Longrange PCR-based next-generation sequencing in pharmacokinetics and pharmacodynamics study of propofol among patients under general anaesthesia

Abstract

The individual response of patients to propofol results from the influence of genetic factors. However, the state of knowledge in this matter still remains insufficient. The aim of our study was to determine genetic predictors of variable pharmacokinetics and pharmacodynamics of propofol within selected 9 genes coding for propofol biotransformation enzymes, receptors and transporters. Our studies are the first extensive pharmaocgenetics research of propofol using high throughput sequencing technology. After the design and optimization of long range PCR-based next-generation sequencing experiment, we screened promoter and coding sequences of all genes analyzed among 87 Polish patients undergoing general anaesthesia with propofol. Initially we found that two variants, c.516 G > T in the CYP2B6 gene and c.2677 T > G in the ABCB1 gene, significantly correlate with propofol’s metabolic profile, however after Bonferroni correction the P-values were not statistically significant. Our results suggest, that variants within the CYP2B6 and ABCB1 genes correlate stronger with propofol’s metabolic profile compared to other 7 genes. CYP2B6 and ABCB1 variants can play a potentially important role in response to this anaesthetic and they are promising object for further studies.

NGS study of glucocorticoids response genes in inflammatory bowel disease patients

“…Therefore, as an additional goal of this work, we developed a rapid, flexible and economical application of NGS by combining it with LR-PCR as target enrichment for pharmacogenetic studies on GC drugs. A similar solution was used successfully in earlier studies [ 19 , 20 ]. In bioinformatic analysis for our scientific purposes, in contrast to diagnostic standards of minimum coverage 30× [ 26 ], we carefully applied a reads cover filter of 10 to avoid omission of any variants at this stage ( Figure 2 ).…”

“…Primers were designed as part of this work using the NCBI (National Center for Biotechnology Information) Primer-BLAST tool in reference to the human genomic sequence (GRCh37/hg19). The only exceptions were the primers and amplification conditions for the IL-1B, TNF and ABCB1 genes, which originated from our previous studies [19][20][21]. Due to the limitation of the LR-PCR method in the length of efficiently amplified DNA fragments, the LR-PCR of short genes such as IL-1A (11.48 kb), IL-1B (7.02 kb) and IL-2 (5.25 kb) was performed in one fragment, but the long genes, e.g.…”

Status Epilepticus During Recovery from General Anesthesia

“…Pharmacogenetic tests were conducted to understand whether there was a drug metabolism-related abnormality. Especially P-glycoprotein DNA variants might implicate a possible role in propofol metabolism [5,6]. CYP2C19 gene*2-*3-*17 haplotypes, CYP3A4 gene *1B haplotype and CYP2D6 gene *4 haplotype and P-glycoprotein (MDR1) gene c.3435C>T variant was investigated using peripheral blood DNA of the patient.…”

Precision Organ Function Protection for Enhanced Recovery after Surgery

“…Precision anesthesia requires experienced anesthesiologists to determine precise fluid therapy, suitable analgesia, and stationary hemodynamics according to changes and difficulties encountered during operation. To determine the sequence of drugs and the order of use, the anesthesiologist should be familiar with pharmacodynamics and pharmacokinetics of the drug, which may help their patients have a speedy recovery [84][85][86].…”

“…Traditional static stress monitoring indicators, such as CVP, PAWP, and ventricular end-diastolic volume indicators, cannot predict the effectiveness of fluid therapy. Therefore, it is necessary to find new hemodynamic indicators to monitor early fluid therapy [84][85][86].…”