Cardiovascular pharmacogenomics-related genetic biomarkers: electrochemical genosensors for personalized medicine

Barbosa, Tiago Manuel Moreira

Publication

Cardiovascular pharmacogenomics-related genetic biomarkers: electrochemical genosensors for personalized medicine

2024-03-25Master thesis

dc.contributor.advisor	Barroso, M. Fátima
dc.contributor.advisor	Santos, Marlene
dc.contributor.advisor	Vieira, Mónica
dc.contributor.author	Barbosa, Tiago Manuel Moreira
dc.date.accessioned	2025-02-17T14:17:32Z
dc.date.available	2025-02-17T14:17:32Z
dc.date.issued	2024-03-25
dc.date.submitted	2024-03-25
dc.description.abstract	Cardiovascular diseases (CVDs) are one of the leading causes of death worldwide, and the anticoagulant warfarin is routinely used to treat and control CVDs. However, due to several individual factors, including genetic variations, as singles nucleotide polymorphisms (SNPs), drug’s efficacy varies between individuals. Moreover, given warfarin’s narrow therapeutic index, genotype-guided approaches have become critical in establishing personalized doses. Since conventional technologies for SNP detection such as Polymerase Chain Reaction or Real-Time Polymerase Chain Reaction (PCR or RT-PCR) are costly, time-consuming and demand skilled specialists, novel disposable electrochemical genosensors have been developed and optimized to facilitate SNP detection. In this project, two electrochemical genosensors were designed to detect both the -1639 G>ASNP in the vitamin K epoxide reductase complex (VKORC1) gene, important in the anticoagulant effect, and the CYP2C9 g.9133C>T gene variation involved in the metabolization of warfarin. The construction of these genosensors employed the formation of binary mixed self-assembled monolayers (SAMs) composed by DNA capture probes and mercaptohexanol (MCH), immobilized onto gold screen-printed electrodes (AuSPE). Furthermore, these genosensors employed a sandwich format hybridization technique (to increase sensitivity) that included the use of a fluorescein-labeled signaling probe and the enzymatic amplification of the electrochemical signal, measured via chronoamperometry. To obtain genomic DNA useful to be used in the genosensors, PCR technique was used to amplify genomic DNA from biological samples. Then, these samples were tested in the genosensors. The genosensors were able to detect genomic DNA associated to the CYP2C9 and VKORC1 genes, but only distinguished SNPs existent in the CYP2C9 gene displaying the ability to help in establishing the optimal warfarin dosage, providing a cost-effective alternative to the conventional procedures. This technique has the potential to improve patient care in cardiovascular medicine by tackling the problems associated with CVD therapy and the genetic heterogeneity of medication response.	por
dc.description.sponsorship	CYTED—GENOPSYSEN, P223RT0141; COST, ENOTTA-CA21147
dc.identifier.tid	203852567
dc.identifier.uri	http://hdl.handle.net/10400.22/29538
dc.language.iso	eng
dc.relation	FCT/CAPES 2018/2019 Processo 4.4.1.00; CAPES (CAPES-FCT2017484310P 200.137.174.210); UIDB/50006/2020, UIDP/50006/2020, LA/P/0008/2020
dc.rights.uri	N/A
dc.subject	Cardiovascular diseases
dc.subject	Warfarin
dc.subject	Polymorphisms
dc.subject	VKORC1
dc.subject	CYP2C9
dc.subject	PCR
dc.subject	Electrochemical genosensors
dc.title	Cardiovascular pharmacogenomics-related genetic biomarkers: electrochemical genosensors for personalized medicine	por
dc.type	master thesis
dspace.entity.type	Publication