{"id":1922,"date":"2020-05-05T15:34:06","date_gmt":"2020-05-05T13:34:06","guid":{"rendered":"https:\/\/www.newslab.sk\/molekulove-metody-detekcie-mutacii-v-kinazovej-domene-fuzneho-genu-bcr-abl1-u-pacientov-s-chronickou-myelocytovou-leukemiou\/"},"modified":"2020-05-05T15:39:26","modified_gmt":"2020-05-05T13:39:26","slug":"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","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/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\/","title":{"rendered":"Molecular detection methods of mutations in the kinase domain of fusion gene bcr-abl1 in patients with chronic myelocyte leukemia"},"content":{"rendered":"

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

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\u00davod<\/strong><\/h2>\n

Chronick\u00e1 myelocytov\u00e1 leuk\u00e9mia (CML) je klon\u00e1lne myeloproliferat\u00edvne ochorenie, ktor\u00e9 je v\u00fdsledkom neobmedzenej expanzie buniek v kostnej dreni. Diferenci\u00e1cia buniek sa zastavuje v \u0161t\u00e1diu blastov. Charakteristick\u00fdm znakom CML je pr\u00edtomnos\u0165 Philadelphia (Ph) chromoz\u00f3mu, ktor\u00fd vznik\u00e1 recipro\u010dnou translok\u00e1ciou medzi dlh\u00fdmi ramienkami chromoz\u00f3mov 9 a 22. Pri uvedenej translok\u00e1ci\u00ed t(9;22)(q34;q11) doch\u00e1dza k f\u00fazii g\u00e9nu BCR <\/em>na chromoz\u00f3me 22 s g\u00e9nom ABL1 <\/em>na chromoz\u00f3me 9. V\u00fdsledkom je vznik patologick\u00e9ho f\u00fazneho g\u00e9nu BCR-ABL1<\/em>(1). Nelie\u010den\u00e1 CML prebieha v troch f\u00e1zach, ktor\u00e9 ur\u010duj\u00fa progresiu ochorenia po\u010dtom trombocytov a blastov\u00fdch buniek v krvi a kostnej dreni. Prvotn\u00e9 \u0161t\u00e1dium CML predstavuje chronick\u00e1 f\u00e1za (angl. chronic phase, CP<\/em>), v ktorej je diagnostikovan\u00fdch a\u017e 90% pacientov. Po\u010das CP nezrel\u00e9 blastov\u00e9 bunky proliferuj\u00fa a diferencuj\u00fa pomaly, v d\u00f4sledku \u010doho m\u00f4\u017ee CP pretrv\u00e1va\u0165 mesiace a\u017e roky. Diagn\u00f3za CML sa stanov\u00ed hematologick\u00fdm a cytogenetick\u00fdm vy\u0161etren\u00edm a n\u00e1sledne sa za\u010dne lie\u010dba inhib\u00edtormi tyroz\u00ednkin\u00e1zy (TKI). V prvej l\u00ednii lie\u010dby v CP sa pacientovi v podmienkach SR m\u00f4\u017ee pod\u00e1va\u0165 bu\u010f imatinib, alebo nilotinib v z\u00e1vislosti od stratifik\u00e1cie pacienta pri stanoven\u00ed diagn\u00f3zy. Agres\u00edvnej\u0161ou formou CML je akcelerovan\u00e1 f\u00e1za (angl. accelerated phase, AP<\/em>), pri ktorej doch\u00e1dza k zv\u00fd\u0161enej prolifer\u00e1cii blastov\u00fdch buniek v perif\u00e9rnej krvi a kostnej dreni. V AP sa u pacientov m\u00f4\u017eu vyskytn\u00fa\u0165 pr\u00eddavn\u00e9 chromoz\u00f3mov\u00e9 aber\u00e1cie ako pr\u00edtomnos\u0165 extra chromoz\u00f3mu 8 alebo izochromoz\u00f3mu 17q (pozost\u00e1va z dvoch dlh\u00fdch ramienok 17. chromoz\u00f3mu), ktor\u00e9 s\u00fa pova\u017eovan\u00e9 za marker progresie ochorenia. Bez lie\u010dby progreduje AP do fat\u00e1lnej blastovej f\u00e1zy (angl. blastic phase, BP<\/em>). T\u00e1to f\u00e1za sa spr\u00e1va ako chemorezistentn\u00e1 ak\u00fatna leuk\u00e9mia. Pacientom v BP sa odpor\u00fa\u010da transplant\u00e1cia krvotvorn\u00fdch buniek, ktorej predch\u00e1dza kombinovan\u00e1 lie\u010dba intenz\u00edvnou chemoterapiou a TKI. Progn\u00f3za je v\u0161ak krajne nepriazniv\u00e1. Vzh\u013eadom na limitovan\u00fa \u00fa\u010dinnos\u0165 lie\u010dby pacientov v BP, je pr\u00ednosom v\u010dasn\u00fd z\u00e1chyt pr\u00eddavn\u00fdch zmien u rizikov\u00fdch pacientov(2).<\/p>\n

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\u00c9ra tyroz\u00ednkin\u00e1zov\u00fdch inhib\u00edtorov<\/h3>\n

Spo\u010diatku bola lie\u010dba CML zalo\u017een\u00e1 na pod\u00e1van\u00ed peror\u00e1lnej chemoterapie v podobe cytotoxick\u00fdch l\u00e1tok ako busulf\u00e1n alebo hydroxyurea. U v\u00e4\u010d\u0161iny lie\u010den\u00fdch pacientov v\u0161ak nedosiahla dostato\u010dn\u00fa lie\u010debn\u00fa odpove\u010f a takmer v\u0161etci pacienti progredovali do AP a BP. Od roku 1981 sa na lie\u010dbu CML za\u010dal pou\u017e\u00edva\u0165 interfer\u00f3n-alfa (IFN-\u03b1). Uk\u00e1zalo sa, \u017ee IFN-\u03b1 m\u00e1 priamy antiprolifera\u010dn\u00fd \u00fa\u010dinok a inhibuje patologick\u00fa sebaobnovu myeloidn\u00fdch progenitorov. \u0160tandardnou lie\u010deb- nou met\u00f3dou pre pacientov v CP a AP bola transplant\u00e1cia krvotvorn\u00fdch buniek, ktor\u00e1 v\u0161ak bola za\u0165a\u017een\u00e1 rizikom vysokej morbidity a mortality (napr. reakcia \u0161tepu proti hostite\u013eovi, GvHD). V\u00fdvoj malej molekuly imatinibmesyl\u00e1tu \u2013 tyroz\u00ednkin\u00e1zov\u00e9ho inhib\u00edtora sp\u00f4sobil v roku 2000 revol\u00faciu v lie\u010dbe CML. Bolo dok\u00e1zan\u00e9, \u017ee TKI silne interferuj\u00fa s interakciou medzi prote\u00ednom Bcr-Abl1 a adenoz\u00edntrifosf\u00e1tom (ATP). TKI sa via\u017ee v bl\u00edzkosti v\u00e4zobn\u00e9ho miesta ATP, a t\u00fdm sa stabilizuje tyroz\u00ednkin\u00e1za v inakt\u00edvnej konform\u00e1cii. Inhib\u00edciou tohto procesu sa zastav\u00ed aktiv\u00e1cia sign\u00e1lnych dr\u00e1h, ktor\u00e9 podporuj\u00fa leukemick\u00fd fenotyp bunky. Tyroz\u00ednkin\u00e1zy s\u00fa d\u00f4le\u017eit\u00fdmi medi\u00e1tormi sign\u00e1lnej kask\u00e1dy vo fyziologick\u00fdch bunk\u00e1ch. Ur\u010duj\u00fa k\u013e\u00fa\u010dov\u00e9 \u00falohy v r\u00f4znych biologick\u00fdch procesoch, ako je prolifer\u00e1cia, diferenci\u00e1cia, metabolizmus a apopt\u00f3za. Deregulovan\u00e1 aktivita tyroz\u00ednkin\u00e1zy je z\u00e1kladom pre patogen\u00e9zu \u013eudsk\u00fdch onkologick\u00fdch ochoren\u00ed.<\/p>\n

Imatinib predstavuje TKI prvej gener\u00e1cie. Medzi TKI 2. gener\u00e1cie patr\u00ed bosutinib, dasatinib a nilotinib. Ponatinib je TKI gener\u00e1cie. Cielen\u00e1 terapia vo forme selekt\u00edvnych TKI v\u00fdznamne zlep\u0161ila progn\u00f3zu, pred\u013a\u017eila celkov\u00e9 pre\u017e\u00edvanie aj kvalitu \u017eivota pacientov s CML(3).<\/p>\n

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Rezistencia proti TKI<\/h3>\n

Za chemorezistenciu n\u00e1dorov\u00fdch buniek je zodpovedn\u00fd cel\u00fd rad bunkov\u00fdch mechanizmov. Rozozn\u00e1vame dve kateg\u00f3rie rezistencie proti lie\u010dbe imatinibom, resp. TKI v\u0161eobecne \u2013 prim\u00e1rnu a sekund\u00e1rnu. Ak nie je zaznamenan\u00e1 odpove\u010f po inici\u00e1lnej lie\u010dbe, hovor\u00edme o prim\u00e1rnej rezistencii. V pr\u00edpade, ke\u010f momentu vzniku rezistencie predch\u00e1dza lie\u010debn\u00e1 odpove\u010f, ide o sekund\u00e1rnu rezistenciu. Prim\u00e1rna rezistencia (vn\u00fatorn\u00e1) sa vz\u0165ahuje na mechanizmy pr\u00edtomn\u00e9 v bunke sk\u00f4r, ako boli vystaven\u00e9 inhib\u00edtoru TK. Sekund\u00e1rna rezistencia (z\u00edskan\u00e1) je reprezentovan\u00e1 farmakologick\u00fdmi parametrami, ktor\u00e9 ovplyv\u0148uj\u00fa \u00fa\u010dinnos\u0165 lie\u010div molekul\u00e1rnymi zmenami. Pri sekund\u00e1rnej rezistencii doch\u00e1dza k strate lie\u010debnej odpovede. Bolo dok\u00e1zan\u00e9, \u017ee n\u00e1dorov\u00e9 mikroprostredie prispieva k vn\u00fatornej, ale aj z\u00edskanej rezistencii proti lie\u010div\u00e1m. Na mechanickej \u00farovni klasifikujeme rezistenciu proti TKI ako nez\u00e1visl\u00fa od Bcr-Abl1 alebo z\u00e1visl\u00fa od Bcr-Abl1. Z\u00e1visl\u00e1 rezistencia zah\u0155\u0148a mechanizmy, naj\u010dastej\u0161ie bodov\u00e9 mut\u00e1cie, ktor\u00e9 s\u00fa priamo z\u00e1visl\u00e9 od kin\u00e1zy Bcr-Abl1. Rezistencia nez\u00e1visl\u00e1 od Bcr-Abl1 je sprostredkovan\u00e1 r\u00f4znymi alternat\u00edvnymi sign\u00e1lnymi dr\u00e1hami v n\u00e1dorov\u00fdch bunk\u00e1ch. Klinick\u00e1 rezistencia je pozorovan\u00e1 prostredn\u00edctvom obidvoch mechanizmov, pri\u010dom z\u00edskan\u00e1 rezistencia je pravdepodobnej\u0161ie z\u00e1visl\u00e1 od Bcr-Abl1. Prim\u00e1rna rezistencia m\u00e1 tendenciu by\u0165 nez\u00e1visl\u00e1 od Bcr-Abl1. Rezistencia proti TKI je spojen\u00e1 s nepriazniv\u00fdmi klinick\u00fdmi v\u00fdsledkami (4).<\/p>\n

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Sekund\u00e1rna rezistencia<\/h3>\n

Mut\u00e1cie v kin\u00e1zovej dom\u00e9ne BCR-ABL1 <\/em>g\u00e9nu s\u00fa naj\u010dastej\u0161ou pr\u00ed\u010dinou sekund\u00e1rnej rezistencie proti lie\u010dbe TKI. Bolo op\u00edsan\u00fdch viac ako 100 mut\u00e1ci\u00ed v Abl1 tyroz\u00ednkin\u00e1zovej dom\u00e9ne. Najzn\u00e1mej\u0161ou mut\u00e1ciou u rezistentn\u00fdch pacientov je c.944C > T ved\u00faca k z\u00e1mene aminokysel\u00edn treon\u00ednu za izoleuc\u00edn v poz\u00edcii 315 prote\u00ednu Bcr-Abl1 (p. T315I), ktor\u00e1 sp\u00f4sobuje posun Abl1 kin\u00e1zy k akt\u00edvnej konform\u00e1cii. Naopak, mnoh\u00e9 bodov\u00e9 \u201emissense\u201c mut\u00e1cie Abl1 kin\u00e1zovej dom\u00e9ny pravdepodobne nemaj\u00fa pr\u00ed\u010dinn\u00fa s\u00favislos\u0165 so vznikom rezistencie, v niektor\u00fdch pr\u00edpadoch m\u00f4\u017ee by\u0165 tak\u00e1to mut\u00e1cia pr\u00edtomn\u00e1 aj pri dostato\u010dnej odpovedi na lie\u010dbu. Preto interpret\u00e1cia kauz\u00e1lnej s\u00favislosti rezistencie a mut\u00e1cie mus\u00ed vych\u00e1dza\u0165 z klinick\u00e9ho stavu a aktu\u00e1lnych laborat\u00f3rnych v\u00fdsledkov monitorovania.<\/p>\n

Bodov\u00e9 mut\u00e1cie v kin\u00e1zovej dom\u00e9ne BCR-ABL1 <\/em>g\u00e9nu mo\u017eno detegova\u0165 pomocou viacer\u00fdch met\u00f3d s r\u00f4znou \u00farov\u0148ou senzitivity aj \u010dasovej a manu\u00e1lnej n\u00e1ro\u010dnosti. V be\u017enej laborat\u00f3rnej praxi je najpou\u017e\u00edvanej\u0161ou met\u00f3dou priame sekvenovanie Abl1 kin\u00e1zovej dom\u00e9ny (senzitivita 10 \u2013 20%), alelovo \u0161pecifick\u00e1 PCR (ASO-PCR \u2013 allele specific oligonucleotide-polymerase chain reaction, senzitivita 0,01 \u2013 0,1%) a NGS sekvenovanie (sekvenovanie novej gener\u00e1cie, senzitivita 1%).<\/p>\n

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Mo\u017enosti detekcie pr\u00edtomnosti mut\u00e1ci\u00ed pomocou molekulov\u00fdch met\u00f3d<\/h3>\n

Sangerovo sekvenovanie<\/h4>\n

Sangerovo sekvenovanie je najroz\u0161\u00edrenej\u0161ia met\u00f3da pri rutinnom monitorovan\u00ed pacientov, ale m\u00e1 obmedzen\u00fa citlivos\u0165 detekcie 15 \u2013 20%. Pred samotn\u00fdm Sangerov\u00fdm sekvenovan\u00edm mus\u00ed prebehn\u00fa\u0165 amplifik\u00e1cia f\u00fazneho g\u00e9nu BCR<\/em>-ABL1 <\/em>pomocou nested polymer\u00e1zovej re\u0165azovej reakcie (PCR). V prvom kroku nested PCR sa amplifikuje cel\u00e1 kin\u00e1zov\u00e1 dom\u00e9na f\u00fazneho g\u00e9nu BCR-ABL1. <\/em>V druhom kroku nested PCR doch\u00e1dza k amplifik\u00e1cii fragmentu Abl1 kin\u00e1zy. Produkty PCR sa analyzuj\u00fa sekvenovan\u00edm. Princ\u00edpom Sangerovho sekvenovania je synt\u00e9za nov\u00e9ho re\u0165azca DNA pomocou DNA polymer\u00e1zy a inkorpor\u00e1cia termina\u010dn\u00e9ho dideoxynukleozidtrifosf\u00e1tu (ddNTP). Termin\u00e1ciou synt\u00e9zy vznikaj\u00fa r\u00f4zne dlh\u00e9 DNA fragmenty, ktor\u00e9 s\u00fa zakon\u010den\u00e9 konkr\u00e9tnym ddNTP. Po skon\u010den\u00ed sekvenovania prebehne elektroforetick\u00e1 separ\u00e1cia vzniknut\u00fdch DNA fragmentov na z\u00e1klade ich ve\u013ekosti (obr\u00e1zok 1)<\/em><\/strong>. V\u00fdsledn\u00e9 sekvencie sa porovnaj\u00fa s referen\u010dnou sekvenciou zdrav\u00e9ho jedinca. Reak\u010dn\u00e1 zmes Sangerovho sekvenovania mus\u00ed obsahova\u0165 templ\u00e1tov\u00fa (vy\u0161etrovan\u00fa) DNA, DNA polymer\u00e1zu, dNTP, ddNTP, \u0161pecifick\u00fd primer a reak\u010dn\u00fd pufor.<\/p>\n

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Alelovo\u0161pecifick\u00e1 PCR<\/h4>\n

Alelovo\u0161pecifick\u00e1 PCR\u00a0 (AS-PCR)\u00a0 poskytuje\u00a0 inform\u00e1ciu o pr\u00edtomnosti alebo nepr\u00edtomnosti mut\u00e1cie na jednej alele g\u00e9nu. Na stanovenie genotypu je nutn\u00e9, aby prebehli dve PCR reakcie, jedna sl\u00fa\u017ei len na amplifik\u00e1ciu alely bez mut\u00e1cie, druh\u00e1 reakcia je \u0161pecifick\u00e1 pre mutantn\u00fa alelu (obr\u00e1zok 2)<\/em><\/strong>. Pr\u00edslu\u0161n\u00e9 PCR primery s\u00fa sekven\u010dne identick\u00e9, odli\u0161uj\u00fa sa len v nukleotide na 3\u00b4 konci, \u010d\u00edm sa zabezpe\u010d\u00ed preferen\u010dn\u00e1 amplifik\u00e1cia jednej alely pred druhou. Na vyl\u00fa\u010denie nespr\u00e1vnej genotypiz\u00e1cie v d\u00f4sledku zlyhania PCR sa do ka\u017edej z reakci\u00ed prid\u00e1va \u010fal\u0161\u00ed p\u00e1r alelovone\u0161pecifick\u00fdch primerov pre amplifik\u00e1ciu kontroln\u00e9ho fragmentu. AS-PCR pon\u00faka v\u00fdhody v podobe r\u00fdchlej\u0161\u00edch v\u00fdsledkov anal\u00fdzy, ni\u017e\u0161\u00edch n\u00e1kladov a \u013eah\u0161ieho pracovn\u00e9ho protokolu. Jej detek\u010dn\u00fd limit je 0,5 \u2013 1% mutovanej alely(5). Nev\u00fdhodou met\u00f3dy je, \u017ee je schopn\u00e1 detegova\u0165 len konkr\u00e9tnu mut\u00e1ciu.<\/p>\n

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NGS \u2013 sekvenovanie novej gener\u00e1cie<\/h4>\n

NGS (sekvenovanie novej gener\u00e1cie) umo\u017e\u0148uje detegova\u0165 a kvantifikova\u0165 variant v Bcr-Abl1 transkriptoch a\u017e na \u00farovni 1%. NGS pozost\u00e1va z troch krokov. V prvom kroku doch\u00e1dza k vytvoreniu sekvena\u010dnej kni\u017enice fragment\u00e1ciou genomickej DNA. V druhom kroku sa sekvena\u010dn\u00e1 kni\u017enica amplifikuje. V tre\u0165om kroku doch\u00e1dza k mas\u00edvnemu paraleln\u00e9mu sekvenovaniu ka\u017edej jednej molekuly DNA v \u0161peci\u00e1lnom sekven\u00e1tore (obr\u00e1zok 3). <\/em><\/strong>Presnos\u0165 NGS met\u00f3dy z\u00e1vis\u00ed od po\u010dtu \u010d\u00edtan\u00ed ka\u017ed\u00e9ho fragmentu DNA. U pacientov so zlyhan\u00edm na lie\u010dbe TKI sa m\u00f4\u017eu vyskytn\u00fa\u0165 mut\u00e1cie pod detek\u010dn\u00fdm limitom Sangerovho sekvenovania, ktor\u00e9 sa rutinne vyu\u017e\u00edva vo v\u00e4\u010d\u0161ine laborat\u00f3ri\u00ed. NGS met\u00f3da m\u00f4\u017ee by\u0165 u\u017eito\u010dn\u00e1 u pacientov, ktor\u00ed nezlep\u0161uj\u00fa svoje odpovede na terapiu druhou, resp. tre\u0165ou l\u00edniou TKI. U pacientov v AP a BC je \u010dasto detegovan\u00fdch viac mut\u00e1ci\u00ed, ke\u010f met\u00f3da NGS m\u00f4\u017ee poskytn\u00fa\u0165 jednoduch\u00fd sp\u00f4sob identifik\u00e1cie viacer\u00fdch mut\u00e1ci\u00ed u jedn\u00e9ho pacienta(6).<\/p>\n

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Z\u00e1ver<\/h3>\n

Cielen\u00e1 lie\u010dba pomocou TKI v\u00fdrazne zlep\u0161ila progn\u00f3zu aj kvalitu \u017eivota pacientov s CML. Probl\u00e9mom terapie v\u0161ak zost\u00e1va rezistencia, preto je potrebn\u00e9 pozna\u0165 biologick\u00fa podstatu rezistencie a prisp\u00f4sobi\u0165 jej lie\u010debn\u00fa strat\u00e9giu. Potvrdenie mut\u00e1cie v kin\u00e1zovej dom\u00e9ne ako pr\u00ed\u010diny rezistencie m\u00e1 ve\u013ek\u00fd klinick\u00fd v\u00fdznam, preto\u017ee umo\u017e\u0148uje v\u010das ovplyvni\u0165 neefekt\u00edvnu terapiu. Pod\u013ea typu zistenej mut\u00e1cie doch\u00e1dza k zv\u00fd\u0161eniu d\u00e1vky pod\u00e1van\u00e9ho lie\u010diva alebo k zmene lie\u010dby. V pr\u00edpade rezistencie proti imatinibu mo\u017enos\u0165 \u00faspe\u0161nej terapie prin\u00e1\u0161aj\u00fa TKI 2. gener\u00e1cie, resp. 3. gener\u00e1cie, niektor\u00e9 z nich u\u017e v s\u00fa\u010dasnosti zast\u00e1vaj\u00fa miesto aj v prvol\u00edniovej lie\u010dbe CML. V ojedinel\u00fdch pr\u00edpadoch do \u00favahy prich\u00e1dza aj alog\u00e9nna transplant\u00e1cia krvotvorn\u00fdch buniek. \u00dalohou molekulovej genetiky je presn\u00e1 a r\u00fdchla detekcia mut\u00e1ci\u00ed aj \u010fal\u0161ie monitorovanie v\u00fdvoja mutovan\u00e9ho leukemick\u00e9ho klonu.<\/p>\n

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

    \n
  1. Thompson PA, Kantarjian HM, Cortes J Diagnosis and Treatment of Chronic Myeloid Leukemia in 2015. Mayo Clinic Procceedings 2015; 90(10): 1440-1454.<\/li>\n
  2. Gong Z, Medeiros , Cortes, JE, et al. Cytogenetics-based risk prediction of blastic transformation of chronic myeloid leukemia in the era of TKI therapy. Blood Advances 2017; 1(26): 2541-2552.<\/li>\n
  3. Jabbour Chronic myeloid leukemia: First-line drug of choice. American Journal of Hematology 2015; 91(1): 59-66.<\/li>\n
  4. Jones D, Kamel-Reid S, Bahler D, et Laboratory Practice Guidelines for Detecting and Reporting BCR-ABL Drug Resistance Mutations in Chronic Myelogenous Leukemia and Acute Lymphoblastic Leukemia. Journal of Molecular Diagnostics 2009; 11: 4-11.<\/li>\n
  5. Kockan B, Toptas T, Atik\u00fcnd\u00fcz Tuglular AT, et Molecular screening and the clinical impacts of BCR-ABL KD mutations in patients with imatinib-resistant chroncic myeloid leukemia. Oncology Letters 2018; 15(2): 2419-2424.<\/li>\n
  6. Soverini S, Abruzzese E, Bocchia M, et Next-generation sequencing for BCR-ABL1 kinase doamin mutation testing in patients with chronic myeloid leukemia: a position paper. Jounal of Hematology & Oncology 2019; 12: 131.<\/li>\n
  7. Arora Next-Generation Sequencing and Its Application: Empowering in Public Health Beyond Reality. Microbial Technology for the Welfare of Society 2019; (313-341), ISBN 978-981-13-8843-9.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    *All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper.   \u00davod Chronick\u00e1 myelocytov\u00e1 leuk\u00e9mia (CML) je klon\u00e1lne myeloproliferat\u00edvne ochorenie, ktor\u00e9 je v\u00fdsledkom neobmedzenej expanzie buniek v kostnej dreni. Diferenci\u00e1cia buniek sa zastavuje v \u0161t\u00e1diu blastov. Charakteristick\u00fdm znakom CML je<\/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":[1395,720,1396],"class_list":["post-1922","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-mutations-in-the-abl1-kinase-domain","tag-next-generation-sequencing-ngs","tag-sanger-sequencing","typ_clanku-review-article"],"acf":{"abstrakt":"

    The implementation of the treatment with tyrosine kinase inhibitors (TKIs) in patients with chronic myelocyte leukemia has significantly influenced the prognosis of the disease. TKI treatment is a significant benefit, but 25- 35% of patients need to change treatment within 5 years due to resistance or treatment failure. The most common mechanism responsible for resistance is the development of mutations in the kinase domain. Sanger sequencing is considered the standard method for screening mutations in the BCR-ABL1 <\/em>fusion gene. Currently, most work is being done to implement the next-generation sequencing technology as a standard method.<\/p>\n

    Keywords: <\/strong>mutations in the Abl1 kinase domain, Sanger sequencing, next-generation sequencing (NGS)<\/strong><\/p>\n","casopis":[{"ID":1893,"post_author":"7","post_date":"2020-05-05 11:32:54","post_date_gmt":"2020-05-05 09:32:54","post_content":"

      \r\n \t
    • Identification of metabolic pathways in pathogenesis of diabetic retinopathy using exome sequencing \u2013 a pilot study<\/li>\r\n \t
    • Anti-tumour effects of vitamin D<\/li>\r\n \t
    • Molecular detection methods of mutations in the kinase domain of fusion gene bcr-abl1 in patients with chronic myelocyte leukemia<\/li>\r\n \t
    • The case report of toxoplasmic meningoencephalitis with fatal outcome in HIV patient<\/li>\r\n \t
    • Carcinosarcoma-like endometrioid carcinoma of the uterus: case report of rare non-high-grade tumor<\/li>\r\n<\/ul>","post_title":"newsLab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-4","to_ping":"","pinged":"","post_modified":"2020-05-05 15:13:41","post_modified_gmt":"2020-05-05 13:13:41","post_content_filtered":"","post_parent":0,"guid":"https:\/\/www.newslab.sk\/?post_type=casopis&p=1893","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"37-40","upload_clanok":{"ID":1920,"id":1920,"title":"NEWSLAB_1-2020_Bezecn\u00e1","filename":"NEWSLAB_1-2020_Bezecn\u00e1.pdf","filesize":228161,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2020\/05\/NEWSLAB_1-2020_Bezecn\u00e1.pdf","link":"https:\/\/www.newslab.sk\/en\/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\/newslab_1-2020_bezecna-2\/","alt":"","author":"7","description":"","caption":"","name":"newslab_1-2020_bezecna-2","status":"inherit","uploaded_to":1922,"date":"2020-05-05 13:21:30","modified":"2020-05-05 13:21:30","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\/1922","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=1922"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1922\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1922"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1922"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1922"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}