Influence of two anti-tumor drugs, pazopanib, and axitinib, on the development and thyroid-axis of zebrafish (Danio rerio) embryos/larvae

Front Endocrinol (Lausanne). 2023 Jul 31:14:1204678. doi: 10.3389/fendo.2023.1204678. eCollection 2023.

Abstract

Introduction: In recent years, the potential toxicities of different pharmaceuticals toward the thyroid system have received increasing attention. In this study, we aim to evaluate the toxic effects of pazopanib and axitinib, two anti-tumor drugs with widespread clinical use, on thyroid function in the zebrafish model.

Methods: We measured levels of thyroid-related hormones using the commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit. Whole-mount in situ hybridization (WISH) analysis was employed to detect target gene expression changes. Morphology of the thyroid were evaluated by using transgenic Tg (tg: EGFP) fish line under a confocal microscope. The relative mRNA expression of key genes was verified through quantitative real-time polymerase chain reaction (RT‒qPCR). The size and number of the follicles was quantified whereby Hematoxylin-Eosin (H & E) staining under a light microscope.

Results: The results revealed that fertilized zebrafish embryos were incubated in pazopanib or axitinib for 96 hours, development and survival were significantly affected, which was accompanied by significant disturbances in thyroid endocrine system (e.g., increased thyroid-stimulating hormone (TSH) content and decreased triiodothyronine (T3) and thyroxine (T4) content, as well as transcription changes of genes associated with the hypothalamus-pituitary-thyroid (HPT) axis. Moreover, based on whole-mount in situ hybridization staining of tg and histopathological examination of zebrafish embryos treated with pazopanib and axitinib, we observed a significantly abnormal development of thyroid follicles in the Tg (tg: EGFP) zebrafish transgenic line.

Conclusion: Collectively, these findings indicate that pazopanib and axitinib may have toxic effects on thyroid development and function, at least partially, by influencing the regulation of the HPT axis. Thus, we believe that the potential thyroid toxicities of pazopanib and axitinib in their clinical applications should receive greater attention.

Keywords: hypothalamuspituitary-thyroid (HPT) axis; hypothyroidism; thyroid disruption; thyroid histomorphology; zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Antineoplastic Agents*
  • Axitinib
  • Larva
  • Thyroid Gland
  • Zebrafish*

Substances

  • Axitinib
  • pazopanib
  • Antineoplastic Agents

Grants and funding

This work was financially supported by Chinese National Key Research Program (2022YFA0807000, 2017YFC1001801), the National Natural Science Foundation of China (No. 82000804, 81661168016, 81870537, 81670717, 82270826 and 81870540), the National Key Research and Development Program of China (2022YFA0807000), the Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province (No. 20220208) and the Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University (No.440).