{"id":1981,"date":"2020-05-05T17:34:24","date_gmt":"2020-05-05T15:34:24","guid":{"rendered":"https:\/\/www.newslab.sk\/identifikacia-metabolickych-ciest-v-patogeneze-diabetickej-retinopatie-s-vyuzitim-exomovych-sekvenacnych-dat-pilotna-studia\/"},"modified":"2020-05-05T17:37:43","modified_gmt":"2020-05-05T15:37:43","slug":"identification-of-metabolic-pathways-in-pathogenesis-of-diabetic-retinopathy-using-exome-sequencing-a-pilot-study","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/identification-of-metabolic-pathways-in-pathogenesis-of-diabetic-retinopathy-using-exome-sequencing-a-pilot-study\/","title":{"rendered":"Identification of metabolic pathways in pathogenesis of diabetic retinopathy using exome sequencing \u2013 a pilot study"},"content":{"rendered":"

*<\/strong>All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper.<\/strong><\/span><\/p>\n

 <\/p>\n

\u00davod<\/strong><\/p>\n

Diabetick\u00e1 retinopatia (DR) je jednou z naj\u010dastej\u0161\u00edch mikrovaskul\u00e1rnych komplik\u00e1ci\u00ed DM a rad\u00ed sa medzi naj\u010dastej\u0161ie pr\u00ed\u010diny straty zraku u \u013eud\u00ed stredn\u00e9ho a vy\u0161\u0161ieho veku. Vznik a progresia diabetickej retinopatie s\u00fa spojen\u00e9 s pred\u013a\u017een\u00fdm trvan\u00edm diabetu, hyperglyk\u00e9miou, hypertenziou a so zv\u00fd\u0161enou hladinou lipidov. Tieto faktory v\u0161ak predstavuj\u00fa len 10<\/p>\n

% rizika vzniku a rozvoja tohto ochorenia a predpoklad\u00e1 sa existencia \u010fal\u0161\u00edch faktorov, spomedzi ktor\u00fdch sa najv\u00e4\u010d\u0161\u00ed v\u00fdznam pripisuje genetick\u00fdm faktorom. Presn\u00fd mechanizmus vzniku ochorenia nie je jasn\u00fd, ale je zn\u00e1me, \u017ee do patogen\u00e9zy diabetickej retinopatie je zapojen\u00fdch viacero biochemick\u00fdch dr\u00e1h. Spom\u00ednan\u00e9 biochemick\u00e9 mechanizmy zah\u0155\u0148aj\u00fa napr\u00edklad polyolov\u00fa metabolick\u00fa dr\u00e1hu, neenzymatick\u00fa glyk\u00e1ciu a tvorbu koncov\u00fdch produktov pokro\u010dilej glyk\u00e1cie, oxida\u010dn\u00fd stres, aktiv\u00e1ciu prote\u00ednkin\u00e1zy C a mnoh\u00e9 in\u00e9(1).<\/p>\n

\u00da\u010das\u0165 genetick\u00fdch faktorov na vzniku DR je \u0161tudovan\u00e1 vo ve\u013ekej miere najm\u00e4 anal\u00fdzou variantov vybran\u00fdch kandid\u00e1tnych g\u00e9nov a v poslednom obdob\u00ed \u010doraz \u010dastej\u0161ie celogen\u00f3mov\u00fdmi asocia\u010dn\u00fdmi anal\u00fdzami (GWAS), ktor\u00e9 sa stali \u00fastrednou met\u00f3dou na \u0161t\u00fadium genetickej zlo\u017eky<\/p>\n

komplexn\u00fdch ochoren\u00ed, pri\u010dom umo\u017enili identifik\u00e1ciu mnoh\u00fdch asociovan\u00fdch chromoz\u00f3mov\u00fdch oblast\u00ed(2). Pri tak\u00fdchto \u0161t\u00fadi\u00e1ch je v\u00fdrazne limitovan\u00e9 mapovanie kauz\u00e1lnych variantov s malou popula\u010dnou frekvenciou a mal\u00fdm \u00fa\u010dinkom, ktor\u00e9 mo\u017eno identifikova\u0165 iba pri anal\u00fdzach ve\u013ek\u00fdch skup\u00edn, \u010dasto dosahuj\u00facich nieko\u013eko tis\u00edc testovan\u00fdch jedincov, \u010do v\u00fdrazne zvy\u0161uje laborat\u00f3rne a finan\u010dn\u00e9 n\u00e1roky tak\u00fdchto \u0161t\u00fadi\u00ed. \u010eal\u0161ou nev\u00fdhodou v konven\u010dnom pr\u00edstupe GWAS anal\u00fdz je vyu\u017eitie nek\u00f3duj\u00facich, \u010dasto medzig\u00e9nov\u00fdch jednonukleotidov\u00fdch (SNP) polymorfizmov, pomocou ktor\u00fdch v\u00e4zba k samotn\u00fdm kauz\u00e1lnym variantom nemus\u00ed by\u0165 v\u017edy spo\u013eahlivo odhalen\u00e1. V posledn\u00fdch rokoch sa preto v asocia\u010dn\u00fdch \u0161t\u00fadi\u00e1ch vyu\u017e\u00edvaj\u00fa pr\u00edstupy sekvenovania novej gener\u00e1cie, \u010dasto so zameran\u00edm na ex\u00f3m s mo\u017enos\u0165ou priamej identifik\u00e1cie kauz\u00e1lneho variantu(3). Napriek zapojeniu t\u00fdchto pokro\u010dil\u00fdch highthroughput postupov anal\u00fdzy DNA zost\u00e1va spojitos\u0165 genetick\u00fdch faktorov s diabetickou retinopatiou st\u00e1le ot\u00e1zna, preto\u017ee napriek mnoh\u00fdm \u0161t\u00fadi\u00e1m v\u00fdsledky nie s\u00fa konzistentn\u00e9, \u010do je sp\u00f4soben\u00e9 pravdepodobne najm\u00e4 v d\u00f4sledku multifaktori\u00e1lnej povahy ochorenia, tie\u017e v\u0161ak mal\u00e9ho po\u010dtu vzoriek v jednotliv\u00fdch \u0161t\u00fadi\u00e1ch a \u0165a\u017ekost\u00ed pri kontrole kovari\u00e1tov tejto choroby(3). Identifik\u00e1cia kauz\u00e1lnych variantov je v pr\u00edpade v\u00e4\u010d\u0161iny multifaktori\u00e1lnych ochoren\u00ed navy\u0161e komplikovan\u00e1 najm\u00e4 z d\u00f4vodu existencie a \u00fa\u010dasti variantov tzv. mal\u00e9ho \u00fa\u010dinku, ktor\u00fdch jednotliv\u00e1 identifik\u00e1cia je \u0165a\u017ek\u00e1 a na ich odhalenie je \u010dasto potrebn\u00e1 anal\u00fdza ich vz\u00e1jomn\u00fdch vz\u0165ahov. Z tohto d\u00f4vodu je jedn\u00fdm z pr\u00edstupov na objasnenie molekul\u00e1rnej patogen\u00e9zy vyu\u017eitie s\u00fa\u010dasnej anal\u00fdzy variantov vzh\u013eadom na ich pr\u00edslu\u0161nos\u0165 k rovnak\u00fdm metabolick\u00fdm cest\u00e1m. Anal\u00fdza metabolick\u00fdch ciest kombinuje efekt viacer\u00fdch variantov mal\u00e9ho \u00fa\u010dinku, a t\u00fdm zvy\u0161uje silu \u0161t\u00fadie(4). Tak\u00fdto pr\u00edstup m\u00f4\u017ee upriami\u0165 anal\u00fdzu na metabolick\u00fa cestu, v ktorej s\u00fa n\u00e1sledne sledovan\u00e9 aj in\u00e9 zriedkav\u00e9 varianty, ktor\u00e9 nemuseli by\u0165 nutne zachyten\u00e9 v prim\u00e1rnej asocia\u010dnej \u0161t\u00fadii. Z\u00e1rove\u0148 tento typ anal\u00fdzy odha\u013euje mo\u017en\u00fa spojitos\u0165 skupiny variantov s fenotypom, a teda pochopenie molekul\u00e1rnych mechanizmov, ktor\u00e9 je nevyhnutn\u00e9 na identifik\u00e1ciu biomarkerov na diagnostiku a lie\u010dbu.<\/p>\n

V tejto pilotnej \u0161t\u00fadii sme sa na z\u00e1klade anal\u00fdzy variantov z\u00edskan\u00fdch ex\u00f3mov\u00fdm sekvenovan\u00edm pacientov s diabetickou retinopatiou a kontrol spo\u010d\u00edvaj\u00facich z pacientov s DM bez diabetickej retinopatie zamerali na identifik\u00e1ciu mo\u017en\u00fdch kandid\u00e1tnych metabolick\u00fdch ciest pre \u010fal\u0161ie \u0161t\u00fadium tohto komplexn\u00e9ho ochorenia.<\/p>\n

 <\/p>\n

Experiment\u00e1lna \u010das\u0165<\/h1>\n

\u00a0<\/strong><\/p>\n

Materi\u00e1l a metodika<\/h2>\n

V\u0161etky genomick\u00e9 anal\u00fdzy boli uskuto\u010dnen\u00e9 na DNA vzork\u00e1ch perif\u00e9rnej krvi pacientov izolovan\u00fdch \u0161tandardn\u00fdmi automatizovan\u00fdmi postupmi (MagNA Pure LC DNA Isolation Kit, MagNA Pure LC 2.0 Instrument, Roche, \u0160vaj\u010diarsko) z\u00edskan\u00fdmi v r\u00e1mci rie\u0161enia projektu.<\/p>\n

Pr\u00edprava kni\u017en\u00edc pre ex\u00f3mov\u00e9 sekvenovanie bola uskuto\u010dnen\u00e1 pomocou Nextera Rapid Capture Exome kitu (Illumina) s ve\u013ekos\u0165ou cie\u013eovej sekvencie 37 Mb pod\u013ea odpor\u00fa\u010dan\u00ed v\u00fdrobcu. Jednotliv\u00e9 kroky pr\u00edpravy aj kvalita pripravenej kni\u017enice boli overen\u00e9 vyu\u017eit\u00edm kapil\u00e1rnej elektrofor\u00e9zy syst\u00e9mom Agilent 2100 Bioanalyzer s pou\u017eit\u00edm Agilent High Sensitivi- ty DNA Kitu (Agilent, USA). Pripraven\u00e9 kni\u017enice boli analyzovan\u00e9 syst\u00e9mom NextSeq 500 Illumina s vyu\u017eit\u00edm NextSeq 500 High Output Kitu. Analyzovan\u00e9 d\u00e1ta boli bioinformaticky spracovan\u00e9 pomocou web-based BaseSpace aplik\u00e1cie (Illumina) a GALAXY servera(5). Anal\u00fdza variantov aj identifik\u00e1cia variantov vykazuj\u00facich signifikantne rozdielne alelov\u00e9 zast\u00fapenie medzi skupinami pacientov boli uskuto\u010dnen\u00e9 pomocou Ingenuity variant analysis softv\u00e9ru (Qiagen) vyu\u017eit\u00edm asocia\u010dn\u00e9ho testu s bonferroniho korekciou. Anal\u00fdza zast\u00fapenia variantov v metabolick\u00fdch cest\u00e1ch bola vyhodnoten\u00e1 pomocou datab\u00e1zy ConsensusPathDB(6).<\/p>\n

 <\/p>\n

V\u00fdsledky<\/h2>\n

Ex\u00f3movou anal\u00fdzou bolo sk\u00faman\u00fdch 24 nepr\u00edbuzn\u00fdch pacientov s diabetom mellitom 2. typu rozdelen\u00fdch do dvoch skup\u00edn pod\u013ea pr\u00edtomnosti diabetickej retinopatie. 12 pacientov vo veku 40 a\u017e 65 rokov (priemer 52,5 roka) trpelo diabetom krat\u0161ie ako 6 rokov, pri\u010dom u nich boli diagnostikovan\u00e9 pr\u00edznaky neproliferat\u00edvnej formy diabetickej retinopatie s pr\u00edtomnos\u0165ou mikroaneuriziem, hemor\u00e1gi\u00ed a m\u00e4kk\u00fdch exud\u00e1tov. Zvy\u0161n\u00fdch 12 pacientov vo veku 48 a\u017e 84 rokov (priemer 65 rokov) predstavovali pacienti s diabetom<\/p>\n

 <\/p>\n

mellitom pretrv\u00e1vaj\u00facim viac ako 17 rokov, av\u0161ak bez n\u00e1lezu diabetickej retinopatie. V r\u00e1mci uskuto\u010dnenej ex\u00f3movej anal\u00fdzy s priemernou h\u013abkou \u010d\u00edtania cie\u013eov\u00fdch sekvenci\u00ed 150x bolo u pacientov identifikovan\u00fdch celkovo viac ako 120 000 variantov s minim\u00e1lnym pokryt\u00edm 20 \u010d\u00edtan\u00ed, z ktor\u00fdch 2 034 variantov, lokalizovan\u00fdch v 1 417 g\u00e9noch, vykazovalo signifikantne rozdielne alelov\u00e9 zast\u00fapenie medzi dvomi skupinami pacientov (s p-hodnotou \u2264 0,05, obr\u00e1zok 1). Samotn\u00e1 anal\u00fdza pr\u00edslu\u0161nosti identifikovan\u00fdch g\u00e9nov k metabolick\u00fdm cest\u00e1m (over-representation analysis) pomocou integrovanej datab\u00e1zy ConsensusPathDB odhalila spolu 166 metabolick\u00fdch ciest, ktor\u00e9 boli zast\u00fapen\u00e9 signifikantn\u00fdm po\u010dtom identifikovan\u00fdch g\u00e9nov. Identifikovan\u00e9 metabolick\u00e9 cesty poch\u00e1dzali z 11 datab\u00e1z. Medzi metabolick\u00e9 dr\u00e1hy, v ktor\u00fdch sa v signifikantne zv\u00fd\u0161enej miere vyskytovali g\u00e9ny obsahuj\u00face nami identifikovan\u00e9 varianty, patrili:<\/p>\n

O viazan\u00e1 glykozyl\u00e1cia <\/strong>(p = 2,05e-05), O-glykozyl\u00e1cia muc\u00ednov (p = 9,84e-05) a TSR prote\u00ednov (p = 0,0411) aj termin\u00e1cia biosynt\u00e9zy O-glyk\u00e1nov (p = 2,57e-05)<\/p>\n

Bunkov\u00e1 organiz\u00e1cia. <\/strong>Viacer\u00e9 metabolick\u00e9 cesty zodpovedn\u00e9 za organiz\u00e1ciu extracelul\u00e1rneho matrixu a interakciu re- ceptorov ECM (p = 2,31e-05, 0,00027), interakcie integr\u00ednov a beta-1 integr\u00ednu bunkov\u00fdch povrchov (p = 2,37e-05, 8,28e-05) \u010di interakcie lamin\u00ednov (p = 0,0154)<\/p>\n

Metabolizmus kolag\u00e9nu<\/strong>, jeho biosynt\u00e9zy (p = 2,86e-05) a for- m\u00e1cie (p = 9,47e-05) aj trimeriz\u00e1cie re\u0165azcov a degrad\u00e1cie (p = 9,96e-05, 0,00166)<\/p>\n

Metabolizmus lipidov, <\/strong>zast\u00fapen\u00fd viacer\u00fdmi cestami ako mitochondri\u00e1lna beta-oxid\u00e1cia dlh\u00fdch a stredne dlh\u00fdch re\u0165azcov (p = 0,00287, 0,00107), pred\u013a\u017eenie re\u0165azca mastn\u00fdch kysel\u00edn v mitochondrii (p = 0,00821) \u010di synt\u00e9za eikozanoidov (p = 0,00311) a in\u00e9.<\/p>\n

Metabolizmus plazmov\u00fdch lipoprote\u00ednov <\/strong>a ich kl\u00edrensu (p = 0,0123, 0,00525)<\/p>\n

Sign\u00e1lne dr\u00e1hy <\/strong>ako dr\u00e1ha ovplyv\u0148uj\u00faca hladinu cAMP cez G alfa podjednotky (p = 0,00639) \u010di NOTCH1 (p = 0,0143) a NOTCH3 (p = 0,0218) sign\u00e1lne dr\u00e1hy<\/p>\n

Metabolick\u00e9 cesty sp\u00e1jan\u00e9 so sel\u00e9nom <\/strong>a kyselinou sel\u00e9no- vou (p = 0,00109) a metabolizmus zl\u00fa\u010den\u00edn obsahuj\u00facich sel\u00e9n (p = 0,0225)<\/p>\n

Spomedzi 2 034 variantov vykazuj\u00facich signifikantne rozdielne zast\u00fapenie medzi skupinami boli identifikovan\u00e9 aj varianty v 32 g\u00e9noch, ktor\u00e9 zohr\u00e1vaj\u00fa d\u00f4le\u017eit\u00fa \u00falohu v patogen\u00e9ze retin\u00e1lnych ochoren\u00ed, napr\u00edklad CNGA1 (retinitis pigmentosa), ERCC6 (macular degeneration), NBAS (optic atrophy and retinal dystrophy), OPN1SW (tritanopia), POC5 (syndromic disease with retinitis pigmentosa), RP1L1, CDH3 (macular dystrophy) a in\u00e9.<\/p>\n

 <\/p>\n

Diskusia<\/h1>\n

Presn\u00fd mechanizmus vzniku diabetickej retinopatie doposia\u013e nie je napriek mnoh\u00fdm \u0161t\u00fadi\u00e1m celkom objasnen\u00fd,\u00a0 je v\u0161ak zn\u00e1me, \u017ee do patogen\u00e9zy je zapojen\u00fdch viacero biochemick\u00fdch dr\u00e1h pre regul\u00e1ciu hyperglyk\u00e9mie, hypertenzie, z\u00e1palu, hladiny lipidov a mnoh\u00e9 \u010fal\u0161ie. V tejto pilotnej \u0161t\u00fadii sme anal\u00fdzou d\u00e1t ex\u00f3mov\u00e9ho sekvenovania 24 pacientov identifikovali celkovo 166 kandid\u00e1tnych metabolick\u00fdch ciest, z ktor\u00fdch najviac zast\u00fapen\u00fdmi boli cesty zapojen\u00e9 do O-glykozyl\u00e1cie, organiz\u00e1cie ECM, metabolizmu kolag\u00e9nu, sel\u00e9nu, lipidov a plazmov\u00fdch lipoprote\u00ednov. Za jeden z hlavn\u00fdch faktorov vzniku DR je pova\u017eovan\u00fd pretrv\u00e1vaj\u00faci, subklinick\u00fd z\u00e1pal, ktor\u00fd v d\u00f4sledku hyperglyk\u00e9mie, oxida\u010dn\u00e9ho stresu, nadmernej glyk\u00e1cie a hypertenzie vedie k zv\u00fd\u0161en\u00e9mu tlaku retin\u00e1lnej vaskulat\u00fary, neovaskulariz\u00e1cii, hemor\u00e1gii a leukost\u00e1ze, ktor\u00e1 je v\u00fdznamn\u00fdm javom DR(7-10). Centr\u00e1lnu \u00falohu z\u00e1palu v patogen\u00e9ze DR potvrdzuj\u00fa aj \u0161t\u00fadie zameran\u00e9 na pod\u00e1vanie protiz\u00e1palov\u00fdch liekov, ktor\u00e9 br\u00e1nia progresii DR(11). Pretrv\u00e1vaj\u00faca expresia proinflama\u010dn\u00fdch cytok\u00ednov, chemok\u00ednov a aktiv\u00e1cia solubiln\u00fdch a bunkov\u00fdch adh\u00e9znych molek\u00fal koreluje s progresiou DR a prispieva k leukost\u00e1ze zvy\u0161ovan\u00edm interakcie leukocytov s endoteli\u00e1lnymi bunkami. K zv\u00fd\u0161enej adh\u00e9zii leukocytov prispievaj\u00fa zmeny v uhlovod\u00edkov\u00fdch re\u0165azcoch membr\u00e1n leukocytov, pri ktor\u00fdch v d\u00f4sledku z\u00e1palovej aktiv\u00e1cie N-acetylgluk\u00f3zaminyltransfer\u00e1zy (C2GNT) doch\u00e1dza k ich O-glykozyl\u00e1cii, ktorej zv\u00fd\u0161en\u00e1 miera koreluje s progresiou a z\u00e1va\u017enos\u0165ou DR aj diabetickej nefropatie(8). Lok\u00e1lny z\u00e1pal sp\u00f4sobuje tie\u017e aktiv\u00e1ciu mikrogli\u00e1lnych buniek, ktor\u00e1 vedie k zv\u00fd\u0161enej produkcii metaloprotein\u00e1z (MMPs). \u00dalohu MMPs vo vz\u0165ahu k organiz\u00e1cii extracelul\u00e1rneho matrixu a interakcii ECM receptorov analyzovali viacer\u00e9 \u0161t\u00fadie, pri ktor\u00fdch bolo preuk\u00e1zan\u00e9, \u017ee degrad\u00e1ciou komponentov ECM sa MMPs podie\u013eaj\u00fa na regul\u00e1cii prolifer\u00e1cie, neovaskulogen\u00e9zy a tkanivov\u00e9ho remodelingu(12,13). Varianty MMP-2 boli sledovan\u00e9 aj vo vz\u0165ahu ku kolag\u00e9nu IV, ktor\u00e9ho akumul\u00e1cia sp\u00f4sobuj\u00faca zv\u00e4\u010d\u0161enie baz\u00e1lnej membr\u00e1ny kapil\u00e1r je zn\u00e1mym patologick\u00fdm n\u00e1lezom DR a diabetickej nefropatie(14,15). \u00daloha sel\u00e9nu bola sk\u00faman\u00e1 v s\u00favislos- ti s regul\u00e1ciou enz\u00fdmov z\u00fa\u010dast\u0148uj\u00facich sa na ochrane proti oxida\u010dn\u00e9mu stresu, pri\u010dom bolo uk\u00e1zan\u00e9, \u017ee suplement\u00e1cia sel\u00e9nom pom\u00e1ha zmierneniu tvorby mikrovaskul\u00e1rnych l\u00e9zi\u00ed v retine a m\u00e1 protekt\u00edvny efekt pred oxida\u010dn\u00fdm stresom indukovan\u00fdm gluk\u00f3zou(16,17). Viacer\u00e9 hypot\u00e9zy vznikali oh\u013eadne \u00falohy plazmov\u00fdch lipoprote\u00ednov. K\u00fdm pri zdravej retine lipoprote\u00edny neprech\u00e1dzaj\u00fa krvno-retin\u00e1lnou bari\u00e9rou, v pr\u00edpade diabetickej retiny m\u00f4\u017ee by\u0165 t\u00e1to bari\u00e9ra naru\u0161en\u00e1, preto lipoprote\u00edny m\u00f4\u017eu hra\u0165 nepriamu, ale d\u00f4le\u017eit\u00fa \u00falohu v modul\u00e1cii n\u00e1sledkov tohto poru\u0161enia(18). S\u00favislos\u0165 DR a plazmov\u00fdch lipidov bola preuk\u00e1zan\u00e1 vo viacer\u00fdch \u0161t\u00fadi\u00e1ch priamym meran\u00edm koncentr\u00e1cie cholesterolu, LDL, HDL, LDL a HDL fosfolipidov, triglyceridov, a apolipoprote\u00ednov A a B, pri\u010dom bola pozorovan\u00e1 zv\u00fd\u0161en\u00e1 koncentr\u00e1cia viacer\u00fdch z nich pr\u00e1ve u pacientov s DR (19,20). V r\u00e1mci tejto pilotnej \u0161t\u00fadie boli identifikovan\u00e9 viacer\u00e9 metabolick\u00e9 cesty, ktor\u00e9 m\u00f4\u017eu by\u0165 kandid\u00e1tnymi pre \u010fal\u0161ie \u0161t\u00fadie, \u00faloha jednotliv\u00fdch g\u00e9nov a ich variantov v\u0161ak mus\u00ed by\u0165 sledovan\u00e1 anal\u00fdzou v\u00e4\u010d\u0161ieho s\u00faboru pacientov.<\/p>\n

 <\/p>\n

Po\u010fakovanie: <\/em><\/strong>Tento<\/em> \u010dl\u00e1nok vznikol v\u010faka podpore v r\u00e1mci OP V\u00fdskum a v\u00fdvoj pre projekt: Centrum v\u00fdskumu z\u00e1va\u017en\u00fdch ochoren\u00ed a ich komplik\u00e1ci\u00ed, ITMS 26240120038, spolufinancovan\u00fd zo zdrojov Eur\u00f3pskeho fondu region\u00e1lneho rozvoja.<\/em><\/p>\n

 <\/p>\n

 <\/p>\n

LITERAT\u00daRA<\/strong><\/p>\n

    \n
  1. Heng LZ, Comyn O, Peto T, et al. Diabetic retinopathy: Pathogenesis, clinical grading, management and future developments. Diabetic Medi- cine 2013; 30(6): 640-650.<\/li>\n
  2. Sharma A, Valle ML, Beveridge C, et al. Unraveling the role of genetics in the pathogenesis of diabetic retinopathy. Eye (Lond) 2019; 33(4): 534-541.<\/li>\n
  3. Han J, Lando L, Skowronska-Krawczyk D, et al. Genetics of Diabetic Retinopathy. Curr Diab Rep 2019; 19(9):<\/li>\n
  4. Kao PYP, Leung KH, Chan LWC, et Pathway analysis of complex diseases for GWAS, extending to consider rare variants, multi-omics and interactions. Biochim Biophys Acta Gen Subj 2017; 1861(2): 335-353<\/li>\n
  5. Afgan E, Baker D, Batut B, et The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2018 update. Nucle- ic Acids Res 2018; 46(W1): W537-W544.<\/li>\n
  6. Kamburov A, Stelzl U, Lehrach H, et The ConsensusPathDB interaction database: 2013 update. Nucleic Acids Res 2013; 41(Database is- sue): D793-800.<\/li>\n
  7. Chibber R, Ben-Mahmud BM, Chibber S, et al. Leukocytes in diabetic retinopathy. Current Diabetes Reviews 2007; 3(1): 3-14.<\/li>\n
  8. Chibber R, Ben-Mahmud BM, Coppini D, et al. Activity of the glycosylating enzyme, core 2 GlcNAc (beta1,6) transferase, is higher in polymorphonuclear leukocytes from diabetic patients compared with age- matched control subjects: relevance to capillary occlusion in diabetic retinopathy. Diabetes 2000; 49(10): 1724-1730.<\/li>\n
  9. Kowluru RA, Chan PS. Oxidative stress and diabetic retinopathy. Exp Diabetes Res 2007; 2007: 43603<\/li>\n
  10. Matthews DR, Stratton IM, Aldington SJ, et Risks of progression of retinopathy and vision loss related to tight blood pressure control in type 2 diabetes mellitus: UKPDS 69. DArch Ophthalmol 2004; 122(11): 1631-40.<\/li>\n
  11. Kern TS, Miller CM, Du Y, et Topical administration of nepafenac inhibits diabetes-induced retinal microvascular disease and underlying abnormalities of retinal metabolism and physiology. Diabetes 2007; 56(2): 373-379.<\/li>\n
  12. Mohammad G, Kowluru Novel role of mitochondrial matrix metalloproteinase-2 in the development of diabetic retinopathy. Invest Ophthalmol Vis Sci 2011; 52(6): 3832-3841.<\/li>\n
  13. Yang J, Fan XH, Guan YQ, et MMP-2 gene polymorphisms in type 2 diabetes mellitus diabetic retinopathy. Int J Ophthalmol 2010; 3: 137-140.<\/li>\n
  14. Alcolado JC, Baroni MG, Li SR, Galton Genetic variation around the collagen IV 1a gene locus and proliferative retinopathy in type 2 diabetes mellitus. Hum Hered 1993; 43(2): 126-130.<\/li>\n
  15. Arkkila PE, R\u00f6nnemaa T, Koskinen PJ, et Biochemical markers of type III and I collagen: association with retinopathy and neuropathy in type 1 diabetic subjects. Diabet Med 2001; 18(10): 816-821.<\/li>\n
  16. Gonz\u00e1lez de Vega R, Garc\u00eda M, Fern\u00e1ndez-S\u00e1nchez ML, et Protective effect of selenium supplementation following oxidative stress mediated by glucose on retinal pigment epithelium. Metallomics 2018; 10(1): 83-92.<\/li>\n
  17. Kowluru RA, Tang J, Kern Abnormalities of retinal metabolism in diabetes and experimental galactosemia. VII. Effect of long-term administration of antioxidants on the development of retinopathy. Diabetes 2001; 50(8): 1938-1942.<\/li>\n
  18. Yu JY, Lyons Modified Lipoproteins in Diabetic Retinopathy: A Local Action in the Retina. J Clin Exp Ophthalmol 2013; 4(6): pii: 314.<\/li>\n
  19. Sinav S, Onelge MA, Onelge S,et Plasma lipids and lipoproteins in retinopathy of type I (insulin-dependent) diabetic patients. Ann Ophthalmol 1993; 25(2): 64-66.<\/li>\n
  20. Malaguarnera G, Gagliano C, Bucolo C, et al. Lipoprotein(a) serum levels in diabetic patients with retinopathy. Biomed Res Int 2013; 2013: 943505.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    *All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper.   \u00davod Diabetick\u00e1 retinopatia (DR) je jednou z naj\u010dastej\u0161\u00edch mikrovaskul\u00e1rnych komplik\u00e1ci\u00ed DM a rad\u00ed sa medzi naj\u010dastej\u0161ie pr\u00ed\u010diny straty zraku u \u013eud\u00ed stredn\u00e9ho a vy\u0161\u0161ieho veku. Vznik a progresia diabetickej<\/p>\n","protected":false},"author":7,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_mi_skip_tracking":false,"footnotes":""},"categories":[290],"tags":[1456,735,1457,1459,1458],"class_list":["post-1981","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-analyza-metabolickych-ciest-en","tag-diabetic-retinopathy","tag-diabeticka-retinopatia-en","tag-exome-sequencing","tag-exomove-sekvenovanie-en","typ_clanku-original-work"],"acf":{"abstrakt":"

    Diabetic retinopathy is one of the most common microvascular complications of diabetes mellitus and is a com- mon cause of vision loss in western countries. Hyperglycaemia, hypertension and dyslipidaemia, which are among the main factors, however, represent only approximately 10% of total risk of development and disease progression. Various genetic variants are considered and tested intensively to unveil the exact pathophysiological mechanism. In this pilot study, we analysed the representation of DNA variants identified by exome sequencing in the group of patients with diabetic retinopathy and diabetic patients without retinopathy, to identify metabolic pathways that could play a role in the disease onset. Among the significant pathways those involved in O-gly- cosylation, metabolic pathways of cellular organization and components, metabolism of collagen, lipids, plasma lipoprotein, and selenium were most represented. Metabolic pathway analysis combines the weak effects of multiple variants and increases the power of information mainly in association studies conducted on a small number of samples.<\/p>\n

    Keywords: <\/strong>diabetic retinopathy, exome sequencing<\/p>\n","casopis":[{"ID":1893,"post_author":"7","post_date":"2020-05-05 11:32:54","post_date_gmt":"2020-05-05 09:32:54","post_content":"

      \r\n \t
    • Identification of metabolic pathways in pathogenesis of diabetic retinopathy using exome sequencing \u2013 a pilot study<\/li>\r\n \t
    • Anti-tumour effects of vitamin D<\/li>\r\n \t
    • Molecular detection methods of mutations in the kinase domain of fusion gene bcr-abl1 in patients with chronic myelocyte leukemia<\/li>\r\n \t
    • The case report of toxoplasmic meningoencephalitis with fatal outcome in HIV patient<\/li>\r\n \t
    • Carcinosarcoma-like endometrioid carcinoma of the uterus: case report of rare non-high-grade tumor<\/li>\r\n<\/ul>","post_title":"newsLab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-4","to_ping":"","pinged":"","post_modified":"2020-05-05 15:13:41","post_modified_gmt":"2020-05-05 13:13:41","post_content_filtered":"","post_parent":0,"guid":"https:\/\/www.newslab.sk\/?post_type=casopis&p=1893","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"5-8","upload_clanok":{"ID":1979,"id":1979,"title":"NEWSLAB_1-2020_Ficek","filename":"NEWSLAB_1-2020_Ficek.pdf","filesize":192840,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2020\/05\/NEWSLAB_1-2020_Ficek.pdf","link":"https:\/\/www.newslab.sk\/en\/identification-of-metabolic-pathways-in-pathogenesis-of-diabetic-retinopathy-using-exome-sequencing-a-pilot-study\/newslab_1-2020_ficek-2\/","alt":"","author":"7","description":"","caption":"","name":"newslab_1-2020_ficek-2","status":"inherit","uploaded_to":1981,"date":"2020-05-05 15:23:47","modified":"2020-05-05 15:23:47","menu_order":0,"mime_type":"application\/pdf","type":"application","subtype":"pdf","icon":"https:\/\/www.newslab.sk\/wp-includes\/images\/media\/document.png"}},"_links":{"self":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1981","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/users\/7"}],"replies":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/comments?post=1981"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1981\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1981"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1981"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1981"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}