Since this approach provides only very limited conclusions about the causes of the observed phenomena, a new strategy has emerged, the so-called "bottom-up analysis" that allows us to identify the causes of the changes in expression. This strategy consists of two main stages: (1) analysis of the promoters and enhancers of the identified DEG to determine the transcription factors (TF) involved in the process under study; (2) reconstruction of signaling paths that activate these TFs and identify the main regulators at the top of such paths.

We identified 10 gene regulators that can play the most important role in the accumulation of cholesterol. It is important to note that 9 out of 10 genes are associated with the molecular and cellular mechanisms of atherosclerosis and comorbidconditions (eg, hypertension, diabetes, metabolic syndrome). Most of the regulator genes (7 of 10) are associated with immune response and inflammation. Surprisingly, none of the 10 genes are directly related to intracellular cholesterol metabolism. Previously, we assumed that changes in the regulation of genes are the result of accumulation of lipids in arterial cells. It is likely that the opposite is true: non-intracellular accumulation of cholesterol causes a reaction of innate immunity, but the immune response due to the interaction of cells with modified LDL stimulates the accumulation of intracellular lipids.

In the future, we will test this hypothesis and investigate the role of the identified genes in atherogenesis