Povezanost izvanstaničnih molekula proteina toplinskoga šoka 70 i adenozin-trifosfata sa sustavnim upalnim odgovorom u pacijenata s kroničnom opstrukcijskom plućnom bolesti

doctoral thesis

• Title (english): Association of extracellular molecules heat shock protein 70 and adenosine triphosphate with systemic inflammatory response in patients with chronic obstructive pulmonary disease
• Author: Iva Hlapčić
• Mentor: Lada Rumora (mentor)
• Committee member: Karmela Barišić (predsjednik povjerenstva)
• Committee member: Sanja Popović-Grle (član povjerenstva)
• Committee member: Dunja Rogić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2022, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry
• Universal decimal classification (UDC): 615 - Pharmacology. Therapeutics. Toxicology

Abstract

Kronična opstrukcijska plućna bolest (KOPB) kompleksna je i heterogena bolest s kroničnom upalom. Sustavni upalni odgovor vidljiv je u povećanju koncentracije općih upalnih biljega poput C-reaktivnog proteina (CRP), fibrinogena (Fbg), ukupnog broja leukocita (lkc) i upalnih citokina. Uslijed upalnih reakcija i oštećenja stanica u KOPB-u, povećava se koncentracija izvanstaničnog proteina toplinskoga šoka 70 (eHsp70) i izvanstaničnog adenozin-trifosfata (eATP). U ovom istraživanju određene su plazmatske koncentracije eHsp70 i eATP-a u 137 pacijenata sa stabilnim KOPB-om i uspoređene s koncentracijama u 95 kontrolnih ispitanika. Ispitano je postoji li povezanost ovih parametara sa stupnjem plućne opstrukcije, simptomima i povijesti egzacerbacija te pušačkim statusom. Određena je relativna razina ekspresije gena HSP70, TLR2 i TLR4 (dva receptora za eHsp70) te P2X7R i P2Y2R (dva receptora za eATP) te genotipizacija polimorfizama u genima HSP70, TLR2 i TLR4. eHsp70 i eATP povezani su s KOPB-om, sa stupnjom plućne opstrukcije i progresijom simptoma. Također, koncentracije eHsp70 i eATP-a veće su u skupini zdravih pušača nego u zdravih nepušača što ukazuje na pojavu upalnih procesa prilikom izlaganja cigaretnom dimu prije pojave bolesti. Značajna povezanost uočena je između IL-1β, IL-6, TNF-α, CRP-a, Fbg-a, eHsp70 i eATP-a u pacijenata s KOPB-om, a hijerarhijskom klasterskom analizom potvrđena je heterogenost i kompleksnost bolesti. Model sastavljen od IL-1β, eHsp70 i eATP-a pokazao je veliku predikcijsku vrijednost. Značajno povećanje relativne genske ekspresije u pacijenata s KOPB-om u odnosu na zdrave ispitanike opaženo je za HSP70, TLR2 i P2Y2R gene, dok je analiza polimorfizama ukazala da postoji povezanost između rizika od KOPB-a i polimorfizma rs6457452 u promotorskom dijelu gena HSP70. eHsp70 i eATP biljezi su sustavnog upalnog odgovora u KOPB-u i dio su patogeneze KOPB-a, a u kombinaciji s IL-1β čine model s velikim predikcijskim potencijalom. Dodatni doprinos istraživanja postignut je analizom genske ekspresije i polimorfizama koje pružaju bolje razumijevanje mehanizama i patogeneze KOPB-a.

Abstract (english)

Chronic obstructive pulmonary disease (COPD) is a complex and heterogenous disease with chronic inflammation. Systemic inflammatory response is followed by increased concentration of common inflammatory biomarkers as C-reactive protein (CRP), total count of leukocytes (lkc), fibrinogen (Fbg) and inflammatory cytokines. There is an increase in extracellular heat shock protein 70 (eHsp70) and extracellular adenosine triphosphate (eATP) due to inflammatory response and cell damage in COPD. In this thesis, eHsp70 and eATP were determined in 137 patients with stable COPD and compared with the concentrations in 95 controls. It was assessed if there is the association between eHsp70 and eATP and the severity of the airflow limitation, symptoms burden and history of excerbations as well as smoking status. The relative gene expression HSP70, TLR2 and TLR4 (eHsp70 receptors) as well as P2X7R and P2Y2R (eATP receptors) was determined. Moreover, selected HSP70, TLR2 i TLR4 polymorphisms were genotyped. There was an association of eHsp70 and eATP with COPD compared to healthy subjects, airflow limitation and symptoms severity. eHsp70 and eATP were increased in control smokers compared to non-smoking controls which indicates there is an inflammatory response due to the expose of cigarette smoke even before the disease occurs. Significant association was observed between IL-1β, IL-6, TNF-α, CRP, Fbg, eHsp70 and eATP in COPD patients, while hierarchical cluster analysis confirmed heterogeneity and complexity of COPD. Based on logistic regression analysis, combination of IL-1β, eHsp70 and eATP-a showed significant predictive potential in identifying COPD subjects. There was a significant increase in the relative gene expression of HSP70, TLR2 and P2Y2R genes in COPD patients, while the analysis of single nucleotide polymorphisms showed an association of rs6457452 in the promoter region of HSP70 with the risk of COPD. In conclusion, eHsp70 and eATP are biomarkers of systemic inflammatory response in COPD and it seems they might be useful as prognostic biomarkers, especially in a combination with IL-1β. This thesis contributed with the data at genetic level which could be useful for better understanding of the COPD pathogenesis.

• Keywords: kronična opstrukcijska plućna bolest; sustavna upala; izvanstanični protein toplinskoga šoka 70; izvanstanični adenozin-trifosfat; genska ekspresija; polimorfizam jednog nukleotida
• Keywords (english): chronic obstructive pulmonary disease; systemic inflammation; extracellular heat shock protein 70; extracellular adenosine triphosphate; gene expression; single nucleotide polymorphism
• Language: croatian
• URN:NBN: urn:nbn:hr:163:468319
• Promotion: 2022
• Study programme: Title: Pharmacy and biochemistry; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor znanosti (doktor znanosti)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-07-05 08:44:10