{"id":1588,"date":"2018-11-11T19:14:04","date_gmt":"2018-11-11T18:14:04","guid":{"rendered":"http:\/\/www.newslab.sk\/2018\/11\/11\/cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-pacientov-s-nadorovymi-ochoreniami\/"},"modified":"2018-11-11T19:18:38","modified_gmt":"2018-11-11T18:18:38","slug":"cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-pacientov-s-nadorovymi-ochoreniami","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-pacientov-s-nadorovymi-ochoreniami\/","title":{"rendered":"Circulating tumour cells as markers usable in management of 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

 <\/p>\n

Cirkuluj\u00face n\u00e1dorov\u00e9 bunky<\/strong><\/p>\n

Cirkuluj\u00face n\u00e1dorov\u00e9 bunky (CTC) s\u00fa bunky uvo\u013e\u0148ovan\u00e9 do krvn\u00e9ho rie\u010diska z prim\u00e1rneho n\u00e1doru u\u017e v\u010dasne po\u010das jeho formovania a rastu a\/alebo jeho metast\u00e1zami. S\u00fa preuk\u00e1zate\u013ene asociovan\u00e9 s metast\u00e1zovan\u00edm, pri\u010dom s\u00fa schopn\u00e9 inv\u00e1zie do okolia cez extracelul\u00e1rnu matrix a vrstvy strom\u00e1lnych buniek, nasleduje intravaz\u00e1cia t\u00fdchto buniek do l\u00famenu ciev, ich transport krvn\u00fdm rie\u010diskom, zachytenie v inej vzdialenej lokalite a n\u00e1sledn\u00e1 extravaz\u00e1cia do parench\u00fdmu vzdialen\u00e9ho tkaniva. Bunky \u010falej pre\u017e\u00edvaj\u00fa v novom mikroprostred\u00ed a maj\u00fa potenci\u00e1l tvorby mikrometast\u00e1z, ak znovu spustia svoj proliferat\u00edvny program na danom mieste, \u010d\u00edm d\u00e1vaj\u00fa vznik makroskopick\u00fdm, klinicky detegovate\u013en\u00fdm neoplastick\u00fdm \u00fatvarom(1).<\/p>\n

Z cytologick\u00e9ho poh\u013eadu s\u00fa CTCs heterog\u00e9nnou skupinou buniek obsahuj\u00facou \u017eivotaschopn\u00e9, apoptick\u00e9, dormantn\u00e9 (spiace) bunky, ako aj bunky schopn\u00e9 zalo\u017ei\u0165 metast\u00e1zu. Ich vn\u00fatrobunkov\u00e9 zmeny m\u00f4\u017eu vies\u0165 k vzniku diseminovan\u00fdch n\u00e1dorov\u00fdch buniek (DTCs), ktor\u00e9 s\u00fa pova\u017eovan\u00e9 za samotn\u00e9 mikrometast\u00e1zy. K\u00fdm CTCs s\u00fa n\u00e1dorov\u00e9 bunky pr\u00edtomn\u00e9 v krvnom obehu onkologick\u00fdch pacientov, DTCs s\u00fa n\u00e1dorov\u00e9 bunky, ktor\u00e9 sa u\u017e us\u00eddlili v cie\u013eovom tkanive, napr\u00edklad v kostnej dreni. DTCs s\u00fa naj\u010dastej\u0161ie detegovan\u00e9 a op\u00edsan\u00e9 pr\u00e1ve v kostnej dreni onkologick\u00fdch pacientov. Jedn\u00fdm z d\u00f4vodov je aj to, \u017ee tento org\u00e1n je \u013eah\u0161ie pr\u00edstupn\u00fd pre biopsiu<\/p>\n

v porovnan\u00ed s in\u00fdmi org\u00e1nmi, ako je pe\u010de\u0148, p\u013e\u00faca alebo mozog. DTCs m\u00f4\u017eu zosta\u0165 v spiacom stave aj nieko\u013eko rokov po odstr\u00e1nen\u00ed prim\u00e1rneho n\u00e1doru a n\u00e1sledne vyvola\u0165 vznik metast\u00e1z(2). Navy\u0161e, DTCs z t\u00fdchto metast\u00e1z sa m\u00f4\u017eu op\u00e4tovne uvo\u013eni\u0165 do krvn\u00e9ho obehu a \u0161\u00edri\u0165 sa jeho prostredn\u00edctvom do \u010fal\u0161\u00edch vzdialen\u00fdch tkan\u00edv a vytv\u00e1ra\u0165 tak sekund\u00e1rne metast\u00e1zy. Dokonca tak\u00e9to DTCs konvertovan\u00e9 sp\u00e4tne na CTCs m\u00f4\u017eu kolonizova\u0165 prim\u00e1rny n\u00e1dor a u\u017e existuj\u00face metast\u00e1zy v procese naz\u00fdvanom samov\u00fdsev (self-seeding), \u010d\u00edm d\u00e1vaj\u00fa mo\u017enos\u0165 vzniku agres\u00edvnych typov metast\u00e1z(3).<\/p>\n

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Koncentr\u00e1cia CTCs<\/strong><\/p>\n

Predpoklad\u00e1 sa, \u017ee jeden gram n\u00e1dorovej masy uvo\u013e\u0148uje denne desa\u0165tis\u00edce CTCs do krvn\u00e9ho obehu(4). Ich \u017eivotnos\u0165 v cirkul\u00e1cii je v\u0161ak \u010dasovo ve\u013emi obmedzen\u00e1 a pohybuje sa v rozmedz\u00ed 1 \u2013 2,4 hodiny(5). Odhaduje sa, \u017ee len asi 1 z 1 000 buniek zost\u00e1va v cirkul\u00e1cii \u017eivotaschopn\u00fdch a len 1 z 10 000 z t\u00fdchto CTCs je schopn\u00fdch vytvori\u0165 metast\u00e1zy(6). Je to tak pre nutnos\u0165 prekonania vplyvu hemodynamick\u00fdch s\u00edl a p\u00f4sobenia buniek imunitn\u00e9ho syst\u00e9mu (predov\u0161etk\u00fdm NK buniek)(4). Navy\u0161e k tomu prispieva strata kontaktu s okolit\u00fdmi bunkami, a t\u00fdm sp\u00f4soben\u00e1 smr\u0165 v procese anoik\u00f3zy. Vo v\u00fdsledku s\u00fa preto CTCs v perif\u00e9rnej krvi pr\u00edtomn\u00e9 vo ve\u013emi n\u00edzkej koncentr\u00e1cii, ktor\u00e1 sa pohybuje medzi 1 \u2013 10 bunkami na 10 ml krvi u v\u00e4\u010d\u0161iny onkologick\u00fdch pacientov(7).<\/p>\n

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Prechod CTCs do cirkul\u00e1cie a epiteli\u00e1lnomezenchym\u00e1lny prechod<\/strong><\/h2>\n

CTCs sa m\u00f4\u017eu do cirkul\u00e1cie dosta\u0165 dvomi sp\u00f4sobmi, a to akt\u00edvne \u2013 vycestovan\u00edm buniek so zv\u00fd\u0161en\u00fdm migra\u010dn\u00fdm potenci\u00e1lom, alebo pas\u00edvne \u2013 uvo\u013e\u0148ovan\u00edm bu\u010f jednotliv\u00fdch buniek, pr\u00edpadne bunkov\u00fdch zhlukov. Biologicky v\u00fdznamnej\u0161ie s\u00fa pr\u00e1ve tie CTCs, ktor\u00e9 s\u00fa schopn\u00e9 akt\u00edvne vycestova\u0165, \u010do je spojen\u00e9 s ich vstupom do procesu epiteli\u00e1lno-mezenchym\u00e1lneho prechodu (EMT)(8).<\/p>\n

Epiteli\u00e1lno-mezenchym\u00e1lny prechod je proces, ktor\u00fd sa fyziologicky uplat\u0148uje po\u010das embryogen\u00e9zy a zah\u0155\u0148a migr\u00e1ciu ektoderm\u00e1lnych buniek v embryu po\u010das gastrul\u00e1cie za vzniku mezodermu (EMT typu I)(9). V neskor\u0161om obdob\u00ed hr\u00e1 ve\u013emi d\u00f4le\u017eit\u00fa \u00falohu po\u010das hojenia r\u00e1n, regener\u00e1cie tkan\u00edv a fibr\u00f3zy org\u00e1nov (EMT typu II)(10) a z poh\u013eadu n\u00e1dorov\u00e9ho tkaniva je spojen\u00fd s metastatick\u00fdm \u0161\u00edren\u00edm (EMT typu III). Umo\u017e\u0148uje diferencovan\u00fdm epitelov\u00fdm bunk\u00e1m, ktor\u00e9 interaguj\u00fa s baz\u00e1lnou membr\u00e1nou prostredn\u00edctvom ich povrchu, podst\u00fapi\u0165 tak\u00e9 biologick\u00e9 zmeny, ktor\u00e9 im umo\u017e\u0148uj\u00fa z\u00edska\u0165 fenotyp mezenchym\u00e1lnych buniek. Ten zah\u0155\u0148a zv\u00fd\u0161en\u00fa migra\u010dn\u00fa kapacitu, invaz\u00edvnos\u0165, zv\u00fd\u0161en\u00fa odolnos\u0165 proti apopt\u00f3ze a zna\u010dne zv\u00fd\u0161en\u00fa produkciu zlo\u017eiek extracelul\u00e1rnej matrix (ECM)(8). Dokon\u010denie EMT je signalizovan\u00e9 degrad\u00e1ciou baz\u00e1lnej membr\u00e1ny a vytvorenie mezenchym\u00e1lnych buniek, ktor\u00e9 s\u00fa schopn\u00e9 migrova\u0165 z epitelovej vrstvy, z ktorej vznikli (obr\u00e1zok 1)<\/em><\/strong>.<\/p>\n

EMT je zlo\u017eit\u00fd proces, do ktor\u00e9ho je zapojen\u00fdch viacero sign\u00e1lnych dr\u00e1h, preto\u017ee bunky musia prejs\u0165 funk\u010dn\u00fdmi aj morfologick\u00fdmi zmenami. EMT m\u00f4\u017ee by\u0165 indukovan\u00fd r\u00f4znymi mechanizmami, medzi ktor\u00e9 patr\u00ed z\u00e1pal, acid\u00f3za, hypoxia, rastov\u00e9 faktory a v neposlednom rade aj vplyv mikroprostredia n\u00e1doru, ktor\u00e9 m\u00f4\u017ee stimulova\u0165 bunkov\u00fa migr\u00e1ciu a inv\u00e1ziu. Na druhej strane v\u0161ak ide o reverzibiln\u00fd proces, ke\u010f cirkuluj\u00face n\u00e1dorov\u00e9 bunky po dosiahnut\u00ed vzdialen\u00e9ho org\u00e1nu a extravaz\u00e1cii do jeho parench\u00fdmu z\u00edskavaj\u00fa znovu fenotyp epiteli\u00e1lnych buniek v procese naz\u00fdvanom mezenchym\u00e1lno-epiteli\u00e1lny prechod (MET), aby mohli \u010falej proliferova\u0165 ako epiteli\u00e1lne metastatick\u00e9 depozity.<\/p>\n

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N\u00e1dorov\u00e9 kme\u0148ov\u00e9 bunky<\/h2>\n

Mal\u00e1 \u010das\u0165 n\u00e1dorov\u00fdch buniek vykazuje vlastnosti dospel\u00fdch kme\u0148ov\u00fdch buniek, \u010di\u017ee maj\u00fa schopnos\u0165 samoobnovenia, s\u00fa multipotentn\u00e9 a d\u00e1vaj\u00fa tak mo\u017enos\u0165 vzniku diferencovan\u00fdch n\u00e1dorov\u00fdch buniek a s\u00fa schopn\u00e9 iniciova\u0165 rast n\u00e1doru. Tieto bunky s\u00fa ozna\u010dovan\u00e9 ako n\u00e1dorov\u00e9 kme\u0148ov\u00e9 bunky (CSCs), bunky podobn\u00e9 kme\u0148ov\u00fdm bunk\u00e1m, pr\u00edpadne ako bunky iniciuj\u00face n\u00e1dor a s\u00fa sp\u00e1jan\u00e9 s mezenchym\u00e1lnym fenotypom.<\/p>\n

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Cirkuluj\u00face n\u00e1dorov\u00e9 mikroemb\u00f3lie<\/h2>\n

Cirkuluj\u00face n\u00e1dorov\u00e9 bunky m\u00f4\u017eu v obehu cirkulova\u0165 ako samostatn\u00e9 bunky alebo z prim\u00e1rneho tumoru sa m\u00f4\u017ee odl\u00fa\u010di\u0165 z\u00e1rove\u0148 viacero buniek, ktor\u00e9 potom putuj\u00fa v zhlukoch. Tie m\u00f4\u017eu obsahova\u0165 od dvoch do viac ako 50 buniek a ozna\u010duj\u00fa sa ako cirkuluj\u00face n\u00e1dorov\u00e9 mikroemb\u00f3lie (CTMs). CTMs tvoria bu\u010f len n\u00e1dorov\u00e9 bunky, alebo m\u00f4\u017eu by\u0165 spojen\u00e9 s in\u00fdmi typmi buniek, ako s\u00fa fibroblasty, leukocyty, endoteli\u00e1lne bunky, pericyty, krvn\u00e9 do\u0161ti\u010dky(12). O v\u00fdznamnosti CTC\u00a0zhlukov sa dlho diskutovalo, av\u0161ak CTMs poch\u00e1dzaj\u00face od pacientov s malobunkov\u00fdm karcin\u00f3mom p\u013e\u00fac vykazovali nepr\u00edtomnos\u0165 apoptick\u00fdch buniek, \u010do im d\u00e1valo v\u00fdhodu v pre\u017e\u00edvan\u00ed, a tie\u017e n\u00edzky podiel proliferat\u00edvnych buniek, \u010d\u00edm sa st\u00e1vali rezistentnej\u0161\u00edmi proti chemoterapii(13). D\u00f4le\u017eit\u00fdm zisten\u00edm tie\u017e bolo, \u017ee bunky nach\u00e1dzaj\u00face sa v CTMs nevznikli delen\u00edm jedinej migruj\u00facej bunky, ale s\u00fa oligoklon\u00e1lneho p\u00f4vodu a hoci sa v cirkul\u00e1cii vyskytuj\u00fa ove\u013ea zriedkavej\u0161ie ako samostatn\u00e9 CTCs, maj\u00fa signifikantne vy\u0161\u0161\u00ed metastatick\u00fd potenci\u00e1l (23- a\u017e 50x) a ich pr\u00edtomnos\u0165 je spojen\u00e1 so zhor\u0161en\u00fdm celkov\u00fdm pre\u017e\u00edvan\u00edm(14).<\/p>\n

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Detekcia CTCs<\/h2>\n

Identifik\u00e1cia a charakteriz\u00e1cia CTCs si vy\u017eaduje vysokosenzit\u00edvne a \u0161pecifick\u00e9 met\u00f3dy, ke\u010f\u017ee CTCs sa v cirkul\u00e1cii nach\u00e1dzaj\u00fa vo ve\u013emi n\u00edzkej koncentr\u00e1cii, navy\u0161e s vysok\u00fdm pozad\u00edm in\u00fdch krvn\u00fdch buniek (na jednu CTC pripadaj\u00fa mili\u00f3ny in\u00fdch krvn\u00fdch buniek). Pri ich detekcii ide zvy\u010dajne o kombin\u00e1ciu izola\u010dn\u00fdch a detek\u010dn\u00fdch met\u00f3d. Na izol\u00e1ciu CTCs sa vyu\u017e\u00edvaj\u00fa r\u00f4zne technol\u00f3gie, ktor\u00e9 s\u00fa zalo\u017een\u00e9 na r\u00f4znych vlastnostiach CTCs, ktor\u00e9 ich odli\u0161uj\u00fa od norm\u00e1lnych hematopoetick\u00fdch buniek. Ide o vlastnosti fyzik\u00e1lneho (ve\u013ekos\u0165, denzita, elektrick\u00fd n\u00e1boj, tvarov\u00e1 flexibilita) aj biologick\u00e9ho charakteru (expresia prote\u00ednov na povrchu buniek, \u017eivotaschopnos\u0165).<\/p>\n

V\u00fdhodou technol\u00f3gi\u00ed vyu\u017e\u00edvaj\u00facich fyzik\u00e1lne vlastnosti je separ\u00e1cia CTCs bez ich farbenia. Patr\u00ed sem napr. centrifug\u00e1cia v hustotnom gradiente (Ficoll, OncoQuick), filtr\u00e1cia cez \u0161peci\u00e1lne filtre na z\u00e1klade ve\u013ekosti CTCs (ISET), v\u0161estrann\u00fd bio\u010dip vyu\u017e\u00edvaj\u00faci jedine\u010dn\u00e9 rozdiely vo ve\u013ekosti a deformovate\u013enosti n\u00e1dorov\u00fdch buniek (zachyt\u00e1van\u00e9 s\u00fa v\u00e4\u010d\u0161ie a tuh\u0161ie CTCs v porovnan\u00ed s krvn\u00fdmi bunkami), fotoakustick\u00e1 prietokov\u00e1 cytometria, dielektrofor\u00e9za (DEP)(15).<\/p>\n

Medzi technol\u00f3gie vyu\u017e\u00edvaj\u00face biologick\u00e9 vlastnosti CTCs patria imunofluorescen\u010dn\u00e9 met\u00f3dy. V tomto pr\u00edpade sa pou\u017e\u00edvaj\u00fa protil\u00e1tky proti n\u00e1dorovo asociovan\u00fdm antig\u00e9nom, ke\u010f ide o pozit\u00edvnu selekciu, alebo protil\u00e1tky proti be\u017en\u00e9mu leukocytov\u00e9mu antig\u00e9nu CD45, ke\u010f ide o negat\u00edvnu selekciu. Protil\u00e1tky proti dan\u00e9mu antig\u00e9nu s\u00fa naviazan\u00e9 na magnetick\u00fdch partikul\u00e1ch, ktor\u00e9 umo\u017e\u0148uj\u00fa separ\u00e1ciu CTCs naviazan\u00fdch na antig\u00e9n p\u00f4soben\u00edm magnetick\u00e9ho po\u013ea. Pri pozit\u00edvnej selekcii sa zvy\u010dajne vyu\u017e\u00edvaj\u00fa protil\u00e1tky proti adhez\u00edvnej molekule epitelov\u00fdch buniek (EpCAM). CTCs s\u00fa n\u00e1sledne detegovan\u00e9 imunocytologicky protil\u00e1tkami na cytokerat\u00edny (CKs). Medzi met\u00f3dy vyu\u017e\u00edvaj\u00face na separ\u00e1ciu CTCs EpCAM protil\u00e1tky patr\u00ed syst\u00e9m CellSearch\u00ae<\/strong>\u00a0\u00a0<\/strong>(Janssen Diagnostics, Raritan, NJ, USA). Tento syst\u00e9m umo\u017e\u0148uje stanovi\u0165 po\u010detnos\u0165 CTCs epiteli\u00e1lneho p\u00f4vodu pou\u017eit\u00edm magnetick\u00fdch partik\u00fal pokryt\u00fdch molekulami EpCAM a CTCs identifikuje pod\u013ea morfologick\u00fdch charakterist\u00edk, pozit\u00edvnej expresie cytokerat\u00ednov 8, 18 a\/alebo 19 a absencie hematopoetick\u00fdch antig\u00e9nov CD45(16). Ide o jedin\u00fd syst\u00e9m, ktor\u00fd schv\u00e1lil americk\u00fd \u00darad pre kontrolu liekov a potrav\u00edn (FDA) ako prostriedok na ur\u010denie progn\u00f3zy u pacientov s metastatickou rakovinou prsn\u00edka, prostaty a kolorekta(17-19).<\/p>\n

Nov\u00fdmi s\u013eubn\u00fdmi met\u00f3dami zalo\u017een\u00fdmi na EpCAM s\u00fa platformy vyu\u017e\u00edvaj\u00face mikrofluidick\u00fa technol\u00f3giu, najm\u00e4 CTC-\u010dip, Herringbone \u010dip, iCHIP, IsoFlux(12), ktor\u00e9 umo\u017e\u0148uj\u00fa sprac\u00fava\u0165 vzorky s ve\u013emi mal\u00fdm objemom.<\/p>\n

Biologick\u00fd v\u00fdznam CTCs epiteli\u00e1lneho p\u00f4vodu (EpCAM a bunky pozit\u00edvne na cytokerat\u00edn) je zrejm\u00fd z prognostickej d\u00f4le\u017eitosti CTCs, ktor\u00e9 boli zachyten\u00e9 CellSearch\u00ae <\/strong>syst\u00e9mom v r\u00f4znych typoch n\u00e1dorov(17-19). Ak v\u0161ak nejak\u00e1 \u010das\u0165 cirkuluj\u00facich n\u00e1dorov\u00fdch buniek podst\u00fapi EMT, ke\u010f s\u00fa downregulovan\u00e9 epiteli\u00e1lne markery, a, naopak, upregulovan\u00e9 tie mezenchym\u00e1lne, d\u00f4le\u017eit\u00e1 subpopul\u00e1cia CTCs nemus\u00ed by\u0165 met\u00f3dami, ktor\u00e9 s\u00fa z\u00e1visl\u00e9 od expresie EpCAM a cytokerat\u00ednov, zachyten\u00e1. Z tohto d\u00f4vodu sa na zachytenie CTCs neexprimuj\u00facich EpCAM vyu\u017e\u00edva kokteil protil\u00e1tok proti r\u00f4znym in\u00fdm epiteli\u00e1lnym povrchov\u00fdm antig\u00e9nom, napr. proti receptoru 2 \u013eudsk\u00e9ho epiderm\u00e1lneho rastov\u00e9ho faktora (HER2), receptoru pre epiderm\u00e1lny rastov\u00fd faktor (EGFR), muc\u00edn-1 (MUC1) a protil\u00e1tkam proti mezenchym\u00e1lnym a kme\u0148ov\u00fdm bunkov\u00fdm antig\u00e9nom (c-MET, N-kadher\u00edn, CD318 a antig\u00e9n mezenchym\u00e1lnych kme\u0148ov\u00fdch buniek)(20).<\/p>\n

Bez oh\u013eadu na v\u00fdber izola\u010dnej met\u00f3dy ani jednou jednokrokovou platformou nemo\u017eno dosiahnu\u0165 izol\u00e1ciu \u010dist\u00fdch CT- Cs. CTC frakcia oby\u010dajne st\u00e1le obsahuje aj zna\u010dn\u00fd po\u010det leukocytov. Z tohto d\u00f4vodu je potrebn\u00e1 n\u00e1sledn\u00e1 identifik\u00e1cia CTCs na \u00farovni buniek pou\u017eit\u00edm met\u00f3d umo\u017e\u0148uj\u00facich odl\u00ed\u0161i\u0165 n\u00e1dorov\u00e9 bunky od norm\u00e1lnych krvn\u00fdch buniek. Tieto met\u00f3dy s\u00fa zalo\u017een\u00e9 bu\u010f na detekcii prote\u00ednov, alebo expresii antig\u00e9nov asociovan\u00fdch tumor pomocou kvantitat\u00edvnej RT-PCR. V s\u00fa\u010dasnosti na detekciu viabiln\u00fdch CTCs z\u00edskan\u00fdch od onkologick\u00fdch pacientov existuj\u00fa dva rozdielne in vitro <\/em>testy. Prv\u00fdm je EPISPOT, ktor\u00fd deteguje \u0161pecifick\u00e9 prote\u00edny vylu\u010dovan\u00e9 CTCs do prostredia(21), druh\u00fdm je invaz\u00edvny test, ktor\u00fd sk\u00fama schopnos\u0165 CTCs tr\u00e1vi\u0165 fluorescen\u010dne zna\u010den\u00fa bunkov\u00fa matrix(7).<\/p>\n

Na detekciu viabiln\u00fdch CTCs mo\u017eno tie\u017e pou\u017ei\u0165 met\u00f3du kvantitat\u00edvnej RT-PCR, ke\u010f sa vyhodnocuje expresia \u0161pecifick\u00fdch mRNA molek\u00fal. V pr\u00edpade rakoviny prsn\u00edka sa naj\u010dastej\u0161ie vyu\u017e\u00edva mRNA k\u00f3duj\u00faca CK19. Nev\u00fdhodou v\u0161ak je, \u017ee t\u00e1to met\u00f3da neumo\u017e\u0148uje stanovenie po\u010detnosti buniek a navy\u0161e mnoho transkriptov (napr. k\u00f3duj\u00face CK18, CK19, CK20, MUC1, karcinoembryon\u00e1lny antig\u00e9n, prostatick\u00fd \u0161pecifick\u00fd antig\u00e9n) je v malom mno\u017estve exprimovan\u00fdch aj v norm\u00e1lnych krvn\u00fdch bunk\u00e1ch a bunk\u00e1ch kostnej drene(22), preto pre spr\u00e1vne vyhodnotenie a detekciu viabiln\u00fdch CTCs je potrebn\u00e1 valid\u00e1cia cut-off <\/em>hodnoty.<\/p>\n

 <\/p>\n

Klinick\u00e9 vyu\u017eitie cirkuluj\u00facich n\u00e1dorov\u00fdch buniek<\/h2>\n

Potenci\u00e1lne vyu\u017eitie CTCs v klinickej praxi je rozsiahle, od prognostick\u00e9ho a predikt\u00edvneho vyu\u017eitia a\u017e po detekciu minim\u00e1lnej rezidu\u00e1lnej choroby a monitorovania ochorenia (obr\u00e1zok 2)<\/em><\/strong>.<\/p>\n

CTCs ako prognostick\u00fd a farmakodynamick\u00fd biomarker. <\/strong>FDA schv\u00e1lila stanovenie po\u010detnosti CTCs s presn\u00fdmi cut-off <\/em>hodnotami a s vyu\u017eit\u00edm CellSearch\u00ae <\/strong>syst\u00e9mu ako prognostic- k\u00fd marker vybran\u00fdch n\u00e1dorov\u00fdch ochoren\u00ed. V pr\u00edpade rakovi- ny prsn\u00edka a prostaty je pr\u00edtomnos\u0165 piatich CTCs na 7,5 ml krvi spojen\u00e1 s krat\u0161\u00edm pre\u017e\u00edvan\u00edm pacientov, pri kolorekt\u00e1l- nom karcin\u00f3me (CRC) s\u00fa to 3 bunky na 7,5 ml krvi(17-19). Prognostick\u00e1 vyu\u017eite\u013enos\u0165 enumer\u00e1cie CTCs s vyu\u017eit\u00edm CellSearch\u00ae <\/strong>syst\u00e9mu bola preuk\u00e1zan\u00e1 aj v pr\u00edpade nemalobunkov\u00e9ho karcin\u00f3mu p\u013e\u00fac (NSCLC) (cut-off <\/em>hodnota 5 CTCs na 7,5 ml krvi)(24) a malobunkov\u00e9ho karcin\u00f3mu p\u013e\u00fac (SCLC), pri ktorom s\u00fa CTCs pr\u00edtomn\u00e9 vo vy\u0161\u0161om mno\u017estve ne\u017e v ostatn\u00fdch typoch n\u00e1dorov\u00fdch ochoren\u00ed (0 \u2013 44 896 buniek na 7,5 ml krvi s cut-off <\/em>hodnotou 50 CTCs na 7,5 ml krvi)(13).<\/p>\n

Okrem pou\u017eitia CTCs ako markerov progn\u00f3zy n\u00e1dorov\u00e9ho ochorenia m\u00f4\u017eu by\u0165 CTCs vyu\u017eit\u00e9 ako farmakodynamic<\/strong>k\u00e9 biomarkery<\/strong>. V reakcii na lie\u010dbu bol zaznamenan\u00fd pokles bu\u010f celkov\u00e9ho po\u010dtu CTCs(13,17,18,24) alebo ich subpopul\u00e1cie(25-27). V pr\u00edpade v\u00e4\u010d\u0161iny pacientov s SCLC, bolo podstatne zn\u00ed\u017een\u00e9 mno\u017estvo CTCs po jedinom cykle chemoterapie(13,28) v s\u00falade s vysokou mierou odpovede na lie\u010dbu na b\u00e1ze pla- tiny(29). Bolo preuk\u00e1zan\u00e9, \u017ee tieto zmeny s\u00fa nez\u00e1visle asociovan\u00e9 s dobrou progn\u00f3zou(13). Naopak, n\u00e1rast po\u010dtu CTCs po chemoterapii z priaznivej na nepriazniv\u00fa \u00farove\u0148 je spojen\u00fd s celkov\u00fdm krat\u0161\u00edm pre\u017e\u00edvan\u00edm pacientov s rakovinou prsn\u00edka, prostaty a CRC(17,18,30). \u010eal\u0161ie o\u010dak\u00e1van\u00e9 vyu\u017eitie CTC enumer\u00e1cie je v ranom \u0161t\u00e1diu vzniku rakoviny, ke\u010f pr\u00edtomnos\u0165 CTCs m\u00f4\u017ee pom\u00f4c\u0165 vybra\u0165 pacientov, u ktor\u00fdch je pravdepodobnos\u0165, \u017ee bud\u00fa ma\u0165 prospech z adjuvantnej lie\u010dby.<\/p>\n

CTCs ako predikt\u00edvny biomarker. <\/strong>V ide\u00e1lnom pr\u00edpade by CTCs bunky mohli by\u0165 vyu\u017eit\u00e9 ako tekut\u00e9 biopsie, ke\u010f by bol v\u00fdber \u0161pecifickej personalizovanej terapie uskuto\u010dnen\u00fd na z\u00e1klade molekul\u00e1rnych vlastnost\u00ed n\u00e1doru pacienta, zisten\u00fdch anal\u00fdzou CTCs. T\u00e1to sch\u00e9ma bola vysvetlen\u00e1 v \u0161t\u00fadii analyzuj\u00facich vzorky od pacientov s NSCLC pozit\u00edvnych na EGFR <\/em>mut\u00e1ciu, v ktorej bola u 19 z 20 pacientov identifikovan\u00e1 o\u010dak\u00e1van\u00e1 (na z\u00e1klade tkanivovej biopsie) aktivuj\u00faca EGFR <\/em>mut\u00e1cia(31). \u010eal\u0161\u00edm pr\u00edkladom je anal\u00fdza pre\u0161myku EML4-ALK, ktor\u00e1 sa vyskytuje u 3 \u2013 5 % pacientov s NSCLC. Pribli\u017ene 60 % pacientov m\u00e1 dobr\u00fa odpove\u010f na inhib\u00edciu ALK(32,33).\u00a0 V \u010fal\u0161\u00edch \u0161t\u00fadi\u00e1ch bola hodnoten\u00e1 expresia HER2 v CTCs v pr\u00edpade rakoviny prsn\u00edka(34) a expresia androg\u00e9nov\u00e9ho receptora v pr\u00edpade rakoviny prostaty(35,36), ktor\u00e9 by tie\u017e mohli by\u0165 potenci\u00e1lne vyu\u017eit\u00e9 ako predikt\u00edvne biomarkery. Niektor\u00e9 \u017eeny vykazovali rozdielny HER2 status v CTCs (pozit\u00edvny) a v prim\u00e1rnom n\u00e1dore (negat\u00edvny), pri\u010dom prosperovali z terapie trastuzumabom(37). Toto zistenie potvrdzuje n\u00e1zor, \u017ee vlastnosti CTC buniek s\u00fa relevantnej\u0161\u00edm terapeutick\u00fdm cie\u013eom, ke\u010f\u017ee tieto bunky s\u00fa zodpovedn\u00e9 za vznik metast\u00e1z.<\/p>\n

 <\/p>\n

Grantov\u00e1 podpora<\/h3>\n

T\u00e1to<\/em> publik\u00e1cia vznikla s podporou grantov Agent\u00fary na podporu v\u00fdskumu a v\u00fdvoja \u2013 APVV-14-0273 a APVV-14-0327.<\/em><\/p>\n

 <\/p>\n

 <\/p>\n

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

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    *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\u00face n\u00e1dorov\u00e9 bunky Cirkuluj\u00face n\u00e1dorov\u00e9 bunky (CTC) s\u00fa bunky uvo\u013e\u0148ovan\u00e9 do krvn\u00e9ho rie\u010diska z prim\u00e1rneho n\u00e1doru u\u017e v\u010dasne po\u010das jeho formovania a rastu a\/alebo jeho metast\u00e1zami. S\u00fa preuk\u00e1zate\u013ene asociovan\u00e9<\/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":[1037,1036,900],"class_list":["post-1588","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-cancer-metastasis","tag-circulating-tumour-cells","tag-liquid-biopsy","typ_clanku-review-article"],"acf":{"abstrakt":"

    Tumour cells have the ability of specific form of dedifferentiation, and thus they can acquire properties enabling their release from the primary tumour to circulation, where they can survive for a long time and become circulating tumour cells. These have the demonstrable potential to produce metastases, both in the form of single cells as well as their agglomerates. The possibility of their identification and quantification has, therefore, a significant prognostic value, which is based on already available and authority-approved methods also used in clinical routine, e. g. in patients with breast cancer. Moreover, there are ongoing several clinical trials that are likely to prove the predictive value of such analysis in patients with various oncological diseases and will contribute to the development of non-invasive tests working as part of the tests performed as so-called liquid biopsies.<\/p>\n

    Keywords:<\/strong> circulating tumour cells, cancer metastasis, 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":"

      \r\n \t
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    • Laboratory diagnostic possibilities for Clostridium difficile infections<\/li>\r\n \t
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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":"108","upload_clanok":{"ID":1586,"id":1586,"title":"Cirkuluj\u00face n\u00e1dorov\u00e9 bunky ako markery vyu\u017eite\u013en\u00e9 pri mana\u017emente","filename":"Cirkuluj\u00face-n\u00e1dorov\u00e9-bunky-ako-markery-vyu\u017eite\u013en\u00e9-pri-mana\u017emente.pdf","filesize":456785,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2018\/11\/Cirkuluj\u00face-n\u00e1dorov\u00e9-bunky-ako-markery-vyu\u017eite\u013en\u00e9-pri-mana\u017emente.pdf","link":"https:\/\/www.newslab.sk\/en\/cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-pacientov-s-nadorovymi-ochoreniami\/cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-2\/","alt":"","author":"7","description":"","caption":"","name":"cirkulujuce-nadorove-bunky-ako-markery-vyuzitelne-pri-manazmente-2","status":"inherit","uploaded_to":1588,"date":"2018-11-11 13:56:49","modified":"2018-11-11 13:56:49","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\/1588","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=1588"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1588\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1588"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1588"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1588"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}