Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)2022-11-152022-11-152022-01-11Cushen, S. C., Ricci, C. A., Bradshaw, J. L., Silzer, T., Blessing, A., Sun, J., Zhou, Z., Scroggins, S. M., Santillan, M. K., Santillan, D. A., Phillips, N. R., & Goulopoulou, S. (2022). Reduced Maternal Circulating Cell-Free Mitochondrial DNA Is Associated With the Development of Preeclampsia. Journal of the American Heart Association, 11(2), e021726. https://doi.org/10.1161/JAHA.121.0217262047-9980https://hdl.handle.net/20.500.12503/31924Background Circulating cell-free mitochondrial DNA (ccf-mtDNA) is a damage-associated molecular pattern that reflects cell stress responses and tissue damage, but little is known about ccf-mtDNA in preeclampsia. The main objectives of this study were to determine (1) absolute concentrations of ccf-mtDNA in plasma and mitochondrial DNA content in peripheral blood mononuclear cells and (2) forms of ccf-mtDNA transport in blood from women with preeclampsia and healthy controls. In addition, we sought to establish the association between aberrance in circulating DNA-related metrics, including ccf-mtDNA and DNA clearance mechanisms, and the clinical diagnosis of preeclampsia using bootstrapped penalized logistic regression. Methods and Results Absolute concentrations of ccf-mtDNA were reduced in plasma from women with preeclampsia compared with healthy controls (P</=0.02), while mtDNA copy number in peripheral blood mononuclear cells did not differ between groups (P>0.05). While the pattern of reduced ccf-mtDNA in patients with preeclampsia remained, DNA isolation from plasma using membrane lysis buffer resulted in 1000-fold higher ccf-mtDNA concentrations in the preeclampsia group (P=0.0014) and 430-fold higher ccf-mtDNA concentrations in the control group (P<0.0001). Plasma from women with preeclampsia did not induce greater Toll-like receptor-9-induced nuclear factor kappa-light-chain enhancer of activated B cells-dependent responses in human embryonic kidney 293 cells overexpressing the human TLR-9 gene (P>0.05). Penalized regression analysis showed that women with preeclampsia were more likely to have lower concentrations of ccf-mtDNA as well as higher concentrations of nuclear DNA and DNase I compared with their matched controls. Conclusions Women with preeclampsia have aberrant circulating DNA dynamics, including reduced ccf-mtDNA concentrations and DNA clearance mechanisms, compared with gestational age-matched healthy pregnant women.http://creativecommons.org/licenses/by-nc-nd/4.0/DNase Icell-free DNAmitochondrial DNApenalized regression analysispreeclampsiaCell-Free Nucleic Acids / geneticsDNA, Mitochondrial / geneticsFemaleHumansLeukocytes, MononuclearMitochondria / geneticsPre-Eclampsia / diagnosisPre-Eclampsia / geneticsPregnancyArticle© 2022 The Authors.112