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Article ; Online: Pluripotent stem cells for target organ developmental toxicity testing.

Wu, Xian / Chen, Yichang / Kreutz, Anna / Silver, Brian / Tokar, Erik J

Toxicological sciences : an official journal of the Society of Toxicology

2024

Abstract: Prenatal developmental toxicity research focuses on understanding potential adverse effects of environmental agents, drugs, and chemicals on the development of embryos and fetuses. Traditional methods involve animal testing, but ethical concerns and the ... ...

Abstract	Prenatal developmental toxicity research focuses on understanding potential adverse effects of environmental agents, drugs, and chemicals on the development of embryos and fetuses. Traditional methods involve animal testing, but ethical concerns and the need for human-relevant models have prompted the exploration of alternatives. Pluripotent stem cells (PSCs) are versatile cells with the unique ability to differentiate into any cell type, serving as a foundational tool for studying human development. Two-dimensional (2D) PSC models are often chosen for their ease of use and reproducibility for high-throughput screening. However, they lack the complexity of an in vivo environment. Alternatively, three-dimensional (3D) PSC models, such as organoids, offer tissue architecture and intercellular communication more reminiscent of in vivo conditions. However, they are complicated to produce and analyze, usually requiring advanced and expensive techniques. This review discusses recent advances in the use of human PSCs differentiated into brain and heart lineages and emerging tools and methods that can be combined with PSCs to help address important scientific questions in the area developmental toxicology. These advancements and new approach methods align with the push for more relevant and predictive developmental toxicity assessment, combining innovative techniques with organoid models to advance regulatory decision-making.
Language	English
Publishing date	2024-03-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	1420885-4
ISSN	1096-0929 ; 1096-6080
ISSN (online)	1096-0929
ISSN	1096-6080
DOI	10.1093/toxsci/kfae037
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Article ; Online: Molecular Mechanisms of Cytotoxicity of NCX4040, the Non-Steroidal Anti-Inflammatory NO-Donor, in Human Ovarian Cancer Cells.

Sinha, Birandra K / Tokar, Erik J / Bortner, Carl D

International journal of molecular sciences

2022 Volume 23, Issue 15

Abstract: NCX4040, the non-steroidal anti-inflammatory-NO donor, is cytotoxic to several human tumors, including ovarian tumor cells. We have found that NCX4040 is also cytotoxic against both OVCAR-8 and its adriamycin resistant (NCI/ADR-RES) tumor cell lines. ... ...

Abstract	NCX4040, the non-steroidal anti-inflammatory-NO donor, is cytotoxic to several human tumors, including ovarian tumor cells. We have found that NCX4040 is also cytotoxic against both OVCAR-8 and its adriamycin resistant (NCI/ADR-RES) tumor cell lines. Here, we have examined mechanism(s) for the cytotoxicity of NCX4040 in OVCAR-8 and NCI/ADR-RES cell lines. We found that NCX4040 induced significant apoptosis in both cell lines. Furthermore, NCX4040 treatment caused significant depletion of cellular glutathione, causing oxidative stress due to the formation of reactive oxygen/nitrogen species (ROS/RNS). Significantly more ROS/RNS were detected in OVCAR-8 cells than in NCI/ADR-RES cells which may have resulted from increased activities of SOD, glutathione peroxidase and transferases expressed in NCI/ADR-RES cells. NCX4040 treatment resulted in the formation of double-strand DNA breaks in both cells; however, more of these DNA breaks were detected in OVCAR-8 cells. RT-PCR studies indicated that NCX4040-induced DNA damage was not repaired as efficiently in NCI/ADR-RES cells as in OVCAR-8 cells which may lead to a differential cell death. Pretreatment of OVCAR-8 cells with N-acetylcysteine (NAC) significantly decreased cytotoxicity of NCX4040 in OVCAR-8 cells; however, NAC had no effects on NCX4040 cytotoxicity in NCI/ADR-RES cells. In contrast, FeTPPS, a peroxynitrite scavenger, completely blocked NCX4040-induced cell death in both cells, suggesting that NCX4040-induced cell death could be mediated by peroxynitrite formed from NCX4040 following cellular metabolism.
MeSH term(s)	Anti-Inflammatory Agents, Non-Steroidal/therapeutic use ; Antineoplastic Agents/pharmacology ; Aspirin/analogs & derivatives ; Carcinoma, Ovarian Epithelial ; Doxorubicin/pharmacology ; Female ; Humans ; Nitro Compounds ; Ovarian Neoplasms/pathology ; Peroxynitrous Acid ; Reactive Nitrogen Species ; Reactive Oxygen Species
Chemical Substances	Anti-Inflammatory Agents, Non-Steroidal ; Antineoplastic Agents ; NCX 4040 ; Nitro Compounds ; Reactive Nitrogen Species ; Reactive Oxygen Species ; Peroxynitrous Acid (14691-52-2) ; Doxorubicin (80168379AG) ; Aspirin (R16CO5Y76E)
Language	English
Publishing date	2022-08-03
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms23158611
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Article ; Online: MicroRNAs: small molecules with big effects.

Tokar, Erik J

Toxicology and applied pharmacology

2016 Volume 312, Page(s) 1–2

MeSH term(s)	Animals ; Biomarkers/metabolism ; Humans ; MicroRNAs/biosynthesis ; MicroRNAs/genetics ; Neoplasms/genetics ; Neoplasms/metabolism
Chemical Substances	Biomarkers ; MicroRNAs
Language	English
Publishing date	2016--01
Publishing country	United States
Document type	Editorial
ZDB-ID	204477-8
ISSN	1096-0333 ; 0041-008X
ISSN (online)	1096-0333
ISSN	0041-008X
DOI	10.1016/j.taap.2016.11.004
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Article ; Online: Stem Cells as Target for Prostate cancer Therapy: Opportunities and Challenges.

Escudero-Lourdes, Claudia / Alvarado-Morales, Ildemar / Tokar, Erik J

Stem cell reviews and reports

2022 Volume 18, Issue 8, Page(s) 2833–2851

Abstract: Cancer stem cells (CSCs) and cells in a cancer stem cell-like (CSCL) state have proven to be responsible for tumor initiation, growth, and relapse in Prostate Cancer (PCa) and other cancers; therefore, new strategies are being developed to target such ... ...

Abstract	Cancer stem cells (CSCs) and cells in a cancer stem cell-like (CSCL) state have proven to be responsible for tumor initiation, growth, and relapse in Prostate Cancer (PCa) and other cancers; therefore, new strategies are being developed to target such cellular populations. TLR3 activation-based immunotherapy using Polyinosinic:Polycytidylic acid (PIC) has been proposed to be used as a concomitant strategy to first-line treatment. This strategy is based on the induction of apoptosis and an inflammatory response in tumor cells. In combination with retinoids like 9cRA, this treatment can induce CSCs differentiation and apoptosis. A limitation in the use of this combination is the common decreased expression of TLR3 and its main positive regulator p53. observed in many patients suffering of different cancer types such as PCa. Importantly, human exposure to certain toxicants, such as iAs, not only has proven to enrich CSCs population in an in vitro model of human epithelial prostate cells, but additionally, it can also lead to a decreased p53, TLR3 and RA receptor (RARβ), expression/activation and thus hinder this treatment efficacy. Therefore, here we point out the relevance of evaluating the TLR3 and P53 status in PCa patients before starting an immunotherapy based on the use of PIC +9cRA to determine whether they will be responsive to treatment. Additionally, the use of strategies to overcome the lower TLR3, RARβ or p53 expression in PCa patients, like the inclusion of drugs that increase p53 expression, is encouraged, to potentiate the use of PIC+RA based immunotherapy in these patients.
MeSH term(s)	Male ; Humans ; Prostate/metabolism ; Prostate/pathology ; Toll-Like Receptor 3/metabolism ; Tumor Suppressor Protein p53/genetics ; Neoplasm Recurrence, Local ; Prostatic Neoplasms/therapy ; Prostatic Neoplasms/drug therapy ; Stem Cells/metabolism
Chemical Substances	Toll-Like Receptor 3 ; Tumor Suppressor Protein p53
Language	English
Publishing date	2022-08-11
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	2495577-2
ISSN	2629-3277 ; 1558-6804 ; 1550-8943
ISSN (online)	2629-3277 ; 1558-6804
ISSN	1550-8943
DOI	10.1007/s12015-022-10437-6
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Article ; Online: Arsenic as an environmental toxicant and a therapeutic agent: Foe and friend.

Xu, Yuanyuan / Tokar, Erik J / Pi, Jingbo

Toxicology and applied pharmacology

2021 Volume 415, Page(s) 115438

MeSH term(s)	Animals ; Antineoplastic Agents/adverse effects ; Arsenic/adverse effects ; Arsenic Poisoning/etiology ; Arsenic Poisoning/genetics ; Arsenic Poisoning/metabolism ; Arsenic Poisoning/pathology ; Cell Transformation, Neoplastic/chemically induced ; Cell Transformation, Neoplastic/genetics ; Cell Transformation, Neoplastic/metabolism ; Cell Transformation, Neoplastic/pathology ; DNA Damage ; Epigenesis, Genetic/drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasms/chemically induced ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology ; Nervous System/drug effects ; Nervous System/metabolism ; Nervous System/pathology ; Patient Safety ; Risk Assessment ; Risk Factors
Chemical Substances	Antineoplastic Agents ; Arsenic (N712M78A8G)
Language	English
Publishing date	2021-02-04
Publishing country	United States
Document type	Editorial ; Introductory Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	204477-8
ISSN	1096-0333 ; 0041-008X
ISSN (online)	1096-0333
ISSN	0041-008X
DOI	10.1016/j.taap.2021.115438
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Article ; Online: Pluripotent Stem Cells in Developmental Toxicity Testing: A Review of Methodological Advances.

Luz, Anthony L / Tokar, Erik J

Toxicological sciences : an official journal of the Society of Toxicology

2018 Volume 165, Issue 1, Page(s) 31–39

Abstract: Millions of children are born each year with a birth defect. Many of these defects are caused by environmental factors, although the underlying etiology is often unknown. In vivo mammalian models are frequently used to determine if a chemical poses a ... ...

Abstract	Millions of children are born each year with a birth defect. Many of these defects are caused by environmental factors, although the underlying etiology is often unknown. In vivo mammalian models are frequently used to determine if a chemical poses a risk to the developing fetus. However, there are over 80 000 chemicals registered for use in the United States, many of which have undergone little safety testing, necessitating the need for higher-throughput methods to assess developmental toxicity. Pluripotent stem cells (PSCs) are an ideal in vitro model to investigate developmental toxicity as they possess the capacity to differentiate into nearly any cell type in the human body. Indeed, a burst of research has occurred in the field of stem cell toxicology over the past decade, which has resulted in numerous methodological advances that utilize both mouse and human PSCs, as well as cutting-edge technology in the fields of metabolomics, transcriptomics, transgenics, and high-throughput imaging. Here, we review the wide array of approaches used to detect developmental toxicants, suggest areas for further research, and highlight critical aspects of stem cell biology that should be considered when utilizing PSCs in developmental toxicity testing.
MeSH term(s)	Animal Testing Alternatives ; Animals ; Cell Culture Techniques ; Cell Differentiation/drug effects ; Cell Survival/drug effects ; Embryonic Development/drug effects ; Embryonic Stem Cells/drug effects ; Embryonic Stem Cells/pathology ; Humans ; Induced Pluripotent Stem Cells/drug effects ; Induced Pluripotent Stem Cells/pathology ; Models, Biological ; Myocytes, Cardiac/drug effects ; Myocytes, Cardiac/pathology ; Toxicity Tests/methods
Language	English
Publishing date	2018-08-31
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	1420885-4
ISSN	1096-0929 ; 1096-6080
ISSN (online)	1096-0929
ISSN	1096-6080
DOI	10.1093/toxsci/kfy174
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Article: Elucidation of Mechanisms of Topotecan-Induced Cell Death in Human Breast MCF-7 Cancer Cells by Gene Expression Analysis.

Sinha, Birandra K / Tokar, Erik J / Bushel, Pierre R

Frontiers in genetics

2020 Volume 11, Page(s) 775

Abstract: Topotecan is a clinically active anticancer agent for the management of various human tumors. While the principal mechanism of tumor cell killing by topotecan is due to its interactions with topoisomerase I and formation of DNA double-strand breaks, ... ...

Abstract	Topotecan is a clinically active anticancer agent for the management of various human tumors. While the principal mechanism of tumor cell killing by topotecan is due to its interactions with topoisomerase I and formation of DNA double-strand breaks, recent studies suggest that mechanisms involving generation of reactive free radicals and induction of oxidative stress may play a significant role in topotecan-dependent tumor cell death. We have shown that topotecan generates a topotecan radical following one-electron oxidation by a peroxidase-hydrogen peroxide system which reacts with reduced glutathione and cysteine, forming the glutathiyl and cysteinyl radicals, respectively. While little is known how these events are involved in topotecan-induced tumor cell death, we have now examined the effects of topotecan short (1 h) and long (24 h) exposure on global gene expression patterns using gene expression microarray analysis in human breast MCF-7 cancer cells, a wild-type p53 containing cell line. We show here that topotecan treatment significantly down-regulated estrogen receptor alpha (ERα/ESR1) and antiapoptotic BCL2 genes in addition to many other p53-regulated genes. Furthermore, 8-oxoguanine DNA glycosylase (OGG1), ferredoxin reductase (FDXR), methionine sulfoxide reductase (MSR), glutathione peroxidases (GPx), and glutathione reductase (GSR) genes were also differentially expressed by topotecan treatment. The differential expression of these genes was observed in a wild-type p53-containing breast ZR-75-1 tumor cell line following topotecan treatment. The involvement of reactive oxygen free radical sensor genes, the oxidative DNA damage (OGG1) repair gene and induction of pro-apoptotic genes suggest that reactive free radical species play a role in topotecan-induced tumor cell death.
Language	English
Publishing date	2020-07-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2020.00775
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Article ; Online: Cardiac Development in the Presence of Cadmium: An

Wu, Xian / Chen, Yichang / Luz, Anthony / Hu, Guang / Tokar, Erik J

Environmental health perspectives

2022 Volume 130, Issue 11, Page(s) 117002

Abstract: Background: Exposure to cadmium (Cd) is associated with cardiovascular diseases. Maternal Cd exposure is a significant risk factor for congenital heart disease. However, mechanisms of Cd on developmental cardiotoxicity are not well defined.: ... ...

Abstract	Background: Exposure to cadmium (Cd) is associated with cardiovascular diseases. Maternal Cd exposure is a significant risk factor for congenital heart disease. However, mechanisms of Cd on developmental cardiotoxicity are not well defined. Objectives: We evaluated the effects of Cd on the different stages (mesoderm, cardiac induction, cardiac function) of cardiac development using an early embryo development Methods: Embryonic stem cells (ESCs) form 3D aggregates, called embryoid bodies, that recapitulate events involved with early embryogenesis (e.g., germ layer formation). This model was used for early germ layer formation and signaling pathway identification. The 2D cardiomyocyte differentiation from the Results: Cd ( Discussion: In conclusion, using a human ESC-derived 2D/3D
MeSH term(s)	Humans ; Human Embryonic Stem Cells ; Cadmium/toxicity ; Cadmium/metabolism ; Organoids ; Cell Differentiation ; Myocytes, Cardiac/metabolism ; Transcription Factors/metabolism
Chemical Substances	Cadmium (00BH33GNGH) ; Transcription Factors
Language	English
Publishing date	2022-11-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	195189-0
ISSN	1552-9924 ; 0091-6765 ; 1078-0475
ISSN (online)	1552-9924
ISSN	0091-6765 ; 1078-0475
DOI	10.1289/EHP11208
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Article ; Online: Ferroptosis-Mediated Cell Death Induced by NCX4040, The Non-Steroidal Nitric Oxide Donor, in Human Colorectal Cancer Cells: Implications in Therapy.

Sinha, Birandra K / Bortner, Carl D / Jarmusch, Alan K / Tokar, Erik J / Murphy, Carri / Wu, Xian / Winter, Heather / Cannon, Ronald E

Cells

2023 Volume 12, Issue 12

Abstract: Our recent studies show that the treatment of human ovarian tumor cells with NCX4040 results in significant depletions of cellular glutathione, the formation of reactive oxygen/nitrogen species and cell death. NCX4040 is also cytotoxic to several human ... ...

Abstract	Our recent studies show that the treatment of human ovarian tumor cells with NCX4040 results in significant depletions of cellular glutathione, the formation of reactive oxygen/nitrogen species and cell death. NCX4040 is also cytotoxic to several human colorectal cancer (CRC) cells in vitro and in vivo. Here, we examined the ferroptosis-dependent mechanism(s) of cytotoxicity of NCX4040 in HT-29 and K-RAS mutant HCT 116 colon cell lines. Ferroptosis is characterized by the accumulation of reactive oxygen species (ROS) within the cell, leading to an iron-dependent oxidative stress-mediated cell death. However, its relevance in the mechanism of NCX4040 cytotoxicity in CRCs is not known. We found that NCX4040 generates ROS in CRC cells without any depletion of cellular GSH. Combinations of NCX4040 with erastin (ER) or RSL3 (RAS-selective lethal 3), known inducers of ferroptosis, enhanced CRC death. In contrast, ferrostatin-1, an inhibitor of ferroptosis, significantly inhibited NCX4040-induced cell death. Treatment of CRC cells with NCX4040 resulted in the induction of lipid peroxidation in a dose- and time-dependent manner. NCX4040 treatment induced several genes related to ferroptosis (e.g., CHAC1, GPX4 and NOX4) in both cell lines. Metabolomic studies also indicated significant increases in both lipid and energy metabolism following the drug treatment in HT-29 and HCT 116 cells. These observations strongly suggest that NCX4040 causes the ferroptosis-mediated cell death of CRC cells. Furthermore, combinations of NCX4040 and ER or RSL3 may contribute significantly to the treatment of CRC, including those that are difficult to treat due to the presence of Ras mutations in the clinic. NCX4040-induced ferroptosis may also be a dynamic form of cell death for the treatment of other cancers.
MeSH term(s)	Humans ; Nitric Oxide Donors/pharmacology ; Ferroptosis ; Reactive Oxygen Species/metabolism ; Carbolines/pharmacology ; Cell Death ; Glutathione/metabolism ; Colorectal Neoplasms/drug therapy
Chemical Substances	NCX 4040 ; Nitric Oxide Donors ; Reactive Oxygen Species ; Carbolines ; Glutathione (GAN16C9B8O)
Language	English
Publishing date	2023-06-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12121626
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Article ; Online: Cadmium Exposure and Ovarian Reserve in Women Aged 35-49 Years: The Impact on Results From the Creatinine Adjustment Approach Used to Correct for Urinary Dilution.

Upson, Kristen / O'Brien, Katie M / Hall, Janet E / Tokar, Erik J / Baird, Donna D

American journal of epidemiology

2020 Volume 190, Issue 1, Page(s) 116–124

Abstract: Cadmium is toxic to the ovaries in animal studies, but its association with diminished ovarian reserve in women is not established. We investigated urinary cadmium, a biomarker of long-term exposure, in relation to diminished ovarian reserve, as ... ...

Abstract	Cadmium is toxic to the ovaries in animal studies, but its association with diminished ovarian reserve in women is not established. We investigated urinary cadmium, a biomarker of long-term exposure, in relation to diminished ovarian reserve, as indicated by elevated serum follicle-stimulating hormone concentrations (≥10 IU/L), in women aged 35-49 years (unweighted n = 1,681). Using data from the Third National Health and Nutrition Examination Survey (1988-1994), we conducted Poisson regression to estimate adjusted relative risks and 95% confidence intervals. Because the best approach to correcting for urinary dilution in spot samples with creatinine remains controversial, we employed 3 approaches: standardization, covariate adjustment, and covariate-adjusted standardization. Our data suggested a modest association with standardization (highest quartile vs. lowest: relative risk (RR) = 1.3, 95% confidence interval (CI): 0.8, 1.9; P for trend = 0.06) and covariate-adjusted standardization (highest quartile vs. lowest: RR = 1.3, 95% CI: 0.9, 1.9; P for trend = 0.05) and a stronger association with covariate adjustment (highest quartile vs. lowest: RR = 1.8, 95% CI: 1.2, 2.9; P for trend = 0.01). The stronger association with covariate adjustment may reflect bias from conditioning on urinary creatinine, a collider in the hypothesized causal pathway. We conclude that cadmium may contribute to ovarian aging in women and that careful consideration of the creatinine adjustment approach is needed to minimize bias.
MeSH term(s)	Adult ; Biomarkers/blood ; Biomarkers/urine ; Cadmium/urine ; Creatinine/urine ; Environmental Exposure ; Female ; Follicle Stimulating Hormone/blood ; Humans ; Middle Aged ; Nutrition Surveys ; Ovarian Reserve ; United States ; Urinalysis
Chemical Substances	Biomarkers ; Cadmium (00BH33GNGH) ; Follicle Stimulating Hormone (9002-68-0) ; Creatinine (AYI8EX34EU)
Language	English
Publishing date	2020-04-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural
ZDB-ID	2937-3
ISSN	1476-6256 ; 0002-9262
ISSN (online)	1476-6256
ISSN	0002-9262
DOI	10.1093/aje/kwaa037
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