{"id":1011,"date":"2017-04-06T20:18:08","date_gmt":"2017-04-06T18:18:08","guid":{"rendered":"http:\/\/www.newslab.sk\/2017\/04\/06\/cirkulujuca-dna-od-jej-objavenia-cez-neinvazivne-prenatalne-testovanie-aneuploidii-a-monogenne-dedicnych-ochoreni-az-k-neinvazivnej-nadorovej-diagnostike-a-prognostike\/"},"modified":"2017-10-03T07:53:40","modified_gmt":"2017-10-03T05:53:40","slug":"circulating-dna-from-its-discovery-to-non-invasive-prenatal-testing-of-aneuploidies-and-monogenic-hereditary-diseases-to-non-invasive-cancer-diagnosis-and-prognostics","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/circulating-dna-from-its-discovery-to-non-invasive-prenatal-testing-of-aneuploidies-and-monogenic-hereditary-diseases-to-non-invasive-cancer-diagnosis-and-prognostics\/","title":{"rendered":"Circulating DNA \u2013 from its discovery to non-invasive prenatal testing of aneuploidies and monogenic hereditary diseases to non-invasive cancer diagnosis and prognostics"},"content":{"rendered":"<pre><span style=\"color: #ff0000;\"><strong>*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf <\/strong><\/span>\r\n <span style=\"color: #ff0000;\"><strong>attachment at the end of the paper.<\/strong> <\/span><\/pre>\n<p>&nbsp;<\/p>\n<p><strong>\u00dav<\/strong><strong>od<\/strong><\/p>\n<p>D\u00f4le\u017eitos\u0165 objavenia cirkuluj\u00facej DNA v roku 1947 Mandelom a Metaisom bola \u00a0dlh\u00fd \u010das \u00a0nerozpoznan\u00e1(1). A\u017e o dvadsa\u0165 rokov, v roku 1966, \u00a0bola \u00a0publikovan\u00e1 \u0161t\u00fadia poukazuj\u00faca na asoci\u00e1ciu medzi \u00a0zv\u00fd\u0161enou hladinou cirkuluj\u00facej \u00a0DNA v s\u00e9re pacientov so syst\u00e9mov\u00fdm lupus \u00a0erythematosus(2). V roku 1973 \u00a0bola zv\u00fd\u0161en\u00e1 hladina \u00a0cirkuluj\u00facej \u00a0DNA identifikovan\u00e1 \u00a0aj pri \u010fal\u0161\u00edch \u00a0z\u00e1va\u017en\u00fdch ochoreniach, napr. pri reumatoidnej artrit\u00edde, glomerulovej nefrit\u00edde, pankreatit\u00edde, z\u00e1palov\u00fdch ochoreniach \u010driev a hepatit\u00edde(3). Kr\u00e1tko nato, v roku 1977, \u00a0u\u017e bola \u00a0op\u00edsan\u00e1 asoci\u00e1cia aj medzi \u00a0zv\u00fd\u0161en\u00fdmi hladinami cirkuluj\u00facej \u00a0DNA a n\u00e1dorov\u00fdm ochoren\u00edm u pacientov s lymf\u00f3mom, \u00a0gli\u00f3mom, rakovinou \u00a0p\u013e\u00fac, maternice, kr\u010dka maternice, prsn\u00edka, \u00a0kolorekta, \u00a0v porovnan\u00ed so zdrav\u00fdmi jedincami a, navy\u0161e, \u00a0pacienti s lokalizovan\u00fdm n\u00e1dorom dosahovali ni\u017e\u0161iu koncentr\u00e1ciu cirkuluj\u00facej \u00a0DNA v porovnan\u00ed s pacient- mi s metastatick\u00fdm ochoren\u00edm. \u010co bolo e\u0161te d\u00f4le\u017eitej\u0161ie, autori \u0161t\u00fadie prv\u00fdkr\u00e1t \u00a0preuk\u00e1zali, \u017ee \u00a0po lie\u010dbe \u00a0t\u00fdchto \u00a0pacientov bo zaznamenan\u00fd pokles hladiny cirkuluj\u00facej DNA u \u010dasti pacientov, a \u017ee jej sledovanie m\u00f4\u017ee ma\u0165 potenci\u00e1l vyu\u017eitia v prognostike n\u00e1dorov\u00fdch ochoren\u00ed(4). V tom obdob\u00ed\u00a0 v\u0161ak neexistovali \u00a0dostato\u010dne senzit\u00edvne a \u0161pecifick\u00e9 met\u00f3dy, ktor\u00e9 by umo\u017e\u0148ovali identifikova\u0165 \u00a0n\u00e1dorovo \u0161pecifick\u00fa cirkuluj\u00facu DNA a kvantitat\u00edvne anal\u00fdzy boli realizovan\u00e9 prostredn\u00edctvom \u00a0biochemick\u00fdch met\u00f3d alebo \u00a0s vyu\u017eit\u00edm univerz\u00e1lnych genetick\u00fdch markerov (ako napr. <em>Alu<\/em>-PCR).<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Cirkuluj\u00faca <\/strong><strong>v<\/strong><strong>o\u013en\u00e1 fet\u00e1lna DNA \u2013 genetick\u00e1 \u00e9ra<\/strong><\/p>\n<p>\u010eal\u0161\u00edm\u00a0 d\u00f4le\u017eit\u00fdm m\u00ed\u013enikom vo v\u00fdskume cirkuluj\u00facej \u00a0DNA bol v roku 1989 \u00a0objav pr\u00edtomnosti tzv. cirkuluj\u00facej \u00a0vo\u013enej fet\u00e1lnej \u00a0DNA (cffDNA \u2013 cell free \u00a0fetal \u00a0DNA) ako \u00a0s\u00fa\u010dasti \u00a0celkovej cirkuluj\u00facej \u00a0DNA v krvi tehotn\u00fdch \u017eien, ktor\u00fd predznamenal \u00a0nielen \u00a0vznik a rozvoj novej oblasti tzv. neinvaz\u00edvneho prenat\u00e1lneho testovania (NIPT), ale z\u00e1rove\u0148 na\u0161tartoval vysok\u00fd z\u00e1ujem o oblas\u0165 \u00a0v\u00fdskumu \u00a0cirkuluj\u00facich nukleov\u00fdch \u00a0kysel\u00edn v\u0161eobecne(5). Prvou\u00a0 klinicky vyu\u017eite\u013enou aplik\u00e1ciou v\u00fdskumu bol test zalo\u017een\u00fd na kvalitat\u00edvnej \u00a0detekcii \u00a0konkr\u00e9tnych Y-chromoz\u00f3mov\u00fdch sekvenci\u00ed v matern\u00e1lnej plazme, umo\u017e\u0148uj\u00faci neinvaz\u00edvne ur\u010denie pohlavia \u00a0plodu z krvi tehotnej. Pr\u00edtomnos\u0165 t\u00fdchto \u00a0sekvenci\u00ed bola \u00a0na \u00fa\u010dely stanovenia pohlavia plodu krit\u00e9riom, ktor\u00e9 umo\u017enilo vyu\u017eitie anal\u00fdzy pri neinvaz\u00edvnom prenat\u00e1lnom testovan\u00ed X-viazan\u00fdch dedi\u010dn\u00fdch ochoren\u00ed. Po pridan\u00ed \u00a0kvantitat\u00edvneho rozmeru anal\u00fdzy \u00a0boli definovan\u00e9 mno\u017estv\u00e1 fet\u00e1lnej DNA v matern\u00e1lnej cirkul\u00e1cii, ke\u010f p\u00f4vodn\u00e1 pr\u00e1ca z roku 1998 \u00a0preuk\u00e1zala nielen pr\u00edtomnos\u0165 dostato\u010dn\u00e9ho mno\u017estva fet\u00e1lnej DNA pre r\u00f4zne \u00a0typy genetick\u00fdch anal\u00fdz u\u017e v skor\u00fdch \u00a0\u0161t\u00e1di\u00e1ch tehotenstva, ale aj n\u00e1rast podielu fet\u00e1lnej DNA s postupuj\u00facim tehotenstvom. Podiel \u00a0fet\u00e1lnej DNA nar\u00e1stol od 7. t\u00fd\u017ed\u0148a \u00a0tehotenstva z priemern\u00fdch 25 GEq\/ml (genomick\u00fdch ekvivalentov\/ml plazmy) \u00a0na priemern\u00fdch 300 GEq\/ml(6). \u00a0O rok nesk\u00f4r bolo \u00a0navy\u0161e preuk\u00e1zan\u00e9 r\u00fdchle \u00a0odb\u00faranie fet\u00e1lnej DNA po p\u00f4rode, ke\u010f pol\u010das rozpadu fet\u00e1lnej DNA bol na \u00farovni 16 min\u00fat \u00a0a prakticky \u00a0\u00fapln\u00e9 \u00a0odb\u00faranie fet\u00e1lnej DNA do dvoch hod\u00edn po p\u00f4rode(7). To znamenalo ve\u013ek\u00fa v\u00fdhodu v porovnan\u00ed s dovtedy \u00a0konkuren\u010dnou strat\u00e9giou neinvaz\u00edvneho prenat\u00e1lneho DNA testovania zalo\u017een\u00e9ho na anal\u00fdze cirkuluj\u00facich \u00a0fet\u00e1lnych buniek. Ich nev\u00fdhodou bolo na jednej strane ve\u013emi n\u00edzke zast\u00fapenie v krvi tehotnej \u2013 2 \u2013 20\/ml krvi(8,9), na druhej strane dlhodob\u00e9 pre\u017e\u00edvanie fet\u00e1lnych buniek v krvi matky (mikrochim\u00e9rizmus), ktor\u00e9 mohlo kompromitova\u0165 v\u00fdsledky DNA anal\u00fdzy buniek z odberu krvi v priebehu aktu\u00e1lnej tehotnosti s bunkami z tej predch\u00e1dzaj\u00facej(10).<\/p>\n<p>V nadv\u00e4znosti \u00a0na tieto \u00a0zistenia sa rozvoj v oblasti neinvaz\u00edvnej prenat\u00e1lnej DNA diagnostiky zameral pr\u00e1ve na cffDNA. V\u010faka tomu sa postupne do klinickej praxe dostali testy \u00a0pre determin\u00e1ciu RhD statusu plodu(11) \u010di detekciu monog\u00e9nov\u00fdch mendelistick\u00fdch \u00a0ochoren\u00ed s autozom\u00e1lne dominantnou dedi\u010dnos\u0165ou v pr\u00edpadoch ich dedi\u010dnosti od otca \u00a0plodu, \u00a0ako \u00a0napr. \u00a0achondropl\u00e1zia(12), myotonick\u00e1 dystrofia(13). V nadv\u00e4znosti na zistenie, \u017ee DNA fet\u00e1lneho a matern\u00e1lneho p\u00f4vodu \u00a0s\u00fa rozdielne fragmentovan\u00e9, pri\u010dom fet\u00e1lna DNA bola fragmentovan\u00e1 viac ako matern\u00e1lna(14), sa v nasleduj\u00facom obdob\u00ed za\u010dala testova\u0165 mo\u017enos\u0165 vyu\u017eitia frakcion\u00e1cie cirkuluj\u00facej \u00a0DNA pred \u00a0samotnou molekul\u00e1rnou anal\u00fdzou. T\u00e1 bola v za\u010diatkoch sprostredkovan\u00e1 predov\u0161etk\u00fdm vyrez\u00e1van\u00edm frakcie DNA s ve\u013ekos\u0165ou &lt;300 bp (b\u00e1zov\u00fdch p\u00e1rov) z agar\u00f3zov\u00fdch g\u00e9lov po elektrofor\u00e9ze izolovanej DNA. Takto predpripraven\u00e1 DNA vykazovala preukaznej\u0161ie v\u00fdsledky pri detekcii patog\u00e9nnych variantov v DNA plodu napr. pri achondropl\u00e1zii, \u03b2-talas\u00e9mii \u010di dokonca pri neinvaz\u00edvnej detekcii parent\u00e1lnych alel STR markerov vyu\u017e\u00edvan\u00fdch \u0161tandardne pri \u00a0detekcii chromoz\u00f3mov\u00fdch aneuploidi\u00ed(15-17). V tomto obdob\u00ed \u00a0boli detegovan\u00e9 a na \u00fa\u010dely vyu\u017eitia pri NIPT validovan\u00e9 konkr\u00e9tne oblasti DNA plodu (napr. RASSF1A), ktor\u00e9 boli v porovnan\u00ed s matern\u00e1lnou DNA rozdielne metylovan\u00e9 a mali umo\u017eni\u0165 zv\u00fd\u0161enie spo\u013eahlivosti NIPT anal\u00fdz najm\u00e4 v\u010faka presnej\u0161iemu stanoveniu podielu fet\u00e1lnej DNA vo vzorke(18). S n\u00e1stupom technol\u00f3gie digit\u00e1lnej PCR bola e\u0161te preuk\u00e1zan\u00e1 mo\u017enos\u0165 neinvaz\u00edvnej determin\u00e1cie pr\u00edtomnosti triz\u00f3mie chromoz\u00f3mu 21 u plodu v pr\u00edpade, \u017ee jeho fet\u00e1lna DNA bola zast\u00fapen\u00e1 v celkovej cirkuluj\u00facej DNA tehotnej nad 25 %(19). Ani t\u00e1to met\u00f3da v\u0161ak nebola v\u0161eobecne vyu\u017eite\u013en\u00e1 pre rutinn\u00e9 \u00a0pou\u017eitie NIPT pri detekcii chromoz\u00f3mov\u00fdch aneuploidi\u00ed plodu, preto\u017ee\u00a0 v tom \u010dase publikovan\u00e9 \u00fadaje \u00a0uv\u00e1dzali \u00a0priemern\u00fd podiel cffDNA v matern\u00e1lnej cirkul\u00e1cii maxim\u00e1lne na \u00farovni 12 %(6).<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Cirkuluj\u00faca <\/strong><strong>v<\/strong><strong>o\u013en\u00e1 fet\u00e1lna DNA \u2013 genomick\u00e1\u00e9ra<\/strong><\/p>\n<p>\u010eal\u0161\u00ed rozmach s\u00favisel s n\u00e1stupom \u00e9ry genomiky sprostredkovanej predov\u0161etk\u00fdm dostupnos\u0165ou mas\u00edvneho paraleln\u00e9ho sekvenovania (MPS). Technol\u00f3gia MPS sp\u00f4sobila v oblasti NIPT revol\u00faciu. \u00a0Od roku 2008, ke\u010f boli simult\u00e1nne publikovan\u00e9 \u0161t\u00fadie preukazuj\u00face potenci\u00e1l vyu\u017eitia MPS pri neinvaz\u00edvnej detekcii triz\u00f3mi\u00ed chromoz\u00f3mov 21, 18 a 13 <strong><em>(obr\u00e1zok 1)<\/em><\/strong>(20,21), sa rozbehol rozsiahly \u00a0v\u00fdskum, ktor\u00fd priniesol \u00a0nov\u00e9 \u00a0z\u00e1kladn\u00e9 inform\u00e1cie o charakteristike cffDNA v matern\u00e1lnej cirkul\u00e1cii. Okrem toho u\u017e v p\u00f4vodnej pr\u00e1ci Fan a kol., 2008(20) bol op\u00edsan\u00fd charakteristick\u00fd ve\u013ekostn\u00fd profil DNA fragmentov cirkuluj\u00facej DNA, ktor\u00fd pouk\u00e1zal na fragment\u00e1ciu zalo\u017een\u00fa na ve\u013ekosti DNA sekvencie zodpovedaj\u00facej nukleoz\u00f3mu. T\u00e1to \u00a0inform\u00e1cia bola nesk\u00f4r \u0161pecifikovan\u00e1 a bolo preuk\u00e1zan\u00e9, \u017ee priemern\u00e1 ve\u013ekos\u0165 DNA fragmentov fet\u00e1lneho p\u00f4vodu \u00a0je 143 bp, k\u00fdm fragmenty matern\u00e1lnej DNA maj\u00fa \u00a0priemern\u00fa ve\u013ekos\u0165 166 bp <strong><em>(obr\u00e1zok \u00a02)<\/em><\/strong>(22).<\/p>\n<p>Z\u00e1rove\u0148 \u00a0bola technol\u00f3gia MPS vyu\u017eit\u00e1 na pr\u00edpravu \u00a0univerz\u00e1lneho \u00a0postupu umo\u017e\u0148uj\u00faceho neinvaz\u00edvnym sp\u00f4sobom determin\u00e1ciu prenosu oboch parent\u00e1lnych alel z rodi\u010dov na plod, nielen pri autozom\u00e1lne dominantn\u00fdch ochoreniach deden\u00fdch po otcovi, ale aj pri autozom\u00e1lne reces\u00edvnych ochoreniach prostredn\u00edctvom kvantitat\u00edvnej genotypovej anal\u00fdzy cirkuluj\u00facej \u00a0DNA vych\u00e1dzaj\u00facej z digit\u00e1lne \u00a0ve\u013ekostne selektovan\u00fdch MPS d\u00e1t \u00a0a s vyu\u017eit\u00edm met\u00f3dy tzv. stanovenia relat\u00edvnej muta\u010dnej d\u00e1vky(23). Ve\u013ekostn\u00e1 charakteristika sl\u00fa\u017eila nesk\u00f4r pri optimaliz\u00e1cii met\u00f3d pr\u00edpravy vzoriek pre MPS anal\u00fdzy, ale aj pri \u00faprave \u00a0bioinformatick\u00fdch algoritmov vyu\u017e\u00edvan\u00fdch aj v s\u00fa\u010dasnosti pri NIPT zameranom na neinvaz\u00edvnu detekciu z\u00e1va\u017en\u00fdch triz\u00f3mi\u00ed plodu.<\/p>\n<p>Z\u00e1rove\u0148 \u00a0v tej istej pr\u00e1ci bola prostredn\u00edctvom pokro\u010dilej \u00a0haplotypovej anal\u00fdzy\u00a0 determinovan\u00e1 cel\u00e1 chromoz\u00f3mov\u00e1 mapa plodu vych\u00e1dzaj\u00faca z anal\u00fdzy cirkuluj\u00facej DNA tehotnej a DNA matky a otca \u00a0plodu, \u010do predznamenalo \u00e9ru neinvaz\u00edvneho sekvenovania gen\u00f3mu plodu s potenci\u00e1lnymi aplik\u00e1ciami, napr. aj v oblasti v\u00fdskumu\u00a0 cirkuluj\u00facich n\u00e1dorov\u00fdch DNA. V rokoch\u00a0 2010 \u2013 2013 boli v rozsiahlych \u0161t\u00fadi\u00e1ch validovan\u00e9 met\u00f3dy a pr\u00edstupy \u00a0k anal\u00fdze vzoriek tehotn\u00fdch \u017eien s cie\u013eom \u00a0detekcie triz\u00f3mi\u00ed chromoz\u00f3mov 21, 18 a 13, ktor\u00e9 \u00a0preuk\u00e1zali vysok\u00fa \u00a0senzitivitu (&gt; 99,9 % pre triz\u00f3miu 21; &gt; 91,9 % pre triz\u00f3miu \u00a018; &gt; 78,6 % pre triz\u00f3miu \u00a013; &gt; 93,8 % pre monoz\u00f3miu X) a \u0161pecificitu (&gt; 97,9 % pre autoz\u00f3mov\u00e9 aneuploidie), tak\u00e9hoto neinvaz\u00edvneho prenat\u00e1lneho testovania(24-27). Nasledovala publik\u00e1cia aktualizuj\u00faca a opisuj\u00faca priemern\u00fd podiel fet\u00e1lnej DNA (fet\u00e1lna frakcia) u tehotn\u00fdch v 11. t\u00fd\u017edni tehotenstva, \u010do je z poh\u013eadu na\u010dasovania NIPT ide\u00e1lny \u010das, v hodnote 10,2 % <strong><em>(obr\u00e1zok 3)<\/em><\/strong>(28). \u010eal\u0161ia preuk\u00e1zala medzipopula\u010dn\u00e9 rozdiely v priemernej fet\u00e1lnej frakcii a tie\u017e z\u00e1vislos\u0165 \u00a0fet\u00e1lnej frakcie \u00a0od hmotnosti tehotnej. Fet\u00e1lna frakcia bola ni\u017e\u0161ia u tehotn\u00fdch afrokaribsk\u00e9ho p\u00f4vodu \u00a0v porovnan\u00ed s tehotn\u00fdmi kaukazsk\u00e9ho p\u00f4vodu \u00a0a k\u00fdm sa fet\u00e1lna frakcia &lt; 4 % vyskytovala u tehotn\u00fdch s hmotnos\u0165ou 60 kg len u 0,7 % \u017eien, pri hmotnosti 100 kg to u\u017e bolo u 7,1 % \u017eien a hmotnosti 160 kg a\u017e 51,1 % \u017eien(29). Tento \u00fadaj je d\u00f4le\u017eit\u00fd preto, \u017ee n\u00edzka fet\u00e1lna frakcia &lt; 4 % je aj v s\u00fa\u010dasnosti pou\u017e\u00edvan\u00fdm \u0161tandardom pre neinformat\u00edvnos\u0165 v\u00fdsledku NIPT pre detekciu triz\u00f3mi\u00ed chromoz\u00f3mov 21, \u00a018 \u00a0a 13 \u00a0v pr\u00edpade anal\u00fdzy \u00a0celogen\u00f3mov\u00fdm skenom cirkuluj\u00facej DNA tehotnej. V pr\u00edpade determin\u00e1cie cel\u00e9ho fet\u00e1lneho gen\u00f3mu z cirkuluj\u00facej \u00a0matern\u00e1lnej DNA sa aj v\u010faka spomenut\u00fdm poznatkom v jednej z najaktu\u00e1lnej\u0161\u00edch \u0161t\u00fadi\u00ed podarilo \u00a0pri gen\u00f3movom resekvenovan\u00ed s pokryt\u00edm 270x (haploidn\u00fd gen\u00f3m) zv\u00fd\u0161i\u0165 senzitivitu detekcie <em>de novo <\/em>patog\u00e9nnych variantov \u00a0a\u017e na 85 % a zv\u00fd\u0161enie pozit\u00edvnej\u00a0 predikt\u00edvnej hodnoty na 74 % (169-n\u00e1sobn\u00e9 zlep\u0161enie v porovnan\u00ed s predch\u00e1dzaj\u00facou \u0161t\u00fadiou).<\/p>\n<p>Uveden\u00e9 zlep\u0161enie senzitivity a pozit\u00edvnej predikt\u00edvnej hodnoty d\u00e1va v\u00fdznamn\u00fd predpoklad na vyu\u017e\u00edvanie\u00a0 NIPT nielen v r\u00e1mci detekcie chromoz\u00f3mov\u00fdch aber\u00e1ci\u00ed, ale dokonca aj pri neinvaz\u00edvnom prenat\u00e1lnom skr\u00edningu zameranom na monog\u00e9nne dedi\u010dn\u00e9 ochorenia, \u010d\u00edm sa vyu\u017eitie NIPT v\u00fdznamne roz\u0161\u00edri. Z\u00e1rove\u0148 \u00a0bola zisten\u00e1 nov\u00e1 skuto\u010dnos\u0165 \u010falej rozv\u00edjaj\u00faca poznatok o rozdielnej ve\u013ekostnej charakteristike fet\u00e1lnej a matern\u00e1lnej DNA. Bolo preuk\u00e1zan\u00e9, \u017ee rozdiel nie je len vo ve\u013ekosti t\u00fdchto \u00a0p\u00f4vodom odli\u0161n\u00fdch fragmentov, ale aj v lokaliz\u00e1cii \u00a0miest v gen\u00f3me, kde k \u0161tiepeniu genomickej DNA pri fragment\u00e1cii doch\u00e1dza <strong><em>(obr\u00e1zok 4)<\/em><\/strong>(30). To m\u00f4\u017ee \u00a0v\u00fdrazn\u00fdm sp\u00f4sobom pom\u00f4c\u0165 \u00a0k \u010fal\u0161iemu zv\u00fd\u0161eniu \u00a0\u0161pecificity NIPT, napr. pri presnej\u0161om v\u00fdpo\u010dte fet\u00e1lnej frakcie, ktor\u00e1 sl\u00fa\u017ei ako jeden zo z\u00e1kladn\u00fdch parametrov pri \u0161tatistick\u00fdch a bioinformatick\u00fdch anal\u00fdzach z\u00edskan\u00fdch d\u00e1t.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Cirkuluj\u00fac<\/strong><strong>a <\/strong><strong>n\u00e1do<\/strong><strong>r<\/strong><strong>ov<\/strong><strong>\u00e1 DNA \u2013 paraleln\u00e9 vyu\u017eitie<\/strong><\/p>\n<p><strong>v\u00fdsledk<\/strong><strong>o<\/strong><strong>v v\u00fdskumu <\/strong><strong>z cffDNA<\/strong><\/p>\n<p>Paralelne s v\u00fdskumom v oblasti anal\u00fdzy cffDNA bolo mo\u017en\u00e9 sledova\u0165 \u00a0podobn\u00fd v\u00fdskum \u00a0v oblasti anal\u00fdzy \u00a0cirkuluj\u00facej n\u00e1dorovej DNA (ctDNA \u2013 circulating tumor \u00a0DNA). Z\u00e1kladn\u00e9 poznatky o pr\u00edtomnosti a mno\u017estve ctDNA u pacientov s r\u00f4znymi n\u00e1dorov\u00fdmi ochoreniami boli detailne \u0161tudovan\u00e9, av\u0161ak do prechodu na \u00a0genomick\u00e9 met\u00f3dy anal\u00fdzy \u00a0univerz\u00e1lnymi met\u00f3dami, ktor\u00e9 \u00a0neumo\u017e\u0148ovali \u0161pecifick\u00fa detekciu, a teda ani kvantifik\u00e1ciu ctDNA, bola ich aplik\u00e1cia v klinickej praxi relat\u00edvne obmedzen\u00e1. Po zaveden\u00ed met\u00f3d MPS sa v\u0161ak postupne preuk\u00e1zalo, \u017ee ctDNA mo\u017eno \u0161pecificky detegova\u0165, a to kvalitat\u00edvne (detekcia n\u00e1dorovo \u0161pecifick\u00fdch patog\u00e9nnych zmien v cirkuluj\u00facej DNA) aj kvantitat\u00edvne (mno\u017estvo ctDNA koreluje s ve\u013ekos\u0165ou n\u00e1dorovej masy) \u00a0a tie\u017e s vy\u0161\u0161ou senzitivitou a \u0161pecificitou ako pri vyu\u017eit\u00ed cirkuluj\u00facich n\u00e1dorov\u00fdch buniek(31). Len ned\u00e1vno bola publikovan\u00e1 pr\u00e1ca, \u00a0ktor\u00e1 sledovala n\u00e1dorovo \u0161pecifick\u00e9 zmeny identifikovan\u00e9 v podobe glob\u00e1lnej hypometyl\u00e1cie v plazme pacienta s hepatocelul\u00e1rnym karcin\u00f3mom <strong><em>(obr\u00e1zok \u00a05)<\/em><\/strong>.<\/p>\n<p>Genomick\u00e1 anal\u00fdza zameran\u00e1 na hypometyl\u00e1ciu a anal\u00fdzu \u00a0CNV pouk\u00e1zala na r\u00fdchle odb\u00faranie ctDNA po odstr\u00e1nen\u00ed n\u00e1dorov\u00e9ho tkaniva \u00a0a tie\u017e na mo\u017enos\u0165 vyu\u017eitia tejto univerz\u00e1lnej na z\u00e1rove\u0148 n\u00e1dorovo \u0161pecifickej anal\u00fdzy zameranej na sledovanie pr\u00edtomnosti a mno\u017estva ctDNA, na monitoring, ke\u010f bolo mo\u017en\u00e9 vidie\u0165 udr\u017eanie \u010di n\u00e1rast \u00a0t\u00fdchto \u00a0n\u00e1dorovo \u0161pecifick\u00fdch zmien \u00a0v cirkuluj\u00facej \u00a0DNA u pacientov s ne\u00faspe\u0161n\u00fdm chirurgick\u00fdm \u00a0odstr\u00e1nen\u00edm n\u00e1dorov\u00e9ho tkaniva \u010di relapsom ochorenia(32). V s\u00fa\u010dasnosti mo\u017eno u\u017e aj v klinickej rutinnej \u00a0praxi vyu\u017ei\u0165 molekul\u00e1rne testy \u00a0zameran\u00e9 na neinvaz\u00edvnu detekciu n\u00e1dorovo \u0161pecifick\u00fdch genetick\u00fdch zmien. \u00a0Navy\u0161e \u00a0mo\u017eno pri tak\u00fdchto anal\u00fdzach vyu\u017ei\u0165 na MPS alternat\u00edvne, pre rutinn\u00fa laborat\u00f3rnu prax efekt\u00edvnej\u0161ie a jednoduch\u0161ie vysokopriepustn\u00e9 met\u00f3dy ich identifik\u00e1cie, ako napr. digit\u00e1lnu PCR alebo tzv. Beaming\u00a0 met\u00f3du, ktor\u00e9 s\u00fa schopn\u00e9 identifikova\u0165 n\u00e1dorovo \u0161pecifick\u00e9 bodov\u00e9 mut\u00e1cie a kvantifikova\u0165 ich s vysokou \u00a0presnos\u0165ou. Ako v s\u00fa\u010dasnosti komer\u010dne dostupn\u00e9 analyza\u010dn\u00e9 komplety mo\u017eno spomen\u00fa\u0165 komplety na identifik\u00e1ciu d\u00f4le\u017eit\u00fdch patog\u00e9nnych \u00a0a farmakogeneticky relevantn\u00fdch variantov v g\u00e9noch <em>KRAS<\/em>, <em>NRAS <\/em>a <em>EGFR <\/em>relevantn\u00fdch pre\u00a0 pacientov s kolorekt\u00e1lnym a nemalobunkov\u00fdm p\u013e\u00facnym karcin\u00f3mom. Podobne ako pri cffDNA bolo aj pri ctDNA preuk\u00e1zan\u00e9, \u017ee t\u00e1to \u00a0je viac fragmentovan\u00e1 ako zvy\u0161n\u00e1 cirkuluj\u00faca DNA, pri\u010dom ctDNA je priemerne najviac zast\u00fapen\u00e1 vo ve\u013ekosti 144 bp, k\u00fdm nen\u00e1dorov\u00e1 cirkuluj\u00faca DNA vo ve\u013ekosti 165 bp(33). Op\u00e4\u0165 je mo\u017en\u00e9, \u017ee ve\u013ekostn\u00e1 selekcia zameran\u00e1 na obohatenie ctDNA ve\u013ekostnej frakcie\u00a0 m\u00f4\u017ee \u00a0vies\u0165 k zv\u00fd\u0161eniu \u00a0senzitivity\u00a0 \u010di \u0161pecificity pou\u017eit\u00fdch molekul\u00e1rnych met\u00f3d anal\u00fdzy DNA, ako to bolo pri NIPT. Aktu\u00e1lne najnov\u0161\u00edm a vysokoperspekt\u00edvnym v\u00fdsledkom v\u00fdskumu cirkuluj\u00facich DNA je mo\u017enos\u0165 identifik\u00e1cie tkaniva, z ktor\u00e9ho v cirkuluj\u00facej \u00a0DNA sa nach\u00e1dzaj\u00face DNA molekuly \u00a0poch\u00e1dzaj\u00fa, tzv. \u00a0tkanivov\u00e9 \u00a0mapovanie. Vyu\u017eitie mo\u017enosti \u00a0takejto identifik\u00e1cie je \u00a0aplikovate\u013en\u00e9 v r\u00f4znych klinick\u00fdch \u0161pecializ\u00e1ci\u00e1ch nielen \u00a0v NIPT \u010di n\u00e1dorovej diagnostike a prognostike <strong><em>(obr\u00e1zok \u00a06)<\/em><\/strong>(34). Podobne ho mo\u017eno vyu\u017ei\u0165 tam, kde je potrebn\u00e9 sledova\u0165 \u00a0n\u00e1rast po\u0161kodenia tkan\u00edv prejavuj\u00face sa odumieran\u00edm buniek \u00a0(kardiovaskul\u00e1rne zlyhanie, posttransplanta\u010dn\u00fd \u00a0monitoring, extr\u00e9mna n\u00e1maha, a pod.)(34,35).<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Z\u00e1ver<\/strong><\/p>\n<p>Anal\u00fdza cirkuluj\u00facich \u00a0nukleov\u00fdch \u00a0kysel\u00edn\u00a0 sa v s\u00fa\u010dasnosti ukazuje ako najvhodnej\u0161\u00ed sp\u00f4sob neinvaz\u00edvneho DNA skr\u00edningu, prognostiky a diagnostiky s mnoh\u00fdmi klinick\u00fdmi aplik\u00e1ciami. V\u00fdskum za posledn\u00fdch p\u00e1r rokov uk\u00e1zal, \u00a0\u017ee ju\u00a0mo\u017eno zu\u017eitkova\u0165 \u00a0pri prenat\u00e1lnom genetickom \u00a0skr\u00edningu, v r\u00e1mci \u00a0ktor\u00e9ho bola \u00a0najm\u00e4 zo\u00a0 za\u010diatku intenz\u00edvne \u0161tudo- van\u00e1, \u00a0ale po celosvetovom roz\u0161\u00edren\u00ed genomick\u00fdch analyz\u00e1torov aj pri rutinn\u00fdch \u00a0anal\u00fdzach cirkuluj\u00facej \u00a0n\u00e1dorovej DNA v oblasti diagnostiky kardiovaskul\u00e1rneho po\u0161kodenia \u010di po- sttransplanta\u010dn\u00e9ho monitoringu, zalo\u017eenej na\u00a0 univerz\u00e1lnej met\u00f3de tkanivov\u00e9ho mapovania.<\/p>\n<p>&nbsp;<\/p>\n<p><strong><em>P<\/em><\/strong><strong><em>o\u010fako<\/em><\/strong><strong><em>v<\/em><\/strong><strong><em>anie<\/em><\/strong><\/p>\n<p><em>Publik\u00e1cia je v\u00fdsledkom implement\u00e1cie projektu podpore<\/em><em>n\u00e9ho \u00a0grantom \u00a0APVV-14-0273 a APVV-15-0327 Agent\u00fary \u00a0pre podporu v\u00fdskumu a v\u00fdvoja.<\/em><\/p>\n<p>&nbsp;<\/p>\n<p>LITERAT\u00faRA<br \/>\n1. Mandel P, Metais P. Les acides nucl\u00e9iques du plasma sanguin chez<br \/>\nl\u2019homme. C R Seances Soc Biol Fil 1948; 142(3-4): 241-3.<br \/>\n2. Tan EM, Schur PH, Carr RI, Kunkel HG. Deoxybonucleic acid (DNA) and<br \/>\nantibodies to DNA in the serum of patients with systemic lupus erythematosus.<br \/>\nJ Clin Invest 1966; 45(11): 1732-40.<br \/>\n3. Koffler D, Agnello V, Winchester R, Kunkel HG. The occurrence of single-<br \/>\nstranded DNA in the serum of patients with systemic lupus erythematosus<br \/>\nand other diseases. J Clin Invest 1973; 52(1): 198-204.<br \/>\n4. Leon SA, Shapiro B, Sklaroff DM, Yaros MJ. Free DNA in the serum of<br \/>\ncancer patients and the effect of therapy. Cancer Res 1977; 37(3): 646-50.<br \/>\n5. Lo YM, Patel P, Wainscoat JS, et al. Prenatal sex determination by<br \/>\nDNA amplification from maternal peripheral blood. Lancet 1989; 2(8676):<br \/>\n1363-5.<br \/>\n6. Lo YM, Tein MS, Lau TK, et al. Quantitative analysis of fetal DNA in<br \/>\nmaternal plasma and serum: implications for noninvasive prenatal diagnosis.<br \/>\nAm J Hum Genet 1998; 62(4): 768-75.<br \/>\n7. Lo YM, Zhang J, Leung TN, et al. Rapid clearance of fetal DNA from maternal<br \/>\nplasma. Am J Hum Genet 1999; 64(1): 218-24.<br \/>\n8. Krabchi K, Gros-Louis F, Yan J, et al. Quantification of all fetal nucleated<br \/>\ncells in maternal blood between the 18th and 22nd weeks of pregnancy<br \/>\nusing molecular cytogenetic techniques. Clin Genet 2001; 60(2): 145-50.<br \/>\n9. Mergenthaler S, Babochkina T, Kiefer V, et al. FISH analysis of all fetal<br \/>\nnucleated cells in maternal whole blood: improved specificity by the<br \/>\nuse of two Y-chromosome probes. J Histochem Cytochem 2005; 53(3):<br \/>\n319-22.<br \/>\n10. Bianchi DW, Zickwolf GK, Weil GJ, et al. Male fetal progenitor cells persist<br \/>\nin maternal blood for as long as 27 years postpartum. Proc Natl Acad<br \/>\nSci U S A 1996; 93(2): 705-8.<br \/>\n11. Lo YM, Hjelm NM, Fidler C, Set al. Prenatal diagnosis of fetal RhD<br \/>\nstatus by molecular analysis of maternal plasma. N Engl J Med 1998;<br \/>\n339(24): 1734-8.<br \/>\n12. Saito H, Sekizawa A, Morimoto T, et al. Prenatal DNA diagnosis of<br \/>\na single-gene disorder from maternal plasma. Lancet 2000; 356(9236):<br \/>\n1170.<br \/>\n13. Amicucci P, Gennarelli M, Novelli G, Dallapiccola B. Prenatal diagnosis<br \/>\nof myotonic dystrophy using fetal DNA obtained from maternal plasma.<br \/>\nClin Chem 2000; 46(2): 301-2.<br \/>\n14. Chan KC, Zhang J, Hui AB, et al. Size distributions of maternal and fetal<br \/>\nDNA in maternal plasma. Clin Chem 2004; 50(1): 88-92.<br \/>\n15. Li Y, Holzgreve W, Page-Christiaens GC, et al. Improved prenatal detection<br \/>\nof a fetal point mutation for achondroplasia by the use of size-fractionated<br \/>\ncirculatory DNA in maternal plasma \u2013 case report. Prenat Diagn<br \/>\n2004; 24(11): 896-8.<br \/>\n16. Li Y, Di Naro E, Vitucci A, et al. Detection of paternally inherited fetal<br \/>\npoint mutations for beta-thalassemia using size-fractionated cell-free<br \/>\nDNA in maternalplasma. JAMA. 2005 Feb 16;293(7):843-9. Erratum in:<br \/>\nJAMA 2005; 293(14):1728.<br \/>\n17. Li Y, Zimmermann B, Rusterholz C, et al. Size separation of circulatory<br \/>\nDNA in maternal plasma permits ready detection of fetal DNA polymorphisms.<br \/>\nClin Chem 2004; 50(6): 1002-11.<br \/>\n18. Chan KC, Ding C, Gerovassili A, et al. Hypermethylated RASSF1A in<br \/>\nmaternal plasma: A universal fetal DNA marker that improves the reliability<br \/>\nof noninvasive prenatal diagnosis. Clin Chem 2006; 52(12): 2211-8.<br \/>\n19. Lo YM, Lun FM, Chan KC, et al. Digital PCR for the molecular detection<br \/>\nof fetal chromosomal aneuploidy. Proc Natl Acad Sci U S A 2007; 104(32):<br \/>\n13116-21.<br \/>\nPreh\u013eadov\u00e9 pr\u00e1ce<br \/>\n52<br \/>\n1\/2017<br \/>\n20. Fan HC, Blumenfeld YJ, Chitkara U, et al. Noninvasive diagnosis of<br \/>\nfetal aneuploidy by shotgun sequencing DNA from maternal blood. Proc<br \/>\nNatl Acad Sci U S A 2008; 105(42): 16266-71.<br \/>\n21. Chiu RW, Chan KC, Gao Y, et al. Noninvasive prenatal diagnosis of fetal<br \/>\nchromosomalaneuploidy by massively parallel genomic sequencing ofDNA<br \/>\nin maternal plasma. Proc Natl Acad Sci U S A 2008; 105(51): 20458-<br \/>\n63.<br \/>\n22. Lo YM, Chan KC, Sun H, et al. Maternal plasma DNA sequencing reveals<br \/>\nthe genome-wide genetic and mutational profile of the fetus. Sci<br \/>\nTransl Med 2010; 2(61): 61ra91.<br \/>\n23. Lun FM, Tsui NB, Chan KC, et al. Noninvasive prenatal diagnosis of<br \/>\nmonogenic diseases by digital size selection and relative mutation dosage<br \/>\non DNA in maternal plasma. Proc Natl Acad Sci U S A 200; 105(50):<br \/>\n19920-5.<br \/>\n24. Chiu RW, Akolekar R, Zheng YW, et al. Non-invasive prenatal assessment<br \/>\nof trisomy 21 by multiplexed maternal plasma DNA sequencing:<br \/>\nlarge scale validity study. BMJ 2011; 342: c7401.<br \/>\n25. Chen EZ, Chiu RW, Sun H, et al. Noninvasive prenatal diagnosis of fetal<br \/>\ntrisomy 18 and trisomy 13 by maternal plasma DNA sequencing. PLoS<br \/>\nOne 2011; 6(7): e21791.<br \/>\n26. Ashoor G, Syngelaki A, Wagner M, et al. Chromosome-selective sequencing<br \/>\nof maternal plasma cell-free DNA for first-trimester detection of<br \/>\ntrisomy 21 and trisomy 18. Am J Obstet Gynecol 2012; 206(4): 322.e1-5.<br \/>\n27. Bianchi DW, Platt LD, Goldberg JD, et al; MatErnal BLood IS Source<br \/>\nto Accurately diagnose fetal aneuploidy (MELISSA) Study Group. Genome-<br \/>\nwide fetal aneuploidy detection by maternal plasma DNA sequencing.<br \/>\nObstet Gynecol 2012; 119(5): 890-901.<br \/>\n28. Wang E, Batey A, Struble C, et al. Gestational age and maternal weight<br \/>\neffects on fetal cell-free DNA in maternal plasma. Prenat Diagn 2013;<br \/>\n33(7): 662-6.<br \/>\n29. Ashoor G, Syngelaki A, Poon LC, et al. Fetal fraction in maternal plasma<br \/>\ncell-free DNA at 11-13 weeks\u2018 gestation: relation to maternal and fetal<br \/>\ncharacteristics. Ultrasound Obstet Gynecol 2013; 41(1): 26-32.<br \/>\n30. Chan KC, Jiang P, Sun K, et al. Second generation noninvasive fetal<br \/>\ngenome analysis reveals de novo mutations, single-base parental inheritance,<br \/>\nand preferred DNA ends. Proc Natl Acad Sci U S A 2016; 113(50):<br \/>\nE8159-E8168.<br \/>\n31. Punnoose EA, Atwal S, Liu W, et al. Evaluation of circulating tumor<br \/>\ncells and circulating tumor DNA in non-small cell lung cancer: association<br \/>\nwith clinical endpoints in a phase II clinical trial of pertuzumab and<br \/>\nerlotinib. Clin Cancer Res 2012; 18(8): 2391-401.<br \/>\n32. Chan KC, Jiang P, Chan CW, et al. Noninvasive detection of cancer-associated<br \/>\ngenome-wide hypomethylation and copy number aberrations by<br \/>\nplasma DNA bisulfite sequencing. Proc Natl Acad Sci U S A 2013; 110(47):<br \/>\n18761-8.<br \/>\n33. Underhill HR, Kitzman JO, Hellwig S, et al. Fragment Length of Circulating<br \/>\nTumor DNA. PLoS Genet 2016; 12(7): e1006162.<br \/>\n34. Sun K, Jiang P, Chan KC, et al. Plasma DNA tissue mapping by genome-<br \/>\nwide methylationsequencing for noninvasive prenatal, cancer, and<br \/>\ntransplantation assessments. Proc Natl Acad Sci U S A 2015; 112(40):<br \/>\nE5503-12.<br \/>\n35. Atamaniuk J, Vidotto C, Tschan H, et al. Increased concentrations of<br \/>\ncell-free plasma DNA after exhaustive exercise. Clin Chem 2004; 50(9):<br \/>\n1668-70.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>*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. &nbsp; \u00davod D\u00f4le\u017eitos\u0165 objavenia cirkuluj\u00facej DNA v roku 1947 Mandelom a Metaisom bola \u00a0dlh\u00fd \u010das \u00a0nerozpoznan\u00e1(1). A\u017e o dvadsa\u0165 rokov, v roku 1966, \u00a0bola \u00a0publikovan\u00e1 \u0161t\u00fadia poukazuj\u00faca na asoci\u00e1ciu medzi<\/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":[608,610,611,612,609],"class_list":["post-1011","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-circulating-dna","tag-circulating-free-fetal-dna","tag-circulating-tumor-dna","tag-massive-parallel-sequencing","tag-non-invasive-dna-testing","typ_clanku-review-article"],"acf":{"abstrakt":"<p>Analysis of circulating nucleic acids has acquired a strong position in biomedical research. This position is linked to discoveries related to transformation of analyses from genetical to genomic, an extensive development in bioinformatics, technological advance and availability of genomic analysers. Besides other benefits, robustness of the currently used genomic methods and their capacity to deliver previously unprecedented results of both qualitative and quantitative character enabled the development of new possibilities in research of the so called non-invasive testing.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Key words:<\/strong> circulating DNA, non-invasive DNA testing, circulating free fetal DNA, circulating tumor DNA, massive parallel sequencing<\/p>\n","casopis":[{"ID":735,"post_author":"7","post_date":"2017-04-06 13:21:01","post_date_gmt":"2017-04-06 11:21:01","post_content":"<ul>\r\n \t<li>Pseudoglandular nevus \u2013 a rare morphology of melanocytic nevus (case report)<\/li>\r\n \t<li>Differential molecular diagnosis of multiple myeloma and Waldenstr\u00f6m macroglobulinemia<\/li>\r\n \t<li>Molecular analysis of prognostically significant markers of chronic lymphocytic leukemia<\/li>\r\n \t<li>Prevalence of <em>Streptococcus pneumoniae<\/em> phyla in inflammatory diseases of upper airways in preschool age children and their resistance to antibiotics<\/li>\r\n \t<li>Malign melanoma - new aspects of research<\/li>\r\n \t<li style=\"list-style-type: none;\"><\/li>\r\n<\/ul>","post_title":"Newslab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-2017-1","to_ping":"","pinged":"","post_modified":"2017-08-16 21:11:52","post_modified_gmt":"2017-08-16 19:11:52","post_content_filtered":"","post_parent":0,"guid":"http:\/\/www.newslab.sk\/?post_type=casopis&#038;p=735\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"48","upload_clanok":{"ID":1012,"id":1012,"title":"Cirkuluj\u00faca DNA \u2013 od jej objavenia ....","filename":"Cirkuluj\u00faca-DNA-\u2013-od-jej-objavenia-.....pdf","filesize":830655,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2017\/04\/Cirkuluj\u00faca-DNA-\u2013-od-jej-objavenia-.....pdf","link":"https:\/\/www.newslab.sk\/en\/circulating-dna-from-its-discovery-to-non-invasive-prenatal-testing-of-aneuploidies-and-monogenic-hereditary-diseases-to-non-invasive-cancer-diagnosis-and-prognostics\/cirkulujuca-dna-od-jej-objavenia\/","alt":"","author":"7","description":"","caption":"","name":"cirkulujuca-dna-od-jej-objavenia","status":"inherit","uploaded_to":1011,"date":"2017-04-06 12:02:39","modified":"2017-04-06 12:02:39","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\/1011","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=1011"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1011\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1011"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1011"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1011"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}