The primary aim of this Work Package is to bundle the efforts of the work packages on the Russian side of the consortium, and, together with WP10 on the European side, to provide a platform for research that makes use of large blood sample banks into type 2 diabetes, cachexia, and obesity as co-morbidities in heart failure. Due to the large number of patients (estimated more than 1000 patients with heart failure) that will be enrolled into the project, the Blood and DNA Banks in MSU and AC will by use of innovative and collaborative approaches enable research to investigate and establish novel genetic markers or panels of genetic markers to better characterize heart failure patients with one or more of the aforementioned co-morbidities in terms of their prognosis, their clinical course (e.g., imminent decompensation of heart failure), and the verification of their clinical diagnosis.
Estimation of the patients' prognosis and their clinical course are important for clinical decision making, because patients with more advanced disease or at risk of imminent decompensation may benefit from more aggressive therapies or be candidates for novel therapies (i.e. cell therapy). Therefore, close collaboration with other WPs (especially WP13 and WP14) that collect detailed clinical information about the patients and their clinical course over time, is necessary. Since several studies over the last years have shown that different co-morbidities affect the reliability of "gold standard" biomarkers in heart failure, the search for independent marker(s) allowing to predict patient prognosis is necessary. Genetic markers (gene polymorphisms and mutations) can serve as reliable predictors for poor prognosis for heart failure patients with obesity, cachexia, and type 2 diabetes. Thus, mutant phenotype of MTHFR is associated with early hypertension in obese patients (Andreenko et al., 2007), mutations in eNOS gene could serve as a marker for poor prognosis for patients with atherosclerosis (Kuznetsova et. al., 2007). Furthermore, benefits of cell therapy for heart failure treatment may depend on genetic factors, which influence the functional activity of cells utilized for the therapy.
Biomaterial in the Blood and DNA Banks will enable research into the impact of type 2 diabetes, obesity, and cachexia on established biomarkers of heart failure prognosis, severity, and the clinical course of the disease. A large-scale collection of EDTA plasma and genomic DNA samples requires the setting up of a standardised -80░C storage system using existing freezers that allows safe and reliable storage of biomedical samples under standardised conditions until analysis. All initial samples will be stored in 700 microlitre aliquots of EDTA plasma in 1,5 ml safe-lock Eppendorf tubes (in triplicates). This procedure allows safe long-term storage. Labelling is supposed to make use of a real-text label on the sample front with the patient's number which corresponds to patient data, including surname, given name, date of birth and clinical details and study date. All procedures will be held within the tight borders of the ethics approval. DNA isolation will be done using robotic system (QiaCube, Qiagen), which allow to obtain good quality DNA samples and guarantees that sample identity remains uncompromised.
Task 1. To search for new heart-failure associated genetic markers
Several genetic markers have been shown in previous studies to be associated with metabolic syndrome, type 2 diabetes, obesity and fewer with cachexia. Mostly they are genes involved in lipid metabolism (ADIPOQ, RSTN, PPAR, APOA5, APOC3, FTO), as well as inflammatory cytokines (IL)-6, TNF-#) and hormones glucocorticoids, estrogenes). Their functional and prognostic impact has been not studied in association with heart failure development and prognosis. We will study the genetic polymorphisms, associated with metabolic syndrome, type 2 diabetes, and obesity in patients with the above mentioned co-morbidities of heart failure and, importantly, to correlate the obtained data on genetic polymorphisms with functional cellular, morphological and molecular biology studies performed within the frame of other work packages (collaborative networking particularly with WP12 and WP14. Use the DNA and biomarkers data from the Central Blood and DNA Bank of WP10).
Task 2. To test known genetic markers associated with ischemic heart disease and HF in relation to poor prognosis for patients with HF complicated by type 2 diabetes, obesity and cachexia
Several genetic markers have been shown in previous studies to be associated with risk and prognosis estimation of heart failure. Most of them represent functional or silent genetic polymorphisms among the genes involved in the adrenergic system (ADRB1, ADRB2, A2CAR), vascular physiology (MTHFR, prothrombin, factor V, GP IIb/IIIa, PAI1, factor VII, factor XIII) and endothelial metabolism (BDKRB2, EDN1, VEGF, eNOS), the renin-angiotensin-aldosterone system (ACE, AGT), oxidative stress (p22phox) and the inflammatory response (TNF#). However, their simultaneous effect on heart failure clinical cause and it's comorbidities has not been extensively studied. The above mentioned as well as other reported genetic polymorphisms will be studied for their association with heart failure prognosis, obesity and cachexia, as well as for their functional impact on the morphological, cellular and molecular level (collaborative networking with WP7, WP8, WP12 and WP14). Use the DNA and biomarkers data from biobank WP10).