Circulating Cells in Coronary Collateral Artery Growth II

In the present study, we performed transcriptome expression analyses in three independent peripheral blood-derived monocyte subpopulations from patients with chronic coronary occlusions (CTO) and tested for arteriogenesis. Whole-genome mRNA expression analyses were performed on these three monocyte subpopulations, namely: (1) unstimulated-, (2) 3 hours LPS-stimulated-, (3) monocyte-derived macrophages. Whole-genome mRNA expression analysis amongst others confirmed increased expression of IFNbeta-regulated genes in patients with insufficient coronary collateralization (collateral flow index (CFI) ≤0.37 “nonresponders”), compared with patients having sufficient collateralization (CFI>0.37 “responders”). Mononuclear cells were collected from 50 ml of arterial blood using Ficoll separation. Subsequently, monocytes were negatively isolated using immunomagnetic beads. Thereafter, from each patient the cells were divided into three separate pools that were separately treated: (1) untreated, (2) 3 hours LPS (10 ng/mL)-treated, and (3) differentiated into macrophages by 20 hours LPS (10 ng/mL) treatment. Within each of these three cell populations the transcriptome profiles were divided into two patient groups, being the non-responders (CFI≤0.37) versus the responders (CFI>0.37). Both patient groups had been matched for age, sex, medication and other factors that influence collateral artery growth. The mRNA-derived probes were randomly hybridized to HumanRef-8 Expression v2 bead chip arrays (Illumina), followed by scanning and feature extraction (ServiceXS, Leiden, the Netherlands). Sample number and barcodes of the three monocyte-derived cell populations: (1) Unstimulated monocytes: N=9 nonresponding, N=10 responding patients, with barcodes GSM563265 to GSM563286. (2) 3h LPS monocytes: N=10 nonresponders, N=10 responders, with barcodes GSM335353 to GSM335389. (3) Macrophages: N=9 nonresponders, N=9 responders, with barcodes GSM563287 to GSM563304. The raw data for Samples GSM335xxx are provided as individual supplementary files. The raw data for Samples GSM563xxx are provided in the supplementary file "GSE13290_raw_data.txt".