Naslov (srp)

Uticaj antiretrovirusnih lekova i genskih varijanti TERT i MTHFR gena na dužinu telomera kod pacijenata sa HIV infekcijom : doktorska disertacija

Autor

Bukić, Ena, 1985-

Doprinosi

Dragović-Lukić, Gordana, 1972-
Novaković, Ivana, 1961-
Gojković-Bukarica, Ljiljana, 1962-
Milašin, Jelena, 1957-
Vukmirović, Saša, 1979-
Ranin, Jovan, 1963-

Opis (srp)

Uvod: Uprkos efikasnoj kombinovanoj antiretrovirusnoj terapiji (combined antiretroviral therapy, cART), u organizmu pacijenata koji su inficirani humanim virusom imunodeficijencije (human immunodeficiency virus, HIV), formiraju se anatomski rezervoari u kojima se virus i dalje replikuje, te je na ovaj način uticaj HIV virusa na organizam hronično prisutan. Inflamacija i kumulativna toksičnost usled kontinuirane primene cART su potencijalno povezani sa ubrzanim procesom starenja. Biološka starost HIV-om inficiranih pacijenata može odstupati od njihove hronološke starosti. Biološka starost pacijenata se može pratiti promenama vrednosti biomarkera starenja. Pouzdanim, savremenim molekularnim biomarkerom starenja organizma smatra se promena dužine telomera. Telomere su DNK sekvence na krajevima hromozoma koje štite hromozome od gubitka genetičkog materijala. Tokom života, telomere se skraćuju prilikom svake ćelijske deobe. Kada se dostigne kritično skraćenje telomera, deobe ćelija se zaustavljaju i dolazi do programirane ćelijske smrti. Na dužinu telomera negativno mogu uticati primena cART i hronična inflamacija. Takođe, potencijalnu asocijaciju sa promenom dužine telomera pokazuju polimorfizmi rs2736100 i rs2736098 u TERT (telomerase reverse transcriptase) genu i polimorfizmi rs1801133 i rs1801131 u MTHFR (methylenetetrahydrofolate reductase) genu.Cilj: Ispitati povezanost dužine telomera kod HIV-om inficiranih pacijenata sa dužinom trajanja infekcije HIV-om, dužinom trajanja cART-a, mehanizmom delovanja primenjenih lekova u sklopu cART-a, genskim varijantama rs2736100 i rs2736098 TERT gena i genskim varijantama rs1801133 i rs1801131 MTHFR gena. Ispitati dijagnostički potencijal dužine telomera kao relevantnog parametra koji bi mogao uticati na odabir terapije HIV-om inficiranih pacijenata.Metode: U ovu studiju preseka je uključeno 176 pacijenata muškog pola, starosti iznad 18 godina, koji primaju cART u trajanju od minimum 16 nedelja i imaju manje od 50 kopija HIV RNK/ml krvi. Venepunkcijom se od pacijenata sa dijagnostikovanom HIV infekcijom, koji se leče u Centru za HIV/AIDS Klinike za infektivne i tropske bolesti u Beogradu, Univerzitetskog kliničkog centra Srbije, uzimalo po 4 ml uzorka krvi za farmakogenetsko ispitivanje. Po izvršenoj inaktivaciji virusa, iz uzoraka krvi, su izdvojene mononuklearne ćelije iz kojih je metodom isoljavanja izolovana DNK. RTL je određena metodom lančane reakcije polimeraze u realnom vremenu (real-time PCR), u odnosu na referentnu sekvencu. Utvrđivanje genotipova TERT i MTHFR polimorfizama je određeno metodom lančane reakcije polimeraze (PCR), praćene enzimskom digestijom PCR produkata (restriction fragment length polymorphisms-RFLP). Zdravstveni podaci relevantni za istraživanje su prikupljeni iz zdravstvenih kartona i istorije bolesti pacijenata. Klasifikacija individualnih parametara, dobijenih kliničkim i laboratorijskim ispitivanjima, njihova statistička obrada i grafički prikaz rezultata izvršeni su pomoću specijalizovanih R-softverskih paketa. Mašinskim učenjem pomoću softverskih paketa “randomForest” i “Boruta” izdvojene su varijable za inferencijalno statističko testiranje asocijacije sa RTL. Inferencijalna statistika je obuhvatila testove prema vrsti varijabli: Man-Vitnijev U test i Kruskal-Wallis test za obradu podataka u slučajevima odsustva normalne raspodele. Odnos kategorijalnih i neprekidno promenjivih varijabli testiran je Spirmanovim ili Kendalovim testom sa određivanjem koeficijenta korelacije. Za sva statistička testiranja, prag značajnosti je bio na konvencionalnom nivou od 0.05, a snaga statističkog testa iznosila je 0.8...

Opis (srp)

Medicina - Medicinska farmakologija / Medicine- Medical pharmacology Datum odbrane: 30.09.2024.

Opis (eng)

Introduction: Despite effective combined antiretroviral therapy (cART), in the body of patients infected with the human immunodeficiency virus (HIV), anatomical reservoirs are formed in which the virus continues to replicate, and in this way the influence of the HIV virus on the body is chronically present. Inflammation and cumulative toxicity from continuous cART administration are potentially associated with an accelerated aging process. The biological age of HIV-infected patients may differ from their chronological age. The biological age of patients can be monitored by changes in the values of aging biomarkers. Change in the length of telomeres is considered a reliable, modern molecular biomarker of the aging of the organism. Telomeres are DNA sequences at the ends of chromosomes that protect chromosomes from loss of genetic material. During life, telomeres shorten with each cell division. When a critical telomere shortening is reached, cell divisions stop and programmed cell death occurs. Telomere length can be negatively affected by cART administration and chronic inflammation. Also, polymorphisms rs2736100 and rs2736098 in the TERT (telomerase reverse transcriptase) gene and polymorphisms rs1801133 and rs1801131 in the MTHFR (methylenetetrahydrofolate reductase) gene show a potential association with the change in telomere length.Objective: To examine the association of telomere length in HIV-infected patients with the duration of HIV infection, the duration of cART, the mechanism of action of the drugs administered as part of cART, gene variants rs2736100 and rs2736098 of the TERT gene and gene variants rs1801133 and rs1801131 MTHFR gene. To examine the diagnostic potential of telomere length as a relevant parameter that could influence the choice of therapy for HIV-infected patients.Methods: This cross-sectional study included 176 male patients, aged over 18 years, receiving cART for a minimum of 16 weeks and having less than 50 copies of HIV RNA/ml blood. A 4 ml blood sample for pharmacogenetic testing was taken by venipuncture from patients diagnosed with HIV infection, who are being treated at the Center for HIV/AIDS, Clinic for Infectious and Tropical Diseases in Belgrade, University Clinical Center of Serbia. After the inactivation of the virus, mononuclear cells were isolated from the blood samples, from which DNA was isolated using the salting method. The RTL was determined by the polymerase chain reaction method in real time (real-time PCR), in relation to the reference sequence. Determining the genotypes of TERT and MTHFR polymorphisms was determined by the method of polymerase chain reaction (PCR), followed by enzymatic digestion of PCR products (restriction fragment length polymorphisms-RFLP). Health data relevant to the study were collected from patients' medical records and medical histories. The classification of individual parameters obtained from clinical and laboratory tests, their statistical processing and graphical presentation of the results were performed using specialized R-software packages. Using machine learning, i.e. "randomForest" and "Boruta" software packages, variables were extracted for inferential statistical testing of association with RTL. Inferential statistics included tests according to the type of variables: Mann-Whitney U test and Kruskal-Wallis test for data processing in cases of absence of normal distribution. The relationship between categorical and continuously changing variables was tested by Spearman's or Kendall's test with the determination of the correlation coefficient. For all statistical tests, the significance threshold was at the conventional level of 0.05, and the power of the statistical test was 0.8...

Jezik

srpski

Datum

2024

Licenca

Ovo delo je licencirano pod uslovima licence
Creative Commons CC BY-NC-ND 3.0 AT - Creative Commons Autorstvo - Nekomercijalno - Bez prerada 3.0 Austria License.

http://creativecommons.org/licenses/by-nc-nd/3.0/at/legalcode

Predmet

OSNO - Opšta sistematizacija naučnih oblasti, Farmakologija

HIV, aging, cART, telomeres, RTL

OSNO - Opšta sistematizacija naučnih oblasti, Farmakologija

HIV, starenje, cART, telomere, RTL

616.98:615.281.8(043.3)