PhD Scientific Days 2022

Translational Medicine II. (Poster discussion will take place in the Aula during the Coffee Break)

Novel Druggable Targets for Heart Failure Identified by Droplet Digital PCR and Next Generation Sequencing

Text of the abstract

Introduction: Despite the currently available potent therapies of heart faiulure (HF), identification of novel druggable targets is still needed to improve outcomes for patients’ benefit. G-protein coupled receptors (GPCRs) represent the largest family of targets for already approved drugs providing a great opportunity for drug repurposing. Therefore, screening for differences in cardiac GPCR expression during HF provides an efficient way of identifying novel drug targets for this disease. We hypothesized that droplet digital PCR (ddPCR) providing an absolute quantification of mRNAs in a highly sensitive manner can be used as a screening method to identify differentially expressed GPCRs in failing vs. healthy hearts.
Aim: Here we aimed to investigate the differential expressions of 288 cardiac GPCRs by ddPCR and Next Generation Sequencing (NGS) in a rat model of HF.
Methods: 8-10 weeks old, male Wistar rats were subjected to transverse aortic constriction (TAC, n=5) or sham (SHAM, n=5) surgery. 15-18 weeks after surgery, cardiac echocardiography was performed, and cardiac samples were collected for histological and gene expression analyses. Absolute quantification of the expression of 288 GPCR genes was performed by ddPCR, and by bulk RNA sequencing using NGS. Differential expression of GPCRs of TAC and SHAM hearts identified by both techniques was compared, and the results were correlated.
Results: TAC animals showed significantly deteriorated systolic and diastolic functions, accompanied by cardiac hypertrophy and fibrosis when compared to SHAM. Out of 288 GPCRs, ddPCR identified a total of 27 genes, and NGS identified a total of 69 genes to be significantly differently expressed in TAC vs. SHAM animals, 14 of which were identified by both methods, showing significant and clinically meaningful correlation.
Conclusions: This is the first demonstration of using ddPCR as a screening method for identifying GPCRs as novel druggable targets in a disease, and validated our results by NGS. As GPCRs are potent candidates for drug repurposing, we provide a list of easily testable drug targets for future basic research in the field of HF.
Funding: NVKP_16-1-2016-0017; 2020-4.1.1.-TKP2020; EFOP-3.6.3-VEKOP-16-2017-00009; Gedeon Richter Excellence PhD Scholarship; European Union’s Horizon 2020 Research and Innovation Programme under grant agreement no. 739593