Acute respiratory distress syndrome (ARDS) is a devastating clinical syndrome with a considerable case fatality rate (˜30%–40%). Health disparities exist with African descent (AD) subjects exhibiting greater mortality than European descent (ED) individuals. Myosin light chain kinase is encoded byMYLK, whose genetic variants are implicated in ARDS pathogenesis and may influence ARDS mortality. As baseline population-specific epigenetic changes, that is, cytosine modifications, have been observed between AD and ED individuals, epigenetic variations inMYLKmay provide insights into ARDS disparities. We compared methylation levels ofMYLKcytosine-guanine dinucleotides (CpGs) between ARDS patients and intensive care unit (ICU) controls overall and by ethnicity in a nested case–control study of 39 ARDS cases and 75 non-ARDS ICU controls. TwoMYLKCpG sites (cg03892735 and cg23344121) were differentially modified between ARDS subjects and controls (P< 0.05;q< 0.25) in a logistic regression model, where no effect modification by ethnicity or age was found. One CpG site was associated with ARDS in patients aged <58 years, cg19611163 (intron 19, 20). Two CpG sites were associated with ARDS in EDs only, gene body CpG (cg01894985, intron 2, 3) and CpG (cg16212219, intron 31, 32), with higher modification levels exhibited in ARDS subjects than controls.Cis-acting modified cytosine quantitative trait loci (mQTL) were identified using linear regression between local genetic variants and modification levels for 2 ARDS-associated CpGs (cg23344121 and cg16212219). In summary, these ARDS-associatedMYLKCpGs with effect modification by ethnicity and local mQTL suggest thatMYLKepigenetic variation and local genetic background may contribute to health disparities observed in ARDS.