Acetaldehyde is both an exogenous carcinogen and endogenous metabolite. While acetaldehyde formed after consumption of alcoholic beverages is well-known as a hepatotoxin and contributor to the developmental abnormalities associated with fetal alcohol syndrome, endogenous production of acetaldehyde was only recently linked to the genetic disease, Fanconi anemia. Acetaldehyde forms DNA adducts, including interstrand DNA crosslinks, repaired by the Fanconi anemia/BRCA pathway. Alcohol metabolism also induces mitochondrial autophagy, or mitophagy, as a cytoprotective mechanism. An entirely new function for Fanconi anemia/BRCA proteins in mitophagy was recently described, highlighting a potential role for the FA/BRCA in responding to acetaldehyde-induced cellular injury. Both Fanconi anemia and fetal alcohol syndrome are characterized by congenital anomalies, and Fanconi anemia and alcohol predispose to an overlapping collection of epithelial neoplasms. Cellular sensitivity to acetaldehyde is poorly understood, and is likely affected by endogenous production of acetaldehyde, metabolism, generation of covalent DNA and protein adducts, and repair pathways. Nave and primed embryonic stem cells represent pre- and post-implantation pluripotent populations in the developing embryo. We demonstrate that nave embryonic stem cells are strikingly tolerant of 1 mM acetaldehyde, while primed embryonic stem cells are exquisitely sensitive (in fact, our collaborator, Dr. Carol Ware, uses acetaldehyde routinely to select against any primed cells that may arise during culture of nave embryonic stem cells). In preliminary data, we show that both types of embryonic stem cell generate acetaldehyde and exhibit basal levels of acetaldehyde adducts with DNA to different degrees. Surprisingly, resistance of nave embryonic stem cells to acetaldehyde is not diminished by inhibition of acetaldehyde-metabolizing aldehyde dehydrogenases. In this proposal, we will investigate the mechanisms of differential acetaldehyde resistance in embryonic stem cells. Specifically, in Aim 1 we will utilize sensitive DNA adductomic profiling to examine the role of the Fanconi anemia/BRCA DNA repair pathway or mitochondrial DNA repair in acetaldehyde resistance.
In Aim 2, we will investigate the regulation of the mitophagic response to acetaldehyde by the FA/BRCA pathway. Acetaldehyde inhibits sirtuin 3, the mitochondrial deacetylase, most likely by depleting its NAD+ substrate, leading to a hyperacetylated mitochondrial proteome. Acetylation inhibits mitophagy. We will determine whether the FA/BRCA pathway reduces mitochondrial protein acetylation, thus enabling quality control via mitophagy. The proposed research will generate important new information for understanding cellular responses to acetaldehyde, both in the context of alcohol consumption and genetic syndromes of acetaldehyde hypersensitivity.

Public Health Relevance

Acetaldehyde is a product of ethanol metabolism and a major contributor to alcohol-induced diseases, including cirrhosis and fetal alcohol syndrome. It is also a normal byproduct of cellular metabolism. Fetal alcohol syndrome, and Fanconi anemia, a genetic disease proposed to result from failure to repair acetaldehyde-induced chemical crosslinks of DNA strands, feature developmental abnormalities and increased risk of certain cancers. Using two embryonic stem cell types with greatly different sensitivities to acetaldehyde, we will identify why certain cell types in the developing embryo are sensitive to acetaldehyde, and how the ability of cells to produce acetaldehyde during normal metabolism affects their tolerance to toxic effects of ethanol.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AA026398-02
Application #
9569574
Study Section
National Institute on Alcohol Abuse and Alcoholism Initial Review Group (AA)
Program Officer
Gao, Peter
Project Start
2017-09-22
Project End
2019-08-31
Budget Start
2018-09-01
Budget End
2019-08-31
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109