High-Content Image-Based Single-Cell Phenotypic Analysis for the Testicular Toxicity Prediction Induced by Bisphenol A and Its Analogs Bisphenol S, Bisphenol AF, and Tetrabromobisphenol A in a Three-Dimensional Testicular Cell Co-Culture Model

Details

• Personal Author:
  Edenfield C ; Guan X ; Liang S ; Measel E ; Siracusa JS ; Yin L ; Yu X
• Description:
  Emerging data indicate that structural analogs of bisphenol A (BPA) such as bisphenol S (BPS), tetrabromobisphenol A (TBBPA), and bisphenol AF (BPAF) have been introduced into the market as substitutes for BPA. Our previous study compared in vitro testicular toxicity using murine C18-4 spermatogonial cells and found that BPAF and TBBPA exhibited higher spermatogonial toxicities as compared with BPA and BPS. Recently, we developed a novel in vitro three-dimensional (3D) testicular cell co-culture model, enabling the classification of reproductive toxic substances. In this study, we applied the testicular cell co-culture model and employed a high-content image (HCA)-based single-cell analysis to further compare the testicular toxicities of BPA and its analogs. We also developed a machine learning (ML)-based HCA pipeline to examine the complex phenotypic changes associated with testicular toxicities. We found dose- and time-dependent changes in a wide spectrum of adverse endpoints, including nuclear morphology, DNA synthesis, DNA damage, and cytoskeletal structure in a single-cell-based analysis. The co-cultured testicular cells were more sensitive than the C18 spermatogonial cells in response to BPA and its analogs. Unlike conventional population-averaged assays, single-cell-based assays not only showed the levels of the averaged population, but also revealed changes in the sub-population. Machine learning-based phenotypic analysis revealed that treatment of BPA and its analogs resulted in the loss of spatial cytoskeletal structure, and an accumulation of M phase cells in a dose- and time-dependent manner. Furthermore, treatment of BPAF-induced multinucleated cells, which were associated with altered DNA damage response and impaired cellular F-actin filaments. Overall, we demonstrated a new and effective means to evaluate multiple toxic endpoints in the testicular co-culture model through the combination of ML and high-content image-based single-cell analysis. This approach provided an in-depth analysis of the multi-dimensional HCA data and provided an unbiased quantitative analysis of the phenotypes of interest. [Description provided by NIOSH]
• Subjects:
  Occupational Health Safety Spermatogenesis Toxicology Urogenital System
• Keywords:
  Author Keywords: Bisphenol A Bisphenol A Bisphenol AF Bisphenol S Cell Culture Techniques DNA Damage High-content Image In Vitro Co-culture Model In Vitro Study Machine Learning Morphology Single-cell Analysis Testes Testicular Toxicity Tetrabromobisphenol A

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• ISSN:
  1096-6080
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  Georgia ; OSHA Region 4
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  313-335
• Volume:
  173
• Issue:
  2
• NIOSHTIC Number:
  nn:20068434
• Citation:
  Toxicol Sci 2020 Feb; 173(2):313-335
• Contact Point Address:
  Xiaozhong Yu, Department of Environmental Health Science, College of Public Health, University of Georgia, Athens, GA 30602
• Email:
  yuxz@uga.edu
• CAS Registry Number:
  2,2-Bis(4-hydroxyphenyl)propane (CAS RN 80-05-7)
• Federal Fiscal Year:
  2020
• Performing Organization:
  University of Georgia, Athens
• Peer Reviewed:
  True
• Start Date:
  20130901
• Source Full Name:
  Toxicological Sciences
• End Date:
  20160831
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:485fe613e7c8dfb9b2e0c835ba532c24ce680f8069a9b07f384831e4201f0f4da72e41ca189b970399c34c5d8051d9c2109e4dd0c2f1a1552cbb7f5e7d0c03a7
• Download URL:
  https://stacks.cdc.gov/view/cdc/230589/cdc_230589_DS1.pdf
• File Type:
  [PDF - 3.30 MB ]