Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of Immortalized cardiac cell lines for characterization and comparison

ABSTRACT: The aim of this study was to compare the most common immortalized cardiac cell lines (human: AC16, rat: H9C2, mouse: HL-1) to primary cultures (neonatal rat or mouse cardiomyocytes, and human induced pluripotent stem cells) and left ventricular tissues from the corresponding species. To characterize cardiac cell lines, cardiac cell lines were seeded onto plates, and their differentiation towards a more cardiac phenotype was induced on the basis of most commonly used protocols in literature. The cells were harvested either in stage of proliferation or differentiation, and left ventricular tissue from each corresponding species, and isolated neonatal primary cardiac myocytes (for mouse and rat) or human induced pluripotent stem cells were applied as references for comparison. Transcriptomic analysis was performed on all samples. Generally, the mRNA expression pattern of cardiac markers in the cell lines showed significant differences compared to corresponding tissue or primary cultures. mRNA profile of cell lines indicates poor cardiac characteristics regardless the differentiation protocol used. Limitations of these cell lines should be taken into account when these cells are used as in vitro platforms to model cardiomyocytes and cardiovascular diseases.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Homo sapiens

SUBMITTER: Zsófia Onódi

PROVIDER: E-MTAB-10720 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Extracellular vesicles (EV) are cell-secreted, nano-sized membrane particles with various molecular cargo, including metabolites, proteins and nucleic acids [1] . EVs are involved in various physiological and pathological functions, including that of the myocardium [2] , such as cardiac stress adaptation mechanisms [3] . Metabolic co-morbidities can ameliorate the innate stress response of the heart [4], [5] . Furthermore, one of the most common metabolic co-morbidities, type-II diabetes, can inhibit EV- mediated cardioprotection [6] . EVs of high-fat fed animals can also exacerbate ischemic cardiac damage and induce cell death of cardiomyocytes (CM) [7], [8] . Therefore, metabolic diseases might interfere with the effect of cardiac EVs by various mechanisms. However, as of now, little is known about the effect of hypercholesterolemia (HC) on cardiovascular EV communication. Analyzing EVs of the circulation might lead to novel diagnostic tools and to better understanding of disease conditions. As of now it is evidenced that circulating EV number is increased in metabolic diseases [9]–[11] , including familiar hypercholesterolemia [12], [13] , in which their amount is associated with the risk of cardiovascular diseases [14], [15] as well. However, to understand the biological role of EV-related changes in cardiometabolic diseases, characterization of EV composition is needed. The main molecular constituents of EVs are membrane lipids and other metabolites, intravesicular and transmembrane proteins and nucleotides. Change in any of these constituents may result in an altered biological response. Barrachina et. al. has analyzed the dysregulated protein composition of circulating EVs in obesity [16] and in a similar study, miRNA cargo of EVs from obese patients was analyzed, as well [17] , however, it has not been assessed if HC alters EV metabolome, thus, function. EVs contribute to response to various stresses in the myocardium [18] , but as of now, we do not know HC influences EV release of CMs. One of the major effects of HC is cardiac- and systemic inflammation [19] . Earlier studies showed in various disease conditions, such as angiotensin II-induced hypertrophy, that CM EVs activate monocytes, thereby contribute to inflammation [20], [21] . In contrast, in hypoxic conditions, EVs promote macrophage polarization to the reparative M2 phenotype [22], [23] . Thus, we hypothesized that CM EVs might play a role in HC-induced cardiac inflammation. In this study, we aimed to analyze how HC affects the metabolic composition of EVs and how HC modifies CM EV communication. Therefore, we have analyzed the metabolome of circulating EVs of HC- fed rats and compared the identified changes to the plasma metabolome. In addition, for the first time, we assessed how HC affects the biophysical and biochemical properties of CM EVs and we analyzed their role in immune activation.

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RNA-seq of Immortalized cardiac cell lines for characterization and comparison

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