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Enzymatic Acrolein Production System and Its Impact on Human Cells.
Katherine A Hurley1, Jacob Folz1, Jasmin Zgraggen1
1Department of Health Sciences and Technology, ETH Zürich, Zurich 8092, Switzerland.
Chemical Research in Toxicology
|August 19, 2024
Summary
Researchers developed a novel system to continuously produce acrolein in cell cultures. This method overcomes acrolein
Area of Science:
- Environmental toxicology
- Cell biology
- Biochemistry
Background:
- Acrolein is an environmental toxicant and gut microbe metabolite.
- Acrolein's instability in cell culture media limits biological impact studies.
- Existing methods hinder controlled acrolein exposure research.
Purpose of the Study:
- To develop a stable extracellular acrolein biosynthesis system for human cell culture.
- To enable controlled and continuous acrolein exposure studies.
- To overcome limitations of acrolein's rapid dissipation in vitro.
Main Methods:
- Utilized spermine as a precursor with fetal calf serum's amine oxidase.
- Incorporated catalase to neutralize hydrogen peroxide byproduct.
- Calibrated system responses by comparing transcriptional, DNA adduct, and cytotoxicity data with pure acrolein exposure in human colon cells.
Main Results:
- Confirmed amine oxidase activity in calf serum and catalase's necessity.
- Observed similar upregulation of oxidative stress genes (HMOX1) and colorectal cancer-related gene (SEMA4A) compared to pure acrolein.
- Demonstrated dose-dependent formation of acrolein-DNA adducts (γ-OH-Acr-dG) with a maximum of 1065 adducts per 10^8 nucleosides.
Conclusions:
- The developed enzymatic system provides a stable and controlled method for acrolein exposure in vitro.
- This system mimics biological acrolein generation and exposure relevant to human health.
- Overcomes challenges associated with acrolein's chemical instability and limited availability for research.
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