{"id":1582,"date":"2018-11-11T14:52:20","date_gmt":"2018-11-11T13:52:20","guid":{"rendered":"http:\/\/www.newslab.sk\/2018\/11\/11\/cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-prediktivnou-a-diagnostickou-hodnotou-u-pacientov-s-nadorovymi-ochoreniami\/"},"modified":"2018-11-11T14:54:26","modified_gmt":"2018-11-11T13:54:26","slug":"cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-prediktivnou-a-diagnostickou-hodnotou-u-pacientov-s-nadorovymi-ochoreniami","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-prediktivnou-a-diagnostickou-hodnotou-u-pacientov-s-nadorovymi-ochoreniami\/","title":{"rendered":"Circulating tumor DNA and its utilization as marker with prognostic, predictive and diagnostic value in patients with oncological diseases"},"content":{"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.<\/span><\/strong><\/p>\n

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Cirkuluj\u00faca n\u00e1dorov\u00e1 DNA<\/strong><\/p>\n

Zv\u00fd\u0161en\u00e1 hladina cirkuluj\u00facich nukleov\u00fdch kysel\u00edn u pacientov s rakovinou bola prv\u00fdkr\u00e1t op\u00edsan\u00e1 u\u017e v roku 1977(1). Trvalo \u010fal\u0161\u00edch 10 rokov, pok\u00fdm Stroun a kol. preuk\u00e1zali pr\u00edtomnos\u0165 neoplastick\u00fdch charakterist\u00edk v cirkul\u00e1cii, ke\u010f zistili, \u017ee cirkuluj\u00faca DNA vykazuje ur\u010dit\u00fa dvojvl\u00e1knov\u00fa instabilitu, ktor\u00e1 je \u0161pecifick\u00e1 pre n\u00e1dorov\u00fa DNA(2). Definit\u00edvny d\u00f4kaz, \u017ee vo\u013en\u00e1 cirkuluj\u00faca DNA v plazme onkologick\u00fdch pacientov je n\u00e1dorov\u00e9ho p\u00f4vodu, bol dosiahnut\u00fd po tom, ako Sorenson a kol. a Vasioutkin a kol. detegovali KRAS <\/em>mut\u00e1cie v cirkul\u00e1cii pacientov s rakovinou pankreasu(3), respekt\u00edve NRAS <\/em>mut\u00e1cie u pacientov s ak\u00fatnou myeloidnou leuk\u00e9miou(4). T\u00fdmito objavmi sa otvorila mo\u017enos\u0165 potenci\u00e1lneho vyu\u017eitia cirkuluj\u00facej n\u00e1dorovej DNA (ctDNA) ako molekul\u00e1rneho markera v diagnostike a monitorovan\u00ed n\u00e1dorov\u00fdch ochoren\u00ed.<\/p>\n

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P\u00f4vod cirkuluj\u00facej n\u00e1dorovej DNA<\/em><\/strong><\/p>\n

Zdrojom ctDNA v cirkul\u00e1cii onkologick\u00fdch pacientov s\u00fa mal\u00edgne bunky prim\u00e1rneho n\u00e1doru alebo n\u00e1dorov\u00fdch metast\u00e1z (obr\u00e1zok 1)<\/em><\/strong>. Podobne ako v pr\u00edpade cirkuluj\u00facich n\u00e1dorov\u00fdch buniek (CTCs) aj ctDNA sa do obehu m\u00f4\u017ee dosta\u0165 bu\u010f pas\u00edvnym, alebo akt\u00edvnym sp\u00f4sobom. Pas\u00edvny sp\u00f4sob uvo\u013e\u0148ovania ctDNA sa sp\u00e1ja s apopt\u00f3zou a nekr\u00f3zou n\u00e1dorov\u00fdch buniek, ktor\u00e9 boli fagocytovan\u00e9 makrof\u00e1gmi a in\u00fdmi \u010distiacimi bunkami(5).<\/p>\n

V pr\u00edpade akt\u00edvneho uvo\u013e\u0148ovania boli publikovan\u00e9 viacer\u00e9 \u0161t\u00fadie s cie\u013eom objasni\u0165, pre\u010do \u017eivotaschopn\u00e9 n\u00e1dorov\u00e9 bunky akt\u00edvne uvo\u013e\u0148uj\u00fa DNA do cirkul\u00e1cie. D\u00f4vodom je pravdepodobne schopnos\u0165 n\u00e1dorov\u00fdch buniek ovplyvni\u0165 transform\u00e1ciu vn\u00edmav\u00fdch buniek na vzdialen\u00fdch miestach v procese genometast\u00e1zy(6,7).<\/p>\n

\u010eal\u0161\u00edm zdrojom ctDNA s\u00fa CTCs pr\u00edtomn\u00e9 v krvi, av\u0161ak v po\u010dte CTCs a mno\u017estve ctDNA v krvi je zna\u010dn\u00fd nes\u00falad. Preto\u017ee jedna \u013eudsk\u00e1 bunka obsahuje 6 pg DNA a mililiter plazmy onkologick\u00fdch pacientov obsahuje priemerne 17 ng DNA(8), bolo by potrebn\u00fdch viac ako 2 000 CTCs na ml plazmy v pr\u00edpade, \u017ee by CTCs boli prim\u00e1rnym zdrojom ctDNA. V zmysle publikovan\u00fdch \u0161t\u00fadi\u00ed v\u0161ak 10 ml krvi obsahuje\u00a0 1 \u2013 10 CTCs u v\u00e4\u010d\u0161iny onkologick\u00fdch pacientov(9).<\/p>\n

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Vlastnosti cirkuluj\u00facej n\u00e1dorovej DNA<\/h3>\n

DNA v cirkul\u00e1cii je dvojvl\u00e1knov\u00e1 a fragmentovan\u00e1 s v\u00fdrazne ni\u017e\u0161ou molekulovou hmotnos\u0165ou v porovnan\u00ed s genomickou DNA. D\u013a\u017eka fragmentov v krvi cirkuluj\u00facej vo\u013enej DNA (cfDNA) sa pohybuje medzi kr\u00e1tkymi fragmentmi s d\u013a\u017ekou 70 \u2013 200 bp a fragmentmi dlh\u00fdmi pribli\u017ene 21 kilob\u00e1z(11) v z\u00e1vislosti od toho, ak\u00fdm mechanizmom bola cirkuluj\u00faca DNA uvo\u013enen\u00e1 do obehu. Kr\u00e1tke fragmenty poch\u00e1dzaj\u00fa z buniek, ktor\u00e9 pre\u0161li procesom apopt\u00f3zy. Ide o programovan\u00fa bunkov\u00fa smr\u0165, po\u010das ktorej je DNA \u0161tiepen\u00e1 nukle\u00e1zami na \u0161pecifick\u00fdch miestach, medzi nukleoz\u00f3mami. V s\u00falade s apoptotick\u00fdm p\u00f4vodom a d\u013a\u017ekou vl\u00e1kna DNA ovinut\u00e9ho okolo nukleoz\u00f3mu vr\u00e1tane spojovac\u00edch oblast\u00ed je distrib\u00facia d\u013a\u017eok cfDNA fragmentov preva\u017ene vo ve\u013ekosti 166 bp (alebo jej n\u00e1sobkov) (obr\u00e1zok 2)<\/em><\/strong>. Viacero \u0161t\u00fadi\u00ed preuk\u00e1zalo, \u017ee DNA pr\u00edtomn\u00e1 v plazme vykazuje o\u010dak\u00e1van\u00fd vzor fragment\u00e1cie (vzor \u201erebr\u00edka\u201c), pripom\u00ednaj\u00faci DNA \u0161tiepen\u00fa nukle\u00e1zami v miestach medzi nukleoz\u00f3mami, ktor\u00fd je typick\u00fd pr\u00e1ve pre apoptick\u00e9 bunky(11,12). Naopak, nekr\u00f3za je nekontrolovan\u00e1 bunkov\u00e1 smr\u0165, v d\u00f4sledku ktorej s\u00fa komponenty bunky nekontrolovane uvo\u013e\u0148ovan\u00e9 do prostredia bunky, \u010do vedie k po\u0161kodeniu bunky, a t\u00fdm aj k uvo\u013eneniu DNA fragmentov do\u00a0cirkul\u00e1cie. Tie s\u00fa v\u0161ak v\u00fdrazne dlh\u0161ie v porovnan\u00ed s fragmentmi poch\u00e1dzaj\u00facimi z apoptick\u00fdch buniek. V ned\u00e1vno publikovanej \u0161t\u00fadii zameranej na ur\u010denie d\u013a\u017eky fragmentov n\u00e1dorovej DNA autori Underhill a kol. preuk\u00e1zali na anim\u00e1lnom modeli \u013eudsk\u00e9ho glioblast\u00f3mu a hepatocelul\u00e1rneho karcin\u00f3mu, \u017ee naj\u010dastej\u0161ie zast\u00fapen\u00e9 fragmenty ctDNA s\u00fa dlh\u00e9 134 a 144 bp, zatia\u013e \u010do v norm\u00e1lnej cfDNA bol naj\u010dastej\u0161ie pr\u00edtomn\u00fd 167 bp dlh\u00fd fragment. Tieto zistenia boli zopakovan\u00e9 u pacientov s melan\u00f3mom. Navy\u0161e autori pr\u00e1ce uk\u00e1zali, \u017ee frekvencia mutovanej alely u pacientov s rakovinou p\u013e\u00fac bola vy\u0161\u0161ia vo frakcii, ktor\u00e1 obsahovala krat\u0161ie DNA fragmenty, najm\u00e4 ak distrib\u00facia fragmentov cirkuluj\u00facej DNA u onkologick\u00fdch pacientov bola podobn\u00e1 distrib\u00facii DNA fragmentov u zdrav\u00fdch kontrol. Okrem toho pozorovali aj ~10 bp periodicitu v d\u013a\u017eke fragmentov, ktor\u00e1 je ve\u013emi dobre op\u00edsan\u00e1 v s\u00favislosti s nukle\u00e1zovou aktivitou(13), \u010do je v s\u00falade s predch\u00e1- dzaj\u00facimi zisteniami, \u017ee ctDNA aj cfDNA s\u00fa spojen\u00e9 najm\u00e4 s apopt\u00f3zou buniek(11).<\/p>\n

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Koncentr\u00e1cia cirkuluj\u00facej n\u00e1dorovej DNA<\/h3>\n

Za norm\u00e1lnych fyziologick\u00fdch podmienok je cfDNA v perif\u00e9rnej krvi degradovan\u00e1 DN\u00e1zovou aktivitou, \u010do u zdrav\u00fdch jedincov vedie k n\u00edzkej koncentr\u00e1cii cfDNA (menej ako 1 \u2013 100 ng\/ml plazmy).V pr\u00edpade onkologick\u00fdch pacientov je DN\u00e1zov\u00e1 aktivita v ich krvnej plazme v porovnan\u00ed so zdrav\u00fdmi jedincami zn\u00ed\u017een\u00e1, \u010do u nich vedie k zv\u00fd\u0161enej koncentr\u00e1cii cirkuluj\u00facich nukleov\u00fdch kysel\u00edn (celkovej cfDNA obsahuj\u00facej aj ctDNA) (viac ako 1 000 ng\/ml plazmy)(11,15). Okrem toho sa predpoklad\u00e1, \u017ee u onkologick\u00fdch pacientov je zv\u00fd\u0161en\u00e1 miera apopt\u00f3zy a nekr\u00f3zy samotn\u00fdch n\u00e1dorov\u00fdch buniek a okolit\u00fdch norm\u00e1lnych buniek a ako n\u00e1dor zv\u00e4\u010d\u0161uje svoj objem, zvy\u0161uje sa aj po\u010det apoptotick\u00fdch a nekrotick\u00fdch buniek v d\u00f4sledku zv\u00fd\u0161enej bunkovej premeny. Za norm\u00e1lnych podmienok s\u00fa zvy\u0161ky t\u00fdchto buniek odstra\u0148ovan\u00e9 v procese fagocyt\u00f3zy, av\u0161ak t\u00e1 v n\u00e1dorovej mase nefunguje efekt\u00edvne, \u010do m\u00e1 za n\u00e1sledok akumul\u00e1ciu bunkov\u00fdch zvy\u0161kov a ich n\u00e1sledn\u00e9 uvo\u013enenie do cirkul\u00e1cie. V\u00fdsledkom je nerovnov\u00e1ha medzi uvo\u013e\u0148ovan\u00edm a elimin\u00e1ciou cfDNA. Koncentr\u00e1cia cfDNA medzi jednotlivcami zna\u010dne var\u00edruje a men\u00ed sa v \u010dase, pri\u010dom onkologick\u00ed pacienti vykazuj\u00fa vysok\u00fa variabilitu v mno\u017estve celkovej cfDNA a ctDNA frakcie, ktor\u00e1 sa pohybuje v rozmedz\u00ed od menej ako 1 % a\u017e do viac ako 90 % v z\u00e1vislosti od po\u010dtu a ve\u013ekosti n\u00e1dorov\u00fdch lo\u017e\u00edsk, prolifera\u010dn\u00e9ho \u0161t\u00e1dia, \u00farovne vaskulariz\u00e1cie, druhu a d\u013a\u017eky trvania lie\u010dby, odpovede na lie\u010dbu at\u010f.(14,16).<\/p>\n

Klinick\u00e9 vyu\u017eitie cirkuluj\u00facej n\u00e1dorovej DNA<\/h3>\n

CtDNA predstavuje do bud\u00facna jeden z najs\u013eubnej\u0161\u00edch n\u00e1strojov pre klinick\u00fa aplik\u00e1ciu, pri\u010dom m\u00f4\u017ee sl\u00fa\u017ei\u0165 ako biomarker progn\u00f3zy a recid\u00edvy n\u00e1dorov\u00fdch ochoren\u00ed, m\u00f4\u017ee indikova\u0165, \u010di je \u0161pecifick\u00e1 lie\u010dba v danom pr\u00edpade aplikovate\u013en\u00e1, alebo sa zn\u00ed\u017ei riziko recid\u00edvy a progresie n\u00e1dorov\u00e9ho ochorenia (obr\u00e1zok 3)<\/em><\/strong>.<\/p>\n

CtDNA ako diagnostick\u00fd biomarker. <\/strong>Jedn\u00fdmi z prv\u00fdch a najviac \u0161tudovan\u00fdch genetick\u00fdch zmien vo vz\u0165ahu k n\u00e1dorov\u00fdm ochoreniam s\u00fa mut\u00e1cie KRAS <\/em>g\u00e9nu. KRAS <\/em>je protoonkog\u00e9n z rodiny Ras <\/em>g\u00e9nov a mut\u00e1cie v tomto g\u00e9ne ved\u00fa ku kon\u0161titut\u00edvnej aktiv\u00e1cii EGFR signaliza\u010dnej dr\u00e1hy, ktor\u00e1 vedie k nekontrolovate\u013enej bunkovej prolifer\u00e1cii. Mut\u00e1cie\u00a0 v tomto g\u00e9ne sa be\u017ene vyskytuj\u00fa u pacientov s rakovinou p\u013e\u00fac a gastrointestin\u00e1lneho traktu(17,18). Mut\u00e1cie KRAS <\/em>g\u00e9nu v plazme alebo s\u00e9re pacientov s\u00fa relat\u00edvne \u0161pecifick\u00fdm markerom, preto\u017ee s\u00fa detegovate\u013en\u00e9 s vysokou \u0161pecificitou a senzitivitou u pacientov s kolorekt\u00e1lnym karcin\u00f3mom(19) a bola preuk\u00e1zan\u00e1 83 % zhoda medzi v\u00fdsledkami zo vzoriek z plazmy a t\u00fdmi, ktor\u00e9 boli z\u00edskan\u00e9 anal\u00fdzou n\u00e1dorov\u00e9ho tkaniva(20). Navy\u0161e v pr\u00edpade niektor\u00fdch pacientov boli mut\u00e1cie v cirkuluj\u00facej DNA KRAS <\/em>g\u00e9nu pr\u00edtomn\u00e9 e\u0161te pred prejavom neoplastick\u00e9ho ochorenia, \u010do nazna\u010duje, \u017ee by tak\u00e9to mut\u00e1cie mohli predstavova\u0165 vysok\u00e9 riziko vyvinutia tumoru(21). Podobne KRAS <\/em>mut\u00e1cie boli detegovan\u00e9 aj u pacientov s rakovinou pankreasu 5 \u2013 14 mesiacov pred diagn\u00f3zou tumoru a nevyskytovali sa u pacientov s ben\u00edgnymi patol\u00f3giami pankreasu ani u zdrav\u00fdch os\u00f4b(22).<\/p>\n

Hladina expresie niektor\u00fdch k\u013e\u00fa\u010dov\u00fdch g\u00e9nov asociovan\u00fdch s rakovinou m\u00f4\u017ee by\u0165 zmenen\u00e1 nielen v d\u00f4sledku genetick\u00fdch zmien, ale aj v d\u00f4sledku epigenetick\u00fdch modifik\u00e1ci\u00ed. Metyl\u00e1cia CpG ostrov\u010dekov v prom\u00f3torovej oblasti tumor-supresorov\u00fdch g\u00e9nov (TSG) sp\u00fa\u0161\u0165a lok\u00e1lne utlmenie expresie g\u00e9nu. Nadmern\u00e1 metyl\u00e1cia tumor-supresorov\u00e9ho g\u00e9nu p16 <\/em>bola n\u00e1jden\u00e1 v s\u00e9re pacientov s rakovinou pe\u010dene(23), p\u013e\u00fac(24) a prsn\u00edka(25). Metyl\u00e1cia TSG predstavuje jednu z prvotn\u00fdch pr\u00ed\u010din formovania tumoru, \u010d\u00edm sa otv\u00e1ra mo\u017e- nos\u0165 vyu\u017eitia anal\u00fdzy metyl\u00e1cie cirkuluj\u00facej DNA tumorov\u00e9ho p\u00f4vodu na skr\u00edning rakoviny v jej skorom \u0161t\u00e1diu. Fujiwara a kol. uk\u00e1zali, \u017ee detekcia statusu metyl\u00e1cie TSG v s\u00e9re je senzit\u00edvnej\u0161ia ako konven\u010dn\u00e9 markery pou\u017e\u00edvan\u00e9 na skor\u00fa diagnostiku rakoviny p\u013e\u00fac(26). Dokonca kombin\u00e1ciou cytologick\u00e9ho vy\u0161etrenia sp\u00fata a detekciou metyla\u010dn\u00e9ho statusu p16 <\/em>g\u00e9nu v plazme bolo mo\u017en\u00e9 identifikova\u0165 92 % pacientov s rakovinou p\u013e\u00fac(27).<\/p>\n

CtDNA ako prognostick\u00fd biomarker. <\/strong>Viacer\u00e9 \u0161t\u00fadie preuk\u00e1zali \u0161tatisticky signifikantn\u00fa korel\u00e1ciu medzi \u0161t\u00e1diom ochorenia a pr\u00edtomnos\u0165ou n\u00e1dorovo asociovan\u00fdch genetick\u00fdch zmien v krvi pacientov s n\u00e1dorom prsn\u00edka, vaje\u010dn\u00edkov, pankreasu a kolorekt\u00e1lnym karcin\u00f3mom. U pacientov\u00a0s kolorekt\u00e1lnym karcin\u00f3mom boli realizovan\u00e9 \u0161t\u00fadie zameran\u00e9 na detekciu hotspot <\/em>mut\u00e1ci\u00ed v KRAS <\/em>g\u00e9ne a hypermetyl\u00e1ciu prom\u00f3tora CDKN2A <\/em>(kin\u00e1zov\u00fd inhib\u00edtor 2 A dependentn\u00fd od cykl\u00ednu). Zistilo sa, \u017ee u 100 % pacientov s kolorekt\u00e1lnym karcin\u00f3mom (CRC), bez detekcie KRAS <\/em>mut\u00e1ci\u00ed a hypermetyl\u00e1cie prom\u00f3tora CDKN2A <\/em>v ctDNA, bola miera pre\u017eitia dva roky. Tieto v\u00fdsledky nazna\u010duj\u00fa, \u017ee ctDNA pr\u00edtomn\u00e1 v plazme je relevantn\u00fdm prognostick\u00fdm markerom v pr\u00edpade pacientov s CRC a tak\u00fdto marker m\u00f4\u017ee by\u0165 pou\u017eit\u00fd na identifik\u00e1ciu pacientov, u ktor\u00fdch je vysok\u00e9 riziko recid\u00edvy(28). Tieto v\u00fdsledky boli potvrden\u00e9 v \u010fal\u0161ej \u0161t\u00fadii, ke\u010f u pacientov s detegovate\u013enou ctDNA v plazme nastala recid\u00edva ochorenia v priebehu jedn\u00e9ho roka po odstr\u00e1nen\u00ed n\u00e1doru. Navy\u0161e vysok\u00e1 koncentr\u00e1cia cfDNA a detekcia KRAS <\/em>mut\u00e1ci\u00ed s\u00fa jasn\u00fdm ukazovate\u013eom nepriaznivej progn\u00f3zy u pacientov s metastatick\u00fdm CRC(29). Podobn\u00e9 v\u00fdsledky boli z\u00edskan\u00e9 aj v pr\u00edpade pacientov s rakovinou prsn\u00edka(30). Na z\u00e1klade multivaria\u010dnej anal\u00fdzy pr\u00edtomnos\u0165 KRAS <\/em>mut\u00e1ci\u00ed v plazme 256 pacientov s nemalobunkov\u00fdm karcin\u00f3mom p\u013e\u00fac (NSCLC) v pokro\u010dilom \u0161t\u00e1diu korelovala so zlou progn\u00f3zou u pacientov dost\u00e1vaj\u00facich prv\u00fa l\u00edniu chemoterapie. Pr\u00edtomnos\u0165 KRAS <\/em>mut\u00e1ci\u00ed v plazme bola vyhodnoten\u00e1 aj v s\u00fabore 44 pacientov s rakovinou pankreasu s cie\u013eom ur\u010di\u0165 progn\u00f3zu ochorenia u t\u00fdchto pacientov. Napriek n\u00edzkej senzitivite testu (27 %) bolo preuk\u00e1zan\u00e9, \u017ee pacienti, u ktor\u00fdch bola detegovan\u00e1 pr\u00edtomnos\u0165 KRAS <\/em>mut\u00e1ci\u00ed, mali signifikantne zn\u00ed\u017een\u00fa pravdepodobnos\u0165 pre\u017e\u00edvania v porovnan\u00ed s pacientmi bez uveden\u00fdch mut\u00e1ci\u00ed (17 % vs 41 % v pr\u00edpade 6 mesiacov a 0 % vs 24 % v pr\u00edpade 12 mesiacov)(31). Detekcia n\u00e1vratu ochorenia. <\/strong>S\u013eubnou klinickou aplik\u00e1ciou tekutej biopsie je skor\u00e1 detekcia n\u00e1vratu ochorenia po lie\u010dbe. V tomto oh\u013eade bola d\u00f4le\u017eit\u00e1 \u0161t\u00fadia, v ktorej monitorovanie n\u00e1dorovo \u0161pecifick\u00fdch zmien (vr\u00e1tane APC<\/em>, TP53 <\/em>a KRAS<\/em>) u pacientov s CRC, ktor\u00ed podst\u00fapili oper\u00e1ciu, viedlo k identifik\u00e1cii n\u00e1vratu ochorenia s takmer stopercentnou senzitivitou a \u0161pecificitou. Na z\u00e1klade pretrv\u00e1vania tumor-asociovan\u00fdch genetick\u00fdch zmien v cfDNA bezprostredne po oper\u00e1cii boli identifikovan\u00e9 aj v\u0161etky pr\u00edpady ne\u00faplnej resekcie u pacientov s rezidu\u00e1lnou chorobou. N\u00e1sledn\u00e9 \u0161t\u00fadie preuk\u00e1zali konzistentn\u00fd vz\u0165ah medzi n\u00e1vratom ochorenia a op\u00e4tovn\u00fdm objaven\u00edm ur\u010dit\u00fdch n\u00e1dorov\u00fdch zmien vr\u00e1tane APC<\/em>, TP53\u00a0<\/em>a KRAS <\/em>mut\u00e1ci\u00ed, ako aj alelovej nerovnov\u00e1hy(32-34).<\/p>\n

Dormancia je be\u017en\u00fdm javom mnoh\u00fdch n\u00e1dorov\u00fdch ochoren\u00ed a nemo\u017eno ju detegova\u0165 \u0161tandardn\u00fdmi met\u00f3dami. Sledovanie tumorovo \u0161pecifick\u00fdch zmien v po\u010dte k\u00f3pi\u00ed genomick\u00fdch oblast\u00ed u 50 pacientov s rakovinou prsn\u00edka pred oper\u00e1ciou aj po oper\u00e1cii preuk\u00e1zalo, \u017ee tieto aber\u00e1cie m\u00f4\u017eu v plazme pretrv\u00e1va\u0165 a\u017e do 12 rokov po diagn\u00f3ze n\u00e1dorov\u00e9ho ochorenia(35). Predikcia odpovede na lie\u010dbu. <\/strong>V s\u00fa\u010dasnosti sa v\u00fdber cielenej terapie v klinickej praxi riadi pr\u00edtomnos\u0165ou alebo absenciou jedinej genetickej zmeny v n\u00e1dorovej DNA. Molekul\u00e1rna anal\u00fdza v\u0161ak b\u00fdva \u010dasto vykon\u00e1van\u00e1 na archivovan\u00fdch tkanivov\u00fdch vzork\u00e1ch n\u00e1dorov, \u010do nie je optim\u00e1lny pr\u00edstup, ke\u010f\u017ee nemus\u00ed odr\u00e1\u017ea\u0165 aktu\u00e1lne genetick\u00e9 pozadie ochorenia. V pr\u00edpadoch, ke\u010f nemo\u017eno vykona\u0165 nov\u00fa biopsiu, by ctDNA mohla sl\u00fa\u017ei\u0165 ako ide\u00e1lny biomarker aktu\u00e1lneho molekul\u00e1rneho stavu ochorenia. Napr\u00edklad pre mana\u017ement pacientov s rakovinou prsn\u00edka je rozhoduj\u00face vyhodnotenie HER2 statusu. Na detekciu HER2 amplifik\u00e1cie v ctDNA u pacientov s metastatick\u00fdm karcin\u00f3mom prsn\u00edka Gevensleben a kol. optimalizovali met\u00f3du digit\u00e1lnej PCR, pri\u010dom na v\u00fdvoj met\u00f3dy bola pou\u017eit\u00e1 skupina 65 pacientov a na valid\u00e1ciu in\u00e1 nez\u00e1visl\u00e1 skupina 58 pacientov. Pacienti s metastatick\u00fdm ochoren\u00edm, ktor\u00ed boli identifikovan\u00ed ako HER2 pozit\u00edvni, tak mohli profitova\u0165 z HER2 cielenej terapie trastuzumabom. Dokonca traja pacienti s metastatick\u00fdm karcin\u00f3mom prsn\u00edka zahrnut\u00ed do tejto \u0161t\u00fadie z\u00edskali pri recid\u00edve ochorenia pozit\u00edvny HER2 status napriek tomu, \u017ee boli predt\u00fdm na z\u00e1klade biopsie tkaniva klasifikovan\u00ed ako HER2 negat\u00edvni(36).<\/p>\n

Na predikciu odpovede na lie\u010dbu mo\u017eno pou\u017ei\u0165 aj detekciu zv\u00fd\u0161en\u00e9ho mno\u017estva k\u00f3pi\u00ed in\u00fdch g\u00e9nov, ako napr. MET <\/em>amplifik\u00e1ciu na predpovedanie odpovede na anti-EGFR terapiu v pr\u00edpade rakoviny p\u013e\u00fac(37). Ako jasn\u00fd indik\u00e1tor odpovede na lie\u010dbu cetuximabom a irinotekanom pri metastatickom kolorekt\u00e1lnom karcin\u00f3me je pr\u00edtomnos\u0165, resp. kvantifik\u00e1cia mut\u00e1ci\u00ed KRAS <\/em>v plazme, pri\u010dom zv\u00fd\u0161en\u00e9 mno\u017estvo koreluje so zlou progn\u00f3zou. U niektor\u00fdch pacientov s n\u00edzkou hladinou KRAS <\/em>mut\u00e1ci\u00ed v plazme mo\u017eno anti-EGFR terapiou dosiahnu\u0165 ur\u010dit\u00fa \u00farove\u0148 stabiliz\u00e1cie ochorenia(29). Podobn\u00e9 v\u00fdsledky boli op\u00edsan\u00e9 aj v pr\u00edpade 64 pacientov s metastatick\u00fdm kolorekt\u00e1lnym karcin\u00f3mom, u ktor\u00fdch bol na lie\u010dbu pou\u017eit\u00fd temsirolimus a irinotekan bu\u010f samostatne, alebo v kombin\u00e1cii. Pacienti so zv\u00fd\u0161en\u00fdm podielom KRAS <\/em>mut\u00e1ci\u00ed v plazme lie\u010den\u00ed monoterapiou mali 77 % riziko skorej progresie ochorenia v porovnan\u00ed so 43 % rizikom v pr\u00edpade pacientov, u ktor\u00fdch bolo mno\u017estvo KRAS <\/em>mut\u00e1ci\u00ed v plazme n\u00edzke(38).<\/p>\n

Biomarker druhotnej rezistencie. <\/strong>V\u00fdvoj sekund\u00e1rnej (z\u00edskanej) rezistencie v d\u00f4sledku mut\u00e1cie je be\u017en\u00fdm d\u00f4sledkom cielenej terapie u v\u00e4\u010d\u0161iny pacientov s n\u00e1dorov\u00fdm ochoren\u00edm. Pr\u00edkladom je T790M substit\u00facia v membr\u00e1novom receptore EGFR, ktor\u00e1 m\u00e1 za n\u00e1sledok rezistenciu proti gefitinibu a erlotinibu asi u 50 % pacientov s rakovinou p\u013e\u00fac(39). Mut\u00e1cia bola najsk\u00f4r zisten\u00e1 u pacientov s relapsom a nesk\u00f4r potvrden\u00e1 neinvaz\u00edvnou anal\u00fdzou vzoriek plazmy, \u010d\u00edm bola potvrden\u00e1 mo\u017enos\u0165 vyu\u017eitia tekutej biopsie na monitorovanie rezistencie na cielen\u00fa terapiu(40,41).<\/p>\n

Podobne sekund\u00e1rna rezistencia proti anti-EGFR protil\u00e1tkam cetuximabu a panitumumabu, ktor\u00e9 sa vyu\u017e\u00edvaj\u00fa na lie\u010dbu kolorekt\u00e1lneho karcin\u00f3mu, je v\u00fdsledkom vzniku KRAS <\/em>mut\u00e1ci\u00ed v bunk\u00e1ch n\u00e1doru(42). Detekciou mut\u00e1ci\u00ed KRAS <\/em>v krvi pacientov po\u010das lie\u010dby cetuximabom a panitumumabom tak mo\u017eno detegova\u0165 vznik rezistentn\u00fdch alel a\u017e o 10 mesiacov sk\u00f4r ako so \u0161tandardn\u00fdmi met\u00f3dami(42,43). Diaz a kol. zistili v s\u00e9re 9 z 24 pacientov s CRC, ktor\u00fdch n\u00e1dory boli spo\u010diatku bez KRAS <\/em>mut\u00e1ci\u00ed, pr\u00edtomnos\u0165 detegovate\u013en\u00fdch mut\u00e1ci\u00ed KRAS <\/em>g\u00e9nu, pri\u010dom pri troch z nich boli detegovan\u00e9 viacer\u00e9 rozdielne mut\u00e1cie. V\u00fdskyt t\u00fdchto mut\u00e1ci\u00ed bol ve\u013emi konzistentn\u00fd, v\u0161eobecne nastal p\u00e4\u0165 a\u017e \u0161es\u0165 mesiacov po terapii(43).<\/p>\n

 <\/p>\n

Z\u00e1ver<\/h2>\n

Na z\u00e1ver m\u00f4\u017eeme skon\u0161tatova\u0165, \u017ee detekcia n\u00e1dorovo \u0161pecifick\u00fdch zmien vych\u00e1dzaj\u00faca z anal\u00fdzy cfDNA, resp. ctDNA, predstavuje obrovsk\u00fd pr\u00ednos v diagnostike a prognostike onkologick\u00fdch ochoren\u00ed. S\u00fa z\u00e1rove\u0148 n\u00e1dejnou cestou aj na zlep\u0161enie ich lie\u010dby nielen v\u010faka tomu, \u017ee predstavuj\u00fa neinvaz\u00edvnu alternat\u00edvu v\u00fdrazne menej za\u0165a\u017euj\u00facu pacienta aj zdravotn\u00e9 pracovisko pri odbere vzoriek, ale i preto, \u017ee maj\u00fa mo\u017enos\u0165 lep\u0161ie charakterizova\u0165 aktu\u00e1lny priebeh n\u00e1dorov\u00e9ho ochorenia, ktor\u00e9ho heterogenita je \u0161tandardn\u00fdmi postupmi identifikovate\u013en\u00e1 len vo ve\u013emi obmedzenom rozsahu. V bud\u00facnosti mo\u017eno po\u010d\u00edta\u0165 nielen s t\u00fdm, \u017ee anal\u00fdza cfDNA, resp. ctDNA bude doplnkom ku klasickej histopatologickej anal\u00fdze n\u00e1dorov\u00e9ho tkaniva, ale \u017ee v pr\u00edpade, \u017ee bude mo\u017en\u00e9 jej potenci\u00e1l vyu\u017ei\u0165 naplno, sa, naopak, histopatologick\u00e1 anal\u00fdza stane doplnkovou anal\u00fdzou k prim\u00e1rnej anal\u00fdze cfDNA, resp. ctDNA.<\/p>\n

Grantov\u00e1 podpora: <\/em><\/strong>T\u00e1to<\/em> publik\u00e1cia vznikla <\/em>s podporou gran<\/em>tov Agent\u00fary na podporu v\u00fdskumu a v\u00fdvoja \u2013 APVV-14-0273\u00a0 a APVV-14-0327.<\/em><\/p>\n

 <\/p>\n

 <\/p>\n

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

    \n
  1. Leon SA, Shapiro B, Sklaroff DM, et Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res 1977; 37(3): 646-650.<\/li>\n
  2. Stroun M, Anker P, Maurice P, et Neoplastic characteristics of the DNA found in the plasma of cancer patients. Oncology 1989; 46(5): 318-322.<\/li>\n
  3. Sorenson GD, Pribish DM, Valone FH, et Soluble normal and mutated DNA sequences from single-copy genes in human blood. Cancer Epidemiol Biomarkers Prev 1994; 3(1): 67-71.<\/li>\n
  4. Vasioukhin V, Anker P, Maurice P, et Point mutations of the N-ras gene in the blood plasma DNA of patients with myelodysplastic syndrome or acute myelogenous leukaemia. Br J Haematol 1994; 86(4): 774-779.<\/li>\n
  5. Diehl F, Li M, Dressman D, et Detection and quantification of mutations in the plasma of patients with colorectal tumors. Proc Natl Acad Sci USA 2005; 102(45): 16368-16373.<\/li>\n
  6. Garcia-Olmo DC, Dominguez C, Garcia-Arranz M, et Cell-free nucleic ac- ids circulating in the plasma of colorectal cancer patients induce the oncogenic transformation of susceptible cultured cells. Cancer Res 2010; 70(2): 560-567.<\/li>\n
  7. Trejo-Becerril C, Perez-Cardenas E, Taja-Chayeb L, et Cancer progression mediated by horizontal gene transfer in an in vivo model. PLoS One 2012; 7(12): e52754.<\/li>\n
  8. Perkins G, Yap TA, Pope L, et Multi-purpose utility of circulating plasma DNA testing in patients with advanced cancers. PLoS One 2012; 7(11): e47020.<\/li>\n
  9. Alix-Panabieres C, Pantel K. Challenges in circulating tumour cell research. Nat Rev Cancer 2014; 14(9): 623-631.<\/li>\n
  10. Schwarzenbach H, Hoon DS, Pantel Cell-free nucleic acids as biomark- ers in cancer patients. Nat Rev Cancer 2011; 11(6): 426-437.<\/li>\n
  11. Jahr S, Hentze H, Englisch S, et DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. Cancer Res 2001; 61(4): 1659-1665.<\/li>\n
  12. Ellinger J, Wittkamp V, Albers P, et Cell-free circulating DNA: diagnostic value in patients with testicular germ cell cancer. J Urol 2009; 181(1): 363-371.<\/li>\n
  13. Underhill HR, Kitzman JO, Hellwig S, et Fragment Length of Circulating Tumor DNA. PLoS Genet 2016; 12(7): e1006162.<\/li>\n
  14. Heitzer E, Ulz P, Geigl Circulating tumor DNA as a liquid biopsy for cancer. Clin Chem 2015; 61(1): 112-123.<\/li>\n
  15. Mouliere F, Robert B, Arnau Peyrotte E, et al. High fragmentation characterizes tumour-derived circulating PLoS One 2011; 6(9): e23418.<\/li>\n
  16. Bettegowda C, Sausen M, Leary RJ, et al. Detection of circulating tu- mor DNA in early- and late-stage human Sci Transl Med 2014; 6(224): 224ra224.<\/li>\n
  17. Vogelstein B, Fearon ER, Hamilton SR, et al. Genetic alterations during colorectal-tumor N Engl J Med 1988; 319(9): 525-532.<\/li>\n
  18. Slebos RJ, Kibbelaar RE, Dalesio O, et al. K-ras oncogene activation as a prognostic marker in adenocarcinoma of the lung. N Engl J Med 1990; 323(9): 561-565.<\/li>\n
  19. Sorenson GD, Pribish DM, Valone FH, et Soluble normal and mutated DNA sequences from single-copy genes in human blood. Cancer Epidemiol Biomarkers Prev 1994; 3(1): 67-71.<\/li>\n
  20. Kopreski MS, Benko FA, Kwee C, et Detection of mutant K-ras DNA in plasma or serum of patients with colorectal cancer. Br J Cancer 1997; 76(10): 1293-1299.<\/li>\n
  21. Kopreski MS, Benko FA, Borys DJ, et Somatic mutation screening: identification of individuals harboring K-ras mutations with the use of plasma DNA. J Natl Cancer Inst 2000; 92(11): 918-923.<\/li>\n
  22. Mulcahy HE, Lyautey J, Lederrey C, et A prospective study of K-ras mutations in the plasma of pancreatic cancer patients. Clin Cancer Res 1998; 4(2): 271-275.<\/li>\n
  23. Wong IH, Lo YM, Zhang J, et Detection of aberrant p16 methylation in the plasma and serum of liver cancer patients. Cancer Res 1999; 59(1): 71-73.<\/li>\n
  24. Esteller M, Sanchez-Cespedes M, Rosell R, et Detection of aberrant promoter hypermethylation of tumor suppressor genes in serum DNA from non- small cell lung cancer patients. Cancer Res 1999; 59(1): 67-70.<\/li>\n
  25. Silva JM, Dominguez G, Villanueva MJ, et Aberrant DNA methylation of the p16INK4a gene in plasma DNA of breast cancer patients. Br J Cancer 1999; 80(8): 1262-1264.<\/li>\n
  26. Fujiwara K, Fujimoto N, Tabata M, et Identification of epigenetic aberrant promoter methylation in serum DNA is useful for early detection of lung cancer. Clin Cancer Res 2005; 11(3): 1219-1225.<\/li>\n
  27. Liu Y, An Q, Li L, et Hypermethylation of p16INK4a in Chinese lung cancer patients: biological and clinical implications. Carcinogenesis 2003; 24(12):1897-1901.<\/li>\n
  28. Lecomte T, Berger A, Zinzindohoue F, et Detection of free-circulating tumor-associated DNA in plasma of colorectal cancer patients and its asso- ciation with prognosis. Int J Cancer 2002; 100(5): 542-548.<\/li>\n
  29. Spindler KL, Pallisgaard N, Vogelius I, et Quantitative cell-free DNA, KRAS, and BRAF mutations in plasma from patients with metastatic colorec- tal cancer during treatment with cetuximab and irinotecan. Clin Cancer Res 2012; 18(4): 1177-1185.<\/li>\n
  30. Dawson SJ, Tsui DW, Murtaza M, et Analysis of circulating tumor DNA to monitor metastatic breast cancer. N Engl J Med 2013; 368(13): 1199-1209.<\/li>\n
  31. Castells A, Puig P, Mora J, et K-ras mutations in DNA extracted from the plasma of patients with pancreatic carcinoma: diagnostic utility and prognostic significance. J Clin Oncol 1999; 17(2): 578-584.<\/li>\n
  32. Frattini M, Gallino G, Signoroni S, et Quantitative and qualitative characterization of plasma DNA identifies primary and recurrent colorectal can- cer. Cancer Lett 2008; 263(2): 170-181.<\/li>\n
  33. Hamana K, Uzawa K, Ogawara K, Shiiba M, Bukawa H, Yokoe H, Tanzawa<\/li>\n
  34. Monitoring of circulating tumour-associated DNA as a prognostic tool for oral squamous cell carcinoma. Br J Cancer 2005; 92(12): 2181-2184.<\/li>\n
  35. Hsieh JS, Lin SR, Chang MY, et APC, K-ras, and p53 gene mutations in colorectal cancer patients: correlation to clinicopathologic features and post- operative surveillance. Am Surg 2005; 71(4): 336-343.<\/li>\n
  36. Shaw JA, Page K, Blighe K, et al. Genomic analysis of circulating cell-free DNA infers breast cancer Genome Res 2012; 22(2): 220-231.<\/li>\n
  37. Gevensleben H, Garcia-Murillas I, Graeser MK, et Noninvasive detection of HER2 amplification with plasma DNA digital PCR. Clin Cancer Res 2013; 19(12): 3276-3284.<\/li>\n
  38. Bean J, Brennan C, Shih JY, et MET amplification occurs with or without T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib. Proc Natl Acad Sci USA 2007; 104(52): 20932-20937.<\/li>\n
  39. Spindler KL, Sorensen MM, Pallisgaard N, et Phase II trial of temsirolimus alone and in combination with irinotecan for KRAS mutant metastatic colorectal cancer: outcome and results of KRAS mutational analysis in plasma. Acta Oncol 2013; 52(5): 963-970.<\/li>\n
  40. Kosaka T, Yatabe Y, Endoh H, et Analysis of epidermal growth factor receptor gene mutation in patients with non-small cell lung cancer and acquired resistance to gefitinib. Clin Cancer Res 2006; 12(19): 5764-5769.<\/li>\n
  41. Nakamura T, Sueoka-Aragane N, Iwanaga K, et Application of a highly sensitive detection system for epidermal growth factor receptor mutations in plasma DNA. J Thorac Oncol 2012; 7(9): 1369-1381.<\/li>\n
  42. Taniguchi K, Uchida J, Nishino K, et al. Quantitative detection of EGFR mutations in circulating tumor DNA derived from lung Clin Cancer Res 2011; 17(24): 7808-7815.<\/li>\n
  43. Misale S, Yaeger R, Hobor S, et al. Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal Nature 2012; 486(7404): 532-536.<\/li>\n
  44. Diaz LA, Jr., Williams RT, Wu J, et al. The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers. Nature 2012; 486(7404): 537-540.<\/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.   Cirkuluj\u00faca n\u00e1dorov\u00e1 DNA Zv\u00fd\u0161en\u00e1 hladina cirkuluj\u00facich nukleov\u00fdch kysel\u00edn u pacientov s rakovinou bola prv\u00fdkr\u00e1t op\u00edsan\u00e1 u\u017e v roku 1977(1). Trvalo \u010fal\u0161\u00edch 10 rokov, pok\u00fdm Stroun a kol. preuk\u00e1zali pr\u00edtomnos\u0165<\/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":[608,611,900,1031],"class_list":["post-1582","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-circulating-dna","tag-circulating-tumor-dna","tag-liquid-biopsy","tag-oncological-diseases","typ_clanku-review-article"],"acf":{"abstrakt":"

    Identification of the presence of tumor DNA in circulation of oncological patients has brought new possibilities for their use in clinical-diagnostic practice. Following the utilization of new methods, that enabled highly sensitive and sufficiently specific detection of circulating tumor DNA, many studies have been published demonstrating the vast potential of detection and quantification of circulating tumor DNA in clinical management of oncological patients. That is why we have seen enormous efforts in recent years to implement results of these studies into routine clinical practice and, according to current knowledge, we can look forward for their use in early diagnosis, prognosis, early detection of disease relapse, or as a biomarker for the selection of clinical therapy all in so-called non-invasive manner, in the form of liquid biopsy.<\/p>\n

    Keywords:<\/strong> circulating DNA, circulating tumor DNA, oncological diseases, liquid biopsy<\/p>\n","casopis":[{"ID":1513,"post_author":"7","post_date":"2018-11-05 11:53:53","post_date_gmt":"2018-11-05 10:53:53","post_content":"

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    • Circulating tumor DNA and its utilization as marker with prognostic, predictive and diagnostic value in patients with oncological diseases<\/li>\r\n<\/ul>","post_title":"newsLab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-2","to_ping":"","pinged":"","post_modified":"2018-11-05 11:57:18","post_modified_gmt":"2018-11-05 10:57:18","post_content_filtered":"","post_parent":0,"guid":"http:\/\/www.newslab.sk\/casopis\/newslab-2\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"102","upload_clanok":{"ID":1580,"id":1580,"title":"Cirkuluj\u00faca n\u00e1dorov\u00e1 DNA a jej vyu\u017eitie ako markera s prognostickou,","filename":"Cirkuluj\u00faca-n\u00e1dorov\u00e1-DNA-a-jej-vyu\u017eitie-ako-markera-s-prognostickou.pdf","filesize":423043,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2018\/11\/Cirkuluj\u00faca-n\u00e1dorov\u00e1-DNA-a-jej-vyu\u017eitie-ako-markera-s-prognostickou.pdf","link":"https:\/\/www.newslab.sk\/en\/cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-prediktivnou-a-diagnostickou-hodnotou-u-pacientov-s-nadorovymi-ochoreniami\/cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-2\/","alt":"","author":"7","description":"","caption":"","name":"cirkulujuca-nadorova-dna-a-jej-vyuzitie-ako-markera-s-prognostickou-2","status":"inherit","uploaded_to":1582,"date":"2018-11-11 13:30:51","modified":"2018-11-11 13:30:51","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\/1582","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=1582"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1582\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1582"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1582"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1582"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}