Exploring the molecular and functional cellular response to hydrazine via transcriptomics and DNA repair mutant cell lines

Abstract

Hydrazine is a rodent carcinogen and is classified as a probable human carcinogen by IARC. Though hydrazine is positive in both in vitro and in vivo DNA strand break (comet) assays, hydrazine was reported to be negative in an in vitro mutation Muta Mouse lung epithelial cell (FE1) test, as well as in a regulatory-compliant, in vivo Big Blue mouse mutation test. In this article, mechanistic studies explored the cellular response to hydrazine. When tested in a regulatory-compliant mouse lymphoma assay, hydrazine yielded unusual, weakly positive results. This prompted an investigation into the transcriptional response to hydrazine in FE1 cells via RNA sequencing. Amongst the changes identified was a dose-dependent increase in G2/M DNA damage checkpoint activation associated genes. Flow cytometric experiments in FE1 cells revealed that hydrazine exposure led to S-phase cell cycle arrest. Clonogenic assays in a variety of cell lines harboring key DNA repair protein deficiencies indicated that hydrazine could sensitize cells lacking homology dependent repair proteins (Brca2 and Fancg). Lastly, hprt assays with hydrazine were conducted to determine whether a lack of DNA repair could lead to mutagenicity. However, no robust, dose-dependent induction of mutations was noted. The transcriptional and cell cycle response to hydrazine, coupled with functional investigations of DNA repair-deficient cell lines support the inconsistencies noted in the genetic toxicology regulatory battery. In summary, while hydrazine may be genotoxic, transcriptional and functional processes involved in cell cycle regulation and DNA repair appear to play a nuanced role in mediating the mutagenic potential.

Synthesis and mutagenic risk of avanafil's potential genotoxic impurities

“…Genotoxic impurities refer to DNA reactive substances that could directly cause DNA damage at low levels, leading to DNA mutation and even carcinogenesis. 8,9 Hydrazine has been identied as a mutagenic and carcinogenic compound, [10][11][12][13][14][15] and the hydrazine group is also an alert structure of typical genotoxic carcinogens in compounds. [16][17][18] In the synthesis of ava-nal, EDCI and HOBT were used as acid-amine condensation agents, while hydrazine was used in the upper source process of HOBT, resulting in residual hydrazine in HOBT.…”