent pain over time across adolescence.
MeSH term(s)	Adolescent ; Adult ; Age Distribution ; Canada/epidemiology ; Child ; Female ; Humans ; Longitudinal Studies ; Male ; Pain/diagnosis ; Pain/epidemiology ; Population Dynamics ; Prevalence ; Risk Assessment/methods ; Risk Factors ; Sex Distribution
Language	English
Publishing date	2008-02-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	193153-2
ISSN	1872-6623 ; 0304-3959
ISSN (online)	1872-6623
ISSN	0304-3959
DOI	10.1016/j.pain.2007.10.032
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Article: A normative analysis of the development of pain-related vocabulary in children.

Stanford, Elizabeth A / Chambers, Christine T / Craig, Kenneth D

Pain

2005 Volume 114, Issue 1-2, Page(s) 278–284

Abstract: Effective verbalization of pain requires progressive cognitive development and acquisition of social communication skills. Use of self-report in pediatric pain assessment assumes children have acquired a capacity to understand and use common words to ... ...

Abstract	Effective verbalization of pain requires progressive cognitive development and acquisition of social communication skills. Use of self-report in pediatric pain assessment assumes children have acquired a capacity to understand and use common words to describe pain. The current investigation documented the language most commonly used by young children to describe pain and the age of onset of use of these words. Two complementary research methodologies were employed. Study 1 used the CHILDES database, an aggregated transcript database of multiple research studies examining spontaneous speech development across childhood. Transcripts of 14 randomly selected studies, yielding a total of 245 child participants ranging in age from 1 to 9 years, were searched for seven English primary pain word-stems: 'ache', 'boo-boo', 'hurt', 'ouch', 'ow', 'pain', and 'sore'. Study 2 surveyed 111 parents of children aged 3 to 6 years old concerning words the children commonly used for pain. Parents rated their children's frequency and age of first use of the seven pain word-stems. Both studies indicated that the most frequently used word-stems were 'hurt', 'ouch', and 'ow'. These words first emerged in children's vocabularies as early as 18 months of age. The word-stem 'pain' was used relatively infrequently and gradually emerged in children's vocabularies. The findings indicate that young children rely on a select number of words to describe pain, with these words appearing in children's vocabularies at an early age. These results have implications for developmentally appropriate pain assessment in young children.
MeSH term(s)	Child ; Child, Preschool ; Databases, Factual ; Female ; Humans ; Infant ; Male ; Pain ; Surveys and Questionnaires ; Vocabulary
Language	English
Publishing date	2005-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	193153-2
ISSN	1872-6623 ; 0304-3959
ISSN (online)	1872-6623
ISSN	0304-3959
DOI	10.1016/j.pain.2004.12.029
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Article: The role of developmental factors in predicting young children's use of a self-report scale for pain.

Stanford, Elizabeth A / Chambers, Christine T / Craig, Kenneth D

Pain

2005 Volume 120, Issue 1-2, Page(s) 16–23

Abstract: Accurate pain assessment is the foundation for effective pain management in children. At present, there is no clear consensus regarding the age at which young children are able to appropriately use self-report scales for pain. This study examined young ... ...

Abstract	Accurate pain assessment is the foundation for effective pain management in children. At present, there is no clear consensus regarding the age at which young children are able to appropriately use self-report scales for pain. This study examined young children's ability to use the Faces Pain Scale-Revised; (FPS-R; [Hicks CL, von Baeyer CL, Spafford PA, van Korlaar I, Goodenough B. The Faces Pain Scale-Revised: toward a common metric in pediatric pain measurement. Pain 2001; 93: 173-83]) for pain in response to vignettes and investigated the role of developmental factors in predicting their ability to use the scale. One hundred and twelve healthy children (3-6 years old) were assessed for their ability to accurately use a common faces scale to rate pain in hypothetical vignettes depicting pain scenarios common in childhood. Accuracy was determined by considering whether children's judgements of pain severity matched the pain severity depicted in the various vignettes. Children were also administered measures of numerical reasoning, language, and overall cognitive development. Results indicated that 5- and 6-year-old children were significantly more accurate in their use of the FPS-R in response to the vignettes than 4-year-old children, who in turn were significantly more accurate than 3-year-old children. However, over half of the 6-year-olds demonstrated difficulties using the FPS-R in response to the vignettes. Child age was the only significant predictor of children's ability to use the scale in response to the vignettes. Thus, a substantial number of young children experienced difficulties using the FPS-R when rating pain in hypothetical vignettes, although the ability to use the scale did improve with age.
MeSH term(s)	Age Distribution ; Canada/epidemiology ; Child ; Child, Preschool ; Facial Expression ; Female ; Humans ; Male ; Pain/classification ; Pain/diagnosis ; Pain Measurement/methods ; Pain Measurement/statistics & numerical data ; Reproducibility of Results ; Sensitivity and Specificity ; Severity of Illness Index ; Surveys and Questionnaires
Language	English
Publishing date	2005-12-13
Publishing country	United States
Document type	Journal Article ; Randomized Controlled Trial ; Research Support, Non-U.S. Gov't
ZDB-ID	193153-2
ISSN	1872-6623 ; 0304-3959
ISSN (online)	1872-6623
ISSN	0304-3959
DOI	10.1016/j.pain.2005.10.004
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Article ; Online: An Aryl Hydrocarbon Receptor-Mediated Amplification Loop That Enforces Cell Migration in ER-/PR-/Her2- Human Breast Cancer Cells.

Novikov, Olga / Wang, Zhongyan / Stanford, Elizabeth A / Parks, Ashley J / Ramirez-Cardenas, Alejandra / Landesman, Esther / Laklouk, Israa / Sarita-Reyes, Carmen / Gusenleitner, Daniel / Li, Amy / Monti, Stefano / Manteiga, Sara / Lee, Kyongbum / Sherr, David H

Molecular pharmacology

2016 Volume 90, Issue 5, Page(s) 674–688

Abstract: The endogenous ligand-activated aryl hydrocarbon receptor (AHR) plays an important role in numerous biologic processes. As the known number of AHR-mediated processes grows, so too does the importance of determining what endogenous AHR ligands are ... ...

Abstract	The endogenous ligand-activated aryl hydrocarbon receptor (AHR) plays an important role in numerous biologic processes. As the known number of AHR-mediated processes grows, so too does the importance of determining what endogenous AHR ligands are produced, how their production is regulated, and what biologic consequences ensue. Consequently, our studies were designed primarily to determine whether ER
MeSH term(s)	Cell Line, Tumor ; Cell Movement ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; Immunohistochemistry ; Kynurenine/metabolism ; Ligands ; Models, Biological ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Receptor, ErbB-2/metabolism ; Receptors, Aryl Hydrocarbon/genetics ; Receptors, Aryl Hydrocarbon/metabolism ; Receptors, Estrogen/metabolism ; Receptors, Progesterone/metabolism ; Triple Negative Breast Neoplasms/genetics ; Triple Negative Breast Neoplasms/metabolism ; Triple Negative Breast Neoplasms/pathology ; Tryptophan/metabolism ; Tryptophan Oxygenase/genetics ; Tryptophan Oxygenase/metabolism ; Xanthurenates/metabolism
Chemical Substances	Ligands ; RNA, Messenger ; Receptors, Aryl Hydrocarbon ; Receptors, Estrogen ; Receptors, Progesterone ; Xanthurenates ; Kynurenine (343-65-7) ; xanthurenic acid (58LAB1BG8J) ; Tryptophan (8DUH1N11BX) ; Tryptophan Oxygenase (EC 1.13.11.11) ; ERBB2 protein, human (EC 2.7.10.1) ; Receptor, ErbB-2 (EC 2.7.10.1)
Language	English
Publishing date	2016-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	124034-1
ISSN	1521-0111 ; 0026-895X
ISSN (online)	1521-0111
ISSN	0026-895X
DOI	10.1124/mol.116.105361
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Zs.A 525: Show issues

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Article ; Online: The role of the aryl hydrocarbon receptor in the development of cells with the molecular and functional characteristics of cancer stem-like cells.

Stanford, Elizabeth A / Wang, Zhongyan / Novikov, Olga / Mulas, Francesca / Landesman-Bollag, Esther / Monti, Stefano / Smith, Brenden W / Seldin, David C / Murphy, George J / Sherr, David H

BMC biology

2016 Volume 14, Page(s) 20

Abstract: Background: Self-renewing, chemoresistant breast cancer stem cells are believed to contribute significantly to cancer invasion, migration and patient relapse. Therefore, the identification of signaling pathways that regulate the acquisition of stem-like ...

Abstract	Background: Self-renewing, chemoresistant breast cancer stem cells are believed to contribute significantly to cancer invasion, migration and patient relapse. Therefore, the identification of signaling pathways that regulate the acquisition of stem-like qualities is an important step towards understanding why patients relapse and towards development of novel therapeutics that specifically target cancer stem cell vulnerabilities. Recent studies identified a role for the aryl hydrocarbon receptor (AHR), an environmental carcinogen receptor implicated in cancer initiation, in normal tissue-specific stem cell self-renewal. These studies inspired the hypothesis that the AHR plays a role in the acquisition of cancer stem cell-like qualities. Results: To test this hypothesis, AHR activity in Hs578T triple negative and SUM149 inflammatory breast cancer cells were modulated with AHR ligands, shRNA or AHR-specific inhibitors, and phenotypic, genomic and functional stem cell-associated characteristics were evaluated. The data demonstrate that (1) ALDH(high) cells express elevated levels of Ahr and Cyp1b1 and Cyp1a1, AHR-driven genes, (2) AHR knockdown reduces ALDH activity by 80%, (3) AHR hyper-activation with several ligands, including environmental ligands, significantly increases ALDH1 activity, expression of stem cell- and invasion/migration-associated genes, and accelerates cell migration, (4) a significant correlation between Ahr or Cyp1b1 expression (as a surrogate marker for AHR activity) and expression of stem cell- and invasion/migration-associated gene sets is seen with genomic data obtained from 79 human breast cancer cell lines and over 1,850 primary human breast cancers, (5) the AHR interacts directly with Sox2, a master regulator of self-renewal; AHR ligands increase this interaction and nuclear SOX2 translocation, (6) AHR knockdown inhibits tumorsphere formation in low adherence conditions, (7) AHR inhibition blocks the rapid migration of ALDH(high) cells and reduces ALDH(high) cell chemoresistance, (8) ALDH(high) cells are highly efficient at initiating tumors in orthotopic xenografts, and (9) AHR knockdown inhibits tumor initiation and reduces tumor Aldh1a1, Sox2, and Cyp1b1 expression in vivo. Conclusions: These data suggest that the AHR plays an important role in development of cells with cancer stem cell-like qualities and that environmental AHR ligands may exacerbate breast cancer by enhancing expression of these properties.
MeSH term(s)	Breast/metabolism ; Breast/pathology ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Cell Line, Tumor ; Cell Movement ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplastic Stem Cells/metabolism ; Neoplastic Stem Cells/pathology ; Receptors, Aryl Hydrocarbon/genetics ; Receptors, Aryl Hydrocarbon/metabolism
Chemical Substances	Receptors, Aryl Hydrocarbon
Language	English
Publishing date	2016-03-16
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2133020-7
ISSN	1741-7007 ; 1741-7007
ISSN (online)	1741-7007
ISSN	1741-7007
DOI	10.1186/s12915-016-0240-y
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Article ; Online: Role for the Aryl Hydrocarbon Receptor and Diverse Ligands in Oral Squamous Cell Carcinoma Migration and Tumorigenesis.

Stanford, Elizabeth A / Ramirez-Cardenas, Alejandra / Wang, Zhongyan / Novikov, Olga / Alamoud, Khalid / Koutrakis, Petros / Mizgerd, Joseph P / Genco, Caroline A / Kukuruzinska, Maria / Monti, Stefano / Bais, Manish V / Sherr, David H

Molecular cancer research : MCR

2016 Volume 14, Issue 8, Page(s) 696–706

Abstract: Unlabelled: Over 45,000 new cases of oral and pharyngeal cancers are diagnosed and account for over 8,000 deaths a year in the United States. An environmental chemical receptor, the aryl hydrocarbon receptor (AhR), has previously been implicated in oral ...

Abstract	Unlabelled: Over 45,000 new cases of oral and pharyngeal cancers are diagnosed and account for over 8,000 deaths a year in the United States. An environmental chemical receptor, the aryl hydrocarbon receptor (AhR), has previously been implicated in oral squamous cell carcinoma (OSCC) initiation as well as in normal tissue-specific stem cell self-renewal. These previous studies inspired the hypothesis that the AhR plays a role in both the acquisition and progression of OSCC, as well as in the formation and maintenance of cancer stem-like cells. To test this hypothesis, AhR activity in two oral squamous cell lines was modulated with AhR prototypic, environmental and bacterial AhR ligands, AhR-specific inhibitors, and phenotypic, genomic and functional characteristics were evaluated. The data demonstrate that: (i) primary OSCC tissue expresses elevated levels of nuclear AhR as compared with normal tissue, (ii) AhR mRNA expression is upregulated in 320 primary OSCCs, (iii) AhR hyperactivation with several ligands, including environmental and bacterial ligands, significantly increases AhR activity, ALDH1 activity, and accelerates cell migration, (iv) AhR inhibition blocks the rapid migration of OSCC cells and reduces cell chemoresistance, (v) AhR knockdown inhibits tumorsphere formation in low adherence conditions, and (vi) AhR knockdown inhibits tumor growth and increases overall survival in vivo These data demonstrate that the AhR plays an important role in development and progression of OSCC, and specifically cancer stem-like cells. Prototypic, environmental, and bacterial AhR ligands may exacerbate OSCC by enhancing expression of these properties. Implications: This study, for the first time, demonstrates the ability of diverse AhR ligands to regulate AhR activity in oral squamous cell carcinoma cells, as well as regulate several important characteristics of oral cancer stem cells, in vivo and in vitro Mol Cancer Res; 14(8); 696-706. ©2016 AACR.
MeSH term(s)	Animals ; Carcinogenesis ; Carcinoma, Squamous Cell/genetics ; Carcinoma, Squamous Cell/pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Disease Models, Animal ; Female ; Humans ; Ligands ; Mice ; Mice, Nude ; Mouth Neoplasms/genetics ; Mouth Neoplasms/pathology ; Receptors, Aryl Hydrocarbon/metabolism ; Signal Transduction ; Transfection
Chemical Substances	Ligands ; Receptors, Aryl Hydrocarbon
Language	English
Publishing date	2016-04-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2098788-2
ISSN	1557-3125 ; 1541-7786
ISSN (online)	1557-3125
ISSN	1541-7786
DOI	10.1158/1541-7786.MCR-16-0069
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Article ; Online: Response biases in preschool children's ratings of pain in hypothetical situations.

von Baeyer, Carl L / Forsyth, Sasha J / Stanford, Elizabeth A / Watson, Mark / Chambers, Christine T

European journal of pain (London, England)

2009 Volume 13, Issue 2, Page(s) 209–213

Abstract: Response biases are systematic biases in responding to test items that are unrelated to the content of the items. Examples often reported in young children include choosing only the lowest or highest anchors of a scale, or choosing a left-to-right ... ...

Abstract	Response biases are systematic biases in responding to test items that are unrelated to the content of the items. Examples often reported in young children include choosing only the lowest or highest anchors of a scale, or choosing a left-to-right sequence of responses. We investigated the presence of response biases in young children's ratings of pain in hypothetical situations, as a way of gauging their developing understanding of a pain scale over the preschool years. Children aged 3-5 years (N=185) rated items from the Charleston Pediatric Pain Pictures (CPPP) using the Faces Pain Scale-Revised (FPS-R). Response biases were identified objectively by computer pattern identification. Anchor biases (choosing the lowest and highest pain faces) occurred in 16% of children. Left-right or right-left sequences occurred in 35%. Monte Carlo simulation established that such patterns occur infrequently by chance (<3% for anchor biases; <6% for sequence biases). Response biases were identified more often in younger than older children. These results reveal that response biases are common in children under 5 years. Clinicians should consider self-report pain ratings from preschoolers with caution, seek complementary observational assessment, and investigate discrepancies between self-report and observational estimates of pain. Simplified forms, instructions, and methods of administration for self-report scales should be developed and validated for use with 3- and 4-year-olds.
MeSH term(s)	Aging/psychology ; Bias ; Child, Preschool ; Computer Simulation ; Data Interpretation, Statistical ; Facial Expression ; Female ; Humans ; Male ; Monte Carlo Method ; Pain Measurement/psychology ; Photic Stimulation
Language	English
Publishing date	2009-02
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1390424-3
ISSN	1532-2149 ; 1090-3801
ISSN (online)	1532-2149
ISSN	1090-3801
DOI	10.1016/j.ejpain.2008.03.017
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Article ; Online: In silico identification of an aryl hydrocarbon receptor antagonist with biological activity in vitro and in vivo.

Parks, Ashley J / Pollastri, Michael P / Hahn, Mark E / Stanford, Elizabeth A / Novikov, Olga / Franks, Diana G / Haigh, Sarah E / Narasimhan, Supraja / Ashton, Trent D / Hopper, Timothy G / Kozakov, Dmytro / Beglov, Dimitri / Vajda, Sandor / Schlezinger, Jennifer J / Sherr, David H

Molecular pharmacology

2014 Volume 86, Issue 5, Page(s) 593–608

Abstract: The aryl hydrocarbon receptor (AHR) is critically involved in several physiologic processes, including cancer progression and multiple immune system activities. We, and others, have hypothesized that AHR modulators represent an important new class of ... ...

Abstract	The aryl hydrocarbon receptor (AHR) is critically involved in several physiologic processes, including cancer progression and multiple immune system activities. We, and others, have hypothesized that AHR modulators represent an important new class of targeted therapeutics. Here, ligand shape-based virtual modeling techniques were used to identify novel AHR ligands on the basis of previously identified chemotypes. Four structurally unique compounds were identified. One lead compound, 2-((2-(5-bromofuran-2-yl)-4-oxo-4H-chromen-3-yl)oxy)acetamide (CB7993113), was further tested for its ability to block three AHR-dependent biologic activities: triple-negative breast cancer cell invasion or migration in vitro and AHR ligand-induced bone marrow toxicity in vivo. CB7993113 directly bound both murine and human AHR and inhibited polycyclic aromatic hydrocarbon (PAH)- and TCDD-induced reporter activity by 75% and 90% respectively. A novel homology model, comprehensive agonist and inhibitor titration experiments, and AHR localization studies were consistent with competitive antagonism and blockade of nuclear translocation as the primary mechanism of action. CB7993113 (IC50 3.3 × 10(-7) M) effectively reduced invasion of human breast cancer cells in three-dimensional cultures and blocked tumor cell migration in two-dimensional cultures without significantly affecting cell viability or proliferation. Finally, CB7993113 effectively inhibited the bone marrow ablative effects of 7,12-dimethylbenz[a]anthracene in vivo, demonstrating drug absorption and tissue distribution leading to pharmacological efficacy. These experiments suggest that AHR antagonists such as CB7993113 may represent a new class of targeted therapeutics for immunomodulation and/or cancer therapy.
MeSH term(s)	Animals ; Biological Factors/pharmacology ; Bone Marrow Cells/drug effects ; COS Cells ; Cell Line ; Cell Line, Tumor ; Cell Movement/drug effects ; Cell Proliferation/drug effects ; Chlorocebus aethiops ; Humans ; Ligands ; Male ; Mice ; Mice, Inbred C57BL ; Receptors, Aryl Hydrocarbon/antagonists & inhibitors ; Stromal Cells/drug effects ; Triple Negative Breast Neoplasms/drug therapy
Chemical Substances	Biological Factors ; Ligands ; Receptors, Aryl Hydrocarbon
Language	English
Publishing date	2014-08-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	124034-1
ISSN	1521-0111 ; 0026-895X
ISSN (online)	1521-0111
ISSN	0026-895X
DOI	10.1124/mol.114.093369
Database	MEDical Literature Analysis and Retrieval System OnLINE

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