2. Methods

2.1. Study design

The primary objective of this study was to evaluate the role of ALDH2 reactive aldehyde detoxification in endometriosis. First, ALDH2 activity, ALDH2 expression, and 4-HNE adduct formation were assessed in the endometrium of women with endometriosis (lesion and eutopic) compared to women without endometriosis (eutopic) to determine how ALDH2 activity was regulated. We then biochemically characterized an ALDH2* 2 knock-in mouse and used a preclinical endometriosis model to determine whether ALDH2 activity influences disease development. Subsequently, we incorporated the ALDH2 activator Alda-1 and behavioral assessments to determine whether increasing ALDH2 activity could prevent endometriosis development and/or alleviate pain-associated behaviors. For the animal studies, the number per group was based on previous experience with the disease model and a power analysis. Mice were randomly placed in control and treatment groups, and experimenters were blinded to conditions. All sample processing in this study was performed concurrently on experimental and control groups using identical methods. Experimenters quantifying results were blinded. Statistical tests and number of animals or replicates for each experiment are included in the figure legends.