Modeling statin myopathy in a human skeletal muscle microphysiological system

Statins are used to lower cholesterol and prevent cardiovascular disease. Musculoskeletal side effects known as statin associated musculoskeletal symptoms (SAMS), are reported in up to 10% of statin users, necessitating statin therapy interruption and increasing cardiovascular disease risk. We tested the hypothesis that, when exposed to statins ex vivo, engineered human skeletal myobundles derived from individuals with (n = 10) or without (n = 14) SAMS and elevated creatine-kinase levels exhibit statin-dependent muscle defects. Myoblasts were derived from muscle biopsies of individuals (median age range of 62–64) with hyperlipidemia with (n = 10) or without (n = 14) SAMS. Myobundles formed from myoblasts were cultured with growth media for 4 days, low amino acid differentiation media for 4 days, then dosed with 0 and 5μM of statins for 5 days. Tetanus forces were subsequently measured. To model the change of tetanus forces among clinical covariates, a mixed effect model with fixed effects being donor type, statin concentration, statin type and their two way interactions (donor type*statin concentration and donor type* statin type) and the random effect being subject ID was applied. The results indicate that statin exposure significantly contributed to decrease in force (P<0.001) and the variability in data (R2C [R square conditional] = 0.62). We found no significant differences in force between myobundles from patients with/without SAMS, many of whom had chronic diseases. Immunofluorescence quantification revealed a positive correlation between the number of straited muscle fibers and tetanus force (R2 = 0.81,P = 0.015) and negative correlation between number of fragmented muscle fibers and tetanus force (R2 = 0.482,P = 0.051) with no differences between donors with or without SAMS. There is also a correlation between statin exposure and presence of striated fibers (R2 = 0.833, P = 0.047). In patient-derived myobundles, statin exposure results in myotoxicity disrupting SAA organization and reducing force. We were unable to identify differences in ex vivo statin myotoxicity in this system. The results suggest that it is unlikely that there is inherent susceptibility to or persistent effects of statin myopathy using patient-derived myobundles.

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Abstract P101: Methylome-wide Association Study Provides Evidence of Particulate Matter Air Pollution-associated Dna Methylation at Cardiovascular Disease-related Genes

Circulation ◽

10.1161/circ.137.suppl_1.p101 ◽

2018 ◽

Vol 137 (suppl_1) ◽

Author(s):

Rahul Gondalia ◽

Antoine R Baldassari ◽

Katelyn M Holliday ◽

Raúl Méndez-Giráldez ◽

Anne E Justice ◽

...

Keyword(s):

Cardiovascular Disease ◽

Dna Methylation ◽

Air Pollution ◽

Particulate Matter ◽

Association Study ◽

Fixed Effects ◽

Disease Risk ◽

Epigenetic Modification ◽

Cardiovascular Disease Risk ◽

Cpg Sites

Background: DNA methylation (DNAm), a heritable but dynamic epigenetic modification that can influence gene expression without altering the genome, may underlie the associations between air pollution and cardiovascular disease risk. Therefore, our objective was to evaluate associations between DNAm and ambient concentrations of particulate matter (PM) ≤ 2.5 and ≤ 10 micrometers in diameter (PM 2.5; PM 10 ). Methods: We conducted a methylome-wide association study among twelve cohort- and race/ethnicity- stratified subpopulations from the Women’s Health Initiative and the Atherosclerosis Risk in Communities study (discovery n = 6,720; replication n = 1,936; mean age: 61.3 yrs; 83% female; 46% African American; 9% Hispanic/Latino American). We averaged geocoded address-specific estimates of daily and monthly mean PM concentrations over 2, 7, 28, and 365 days and 1 and 12 months before exams at which we measured leukocyte DNAm in whole blood. In each subpopulation, we estimated PM-DNAm associations at approximately 485,000 Cytosine-phosphate-Guanine (CpG) sites in multi-level, linear mixed-effects models adjusting for sociodemographic, behavioral, and meteorological characteristics; estimated leukocyte proportions; and technical covariates. We combined subpopulation-specific PM-DNAm associations in fixed-effects, inverse variance-weighted meta-analyses of the discovery, replication, and overall populations. Then we conducted in silico characterization of CpG sites at which PM-DNAm associations exceeded methylome-wide significance and were not heterogeneous ( P < 1.0 x 10 -7 ; P Cochran’s Q > 0.10) to assess their putative function and biological plausibility. Results: Discovery analyses identified significant PM 2.5 - and PM 10 -DNAm associations at four CpG sites, but none survived Bonferroni correction. Overall analyses identified significant associations at two CpG sites. On chromosome 20 near MATN4 , 28-day mean PM 10 was associated with increased DNAm at cg19004594 ( P all = 2.8 x 10 -8 ; P Cochran’s Q = 0.61). MATN4 is expressed in heart and lung tissues. It encodes Matrilin 4, a von Willebrand factor A domain-containing protein linked to cardiac remodeling. On chromosome 10 near ARPP21 , 1-month mean PM 10 was inversely associated with DNAm at cg24102420 ( P all = 4.8 x 10 -8 ; P Cochran’s Q = 0.51). ARPP21 is expressed in the brain/spinal cord and neutrophils. It encodes cAMP-regulated phosphoprotein 21, a regulator of calmodulin/calcium signaling. Conclusions: Findings from this methylome-wide association study suggest that ambient PM 10 concentrations affect DNA methylation at regions of the genome potentially related to cardiovascular disease among racially, ethnically and environmentally diverse populations of U.S. men and women. Further investigation is warranted to uncover epigenetic mechanisms of PM-associated cardiovascular traits.

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