Ischemic heart disease is the leading cause of death in the Western countries. In case of an acute myocardial infarction, the most dramatic manifestation of the disease, reperfusion of the occluded coronary artery is inevitable. However, reperfusion partly exacerbates the injury by several mechanisms, causing so called ischemia-reperfusion (I/R) injury. Different cardioprotective drug therapies have been proposed to alleviate the injury and to improve regeneration of myocardium. For the time being, their translation into clinical use has not been succesful.
To search for novel therapeutic targets of the I/R injury, we performed screenings on HL-1 cardiomyocytes using an shRNA library, consisting of 25,000 shRNAs targeting 4,625 genes, and a drug library consisting of 694 clinically used drugs. To model I/R injury, cells were exposed to hypoxia-reoxygenation (H/R) treatment or grown in normoxia as controls. Relative shRNA enrichment and depletion in the H/R-treated cells, as compared to normoxic cells, was measured after sequencing of the shRNA-specific barcodes from the cell populations. For drug screening, cell viability was used as the read-out. These two screening approaches were combined for pathway analysis using dimensionality reduction. The most important signaling pathways and their representative members, most notably EGFR and PRKACA, were selected for further validations in two cardiomyocyte cell lines.
96
Normal
0
false
false
false
EN-GB
X-NONE
X-NONE
/* Style Definitions */
table.MsoNormalTable
{mso-style-name:”Table Normal”;
mso-tstyle-rowband-size:0;
mso-tstyle-colband-size:0;
mso-style-noshow:yes;
mso-style-priority:99;
mso-style-parent:””;
mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
mso-para-margin:0cm;
mso-para-margin-bottom:.0001pt;
mso-pagination:widow-orphan;
font-size:12.0pt;
font-family:”Calibri”,sans-serif;
mso-ascii-font-family:Calibri;
mso-ascii-theme-font:minor-latin;
mso-hansi-font-family:Calibri;
mso-hansi-theme-font:minor-latin;
mso-fareast-language:EN-US;}
By this wide screening approach, we found several promising druggable targets for the treatment of ischemia-reperfusion injury. The results could be used for repurposing of the tested drugs. On the other hand, information on drugs with detrimental effects to cardiomyocytes under hypoxia could be useful in identifying cardiotoxic drugs in patients with ischemic heart disease.
