{"id":1533,"date":"2018-11-09T10:40:07","date_gmt":"2018-11-09T09:40:07","guid":{"rendered":"http:\/\/www.newslab.sk\/2018\/11\/09\/geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-syndromom-a-sposoby-ich-detekcie\/"},"modified":"2018-11-12T08:24:02","modified_gmt":"2018-11-12T07:24:02","slug":"geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-syndromom-a-sposoby-ich-detekcie","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-syndromom-a-sposoby-ich-detekcie\/","title":{"rendered":"Genetically important aberrations in patients with myelodysplastic syndrome and laboratory methods of their detection"},"content":{"rendered":"

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

 <\/p>\n

\u00davod<\/strong>
\nMyelodysplastick\u00fd syndr\u00f3m (MDS) predstavuje heterog\u00e9nnu skupinu chronick\u00fdch myeloidn\u00fdch ochoren\u00ed s p\u00f4vodom v kostnej dreni. Ide o klon\u00e1lnu poruchu krvotvorby ako d\u00f4sledok mut\u00e1ci\u00ed vznikaj\u00facich v pluripotentn\u00fdch kme\u0148ov\u00fdch bunk\u00e1ch. Vplyvom mut\u00e1ci\u00ed doch\u00e1dza k nedostato\u010dnej matur\u00e1cii, diferenci\u00e1cii, resp. poru\u0161enej prolifer\u00e1cii, a k morfologickej dyspl\u00e1zii krvn\u00fdch elementov. Tieto zmeny ved\u00fa k vzniku inefekt\u00edvnej dysplastickej hematopo\u00e9zy s potenci\u00e1lnym rizikom transform\u00e1cie MDS do ak\u00fatnej myeloblastovej leuk\u00e9mie (AML) a\u017e v 30 % v\u0161etk\u00fdch pr\u00edpadov. Spolo\u010dn\u00fdm znakom ochorenia je hypercelul\u00e1rna kostn\u00e1 dre\u0148 a pr\u00edtomnos\u0165 r\u00f4zneho stup\u0148a perif\u00e9rnej cytop\u00e9nie. Typick\u00e1 je pr\u00edtomnos\u0165 an\u00e9mie, ale aj leukop\u00e9nie a trombocytop\u00e9nie a ich vz\u00e1jomn\u00e1 kombin\u00e1cia(1). MDS patr\u00ed medzi \u010dast\u00e9 hematologick\u00e9 malignity v Eur\u00f3pe s celkov\u00fdm v\u00fdskytom pribli\u017ene 4 pr\u00edpady na 100 000 obyvate\u013eov za rok. V\u00fdskyt prudko st\u00fapa so zvy\u0161uj\u00facim sa vekom najm\u00e4 vo vekovej skupine nad 60 rokov a vo veku \u2265 80 rokov je incidencia ochorenia a\u017e 50 pr\u00edpadov\/100 000 obyvate\u013eov ro\u010dne. Medi\u00e1n veku pri n\u00e1stupe choroby je okolo 70 rokov, pri\u010dom z celkov\u00e9ho po\u010dtu pacientov len 10 % tvoria pacienti s vekom pod 50 rokov(2). Deti s\u00fa postihnut\u00e9 zriedkavo, priebeh ochorenia a cytogenetick\u00e9 n\u00e1lezy s\u00fa odli\u0161n\u00e9 od dospel\u00fdch pacientov(3).<\/p>\n

Pr\u00ed\u010diny vzniku MDS<\/h3>\n

V\u00fdvoj MDS je viacstup\u0148ov\u00fd proces, pri ktorom doch\u00e1dza k zmen\u00e1m v genetickom materi\u00e1li pluripotentnej hematopo- etickej kme\u0148ovej bunky. T\u00e1 je morfologicky a funk\u010dne odli\u0161n\u00e1 a z\u00edskava rastov\u00fa v\u00fdhodu. Mechanizmus patogen\u00e9zy MDS nie je v s\u00fa\u010dasnosti \u00faplne objasnen\u00fd, predpoklad\u00e1 sa postihnutie g\u00e9nov reguluj\u00facich rast, diferenci\u00e1ciu a z\u00e1nik bunky. Takto pozmenen\u00e1 bunka proliferuje a postupne jej klony nahr\u00e1dzaj\u00fa zdrav\u00e9 kme\u0148ov\u00e9 bunky, \u010d\u00edm nast\u00e1va \u00fatlm alebo zastavenie norm\u00e1lnej krvotvorby(4). Rozozn\u00e1vame dva typy MDS \u2013 prim\u00e1rny a sekund\u00e1rny. O prim\u00e1rnom hovor\u00edme, ke\u010f nie je zn\u00e1ma pr\u00ed\u010dina ochorenia. Predstavuje 85 % zo v\u0161etk\u00fdch MDS a postihuje hlavne star\u0161iu popul\u00e1ciu s priemern\u00fdm vekom 60-70 rokov. V posledn\u00fdch rokoch st\u00fapa incidencia prim\u00e1rneho MDS, a to vzh\u013eadom na lep\u0161ie diagnostick\u00e9 met\u00f3dy, starnutie popul\u00e1cie a p\u00f4sobenie faktorov prostredia(5). Pri sekund\u00e1rnom MDS (~ 15 %) vieme objasni\u0165 pr\u00ed\u010diny vzniku ochorenia. M\u00f4\u017ee ich by\u0165 viacero, napr\u00edklad chemoterapia, r\u00e1dioterapia alebo vplyv environment\u00e1lnych tox\u00ednov(6).<\/p>\n

 <\/p>\n

Progn\u00f3za ochorenia<\/h3>\n

Priebeh ochorenia je ve\u013emi variabiln\u00fd, od naj\u013eah\u0161\u00edch foriem s priemern\u00fdm pre\u017e\u00edvan\u00edm nieko\u013eko rokov a\u017e po naj\u0165a\u017e\u0161ie formy s pre\u017e\u00edvan\u00edm ni\u017e\u0161\u00edm ako 5 mesiacov(7). Z h\u013eadiska progn\u00f3zy existuje nieko\u013eko prognostick\u00fdch syst\u00e9mov, ktor\u00e9 na z\u00e1klade s\u00favisiacich znakov triedia pacientov do rizikov\u00fdch kateg\u00f3ri\u00ed(8). V s\u00fa\u010dasnosti sa pou\u017e\u00edva Revidovan\u00fd <\/strong>medzin\u00e1rodn\u00fd prognostick\u00fd sk\u00f3rovac\u00ed syst\u00e9m \u2013 IPSS-R<\/strong>, ktor\u00fd del\u00ed pacientov do 5 rizikov\u00fdch kateg\u00f3ri\u00ed v s\u00falade s konkr\u00e9tnymi parametrami\u00a0 ud\u00e1vaj\u00facimi\u00a0 prognostick\u00fd\u00a0 v\u00fdznam a pod\u013ea ktor\u00fdch prebieha lie\u010dba. IPSS-R zah\u0155\u0148a aj MDS Cy<\/strong>togenetick\u00fd sk\u00f3rovac\u00ed syst\u00e9m (tabu\u013eka 1)<\/em><\/strong>, ktor\u00fd ur\u010duje prognostick\u00fa skupinu pacienta na z\u00e1klade cytogenetick\u00fdch n\u00e1lezov v karyotype(9).<\/p>\n

 <\/p>\n

Chromoz\u00f3mov\u00e9 aber\u00e1cie<\/h3>\n

Chromoz\u00f3mov\u00e9 aber\u00e1cie zohr\u00e1vaj\u00fa d\u00f4le\u017eit\u00fa \u00falohu v patogen\u00e9ze, progn\u00f3ze a diagnostike ochorenia. Pri prim\u00e1rnom MDS sa zaznamenali pri 40 \u2013 60 % pacientoch. Pacienti so sekund\u00e1rnym MDS maj\u00fa spravidla viac zmien v genotype, a\u017e v 80 \u2013 90 % s\u00fa pozorovan\u00e9 chromoz\u00f3mov\u00e9 prestavby (6). Komplexn\u00fd karyotyp s viac ako tromi aber\u00e1ciami naraz je pozorovan\u00fd pribli\u017ene u 15 % de novo <\/em>MDS oproti 50 % pri sekund\u00e1rnom MDS(10).<\/p>\n

Medzi naj\u010dastej\u0161ie zmeny patria del(5q)\/\u20135, 01507\/ del(7q), +8, del(20q) <\/strong>a \u2013Y<\/strong>. Menej \u010dasto sa vyskytuje \u201317\/ del(17p)\/i(17q), \u201318\/del(18q), +21, +19, inv\/t\/del(3q), \u201313\/ del(13q), \u201321, t(5q), +11, +1\/+1q, del(12p), del(11q), t(7q), +mar a in\u00e9(7,11).<\/p>\n

Pod\u013ea pr\u00edtomn\u00fdch zmien v bunk\u00e1ch m\u00f4\u017eeme predpoklada\u0165 progn\u00f3zu ochorenia. U pacientov s dobrou progn\u00f3zou nach\u00e1dzame norm\u00e1lny karyotyp, pr\u00edpadne len jednoduch\u00e9 zmeny. Priebeh ochorenia je mierny, dlhotrvaj\u00faci s pre\u017e\u00edvan\u00edm nieko\u013eko rokov a lie\u010dba je v\u00e4\u010d\u0161inou symptomatick\u00e1. \u010c\u00edm je progn\u00f3za hor\u0161ia, t\u00fdm je krat\u0161\u00ed \u010das pre\u017e\u00edvania pacientov, s\u00fa pr\u00edtomn\u00e9 r\u00f4zne genetick\u00e9 zmeny a je v\u00e4\u010d\u0161ia aj pravdepodobnos\u0165 leukemickej transform\u00e1cie(12).<\/p>\n

 <\/p>\n

Sp\u00f4soby detekcie genetick\u00fdch zmien<\/h3>\n

1.\u00a0 Cytogenetick\u00e1 anal\u00fdza<\/h4>\n

K z\u00e1kladn\u00fdm met\u00f3dam na detekciu aber\u00e1ci\u00ed patr\u00ed cytogenetick\u00e1 anal\u00fdza chromoz\u00f3mov. Umo\u017e\u0148uje sledova\u0165 numerick\u00e9 a \u0161trukt\u00farovan\u00e9 prestavby karyotypu. Ve\u013ekou v\u00fdhodou met\u00f3dy je mo\u017enos\u0165 vy\u0161etri\u0165 v\u0161etky chromoz\u00f3my v jednom poh\u013eade. Nev\u00fdhodou je neschopnos\u0165 identifikova\u0165 submikroskopick\u00e9 zmeny pod detek\u010dn\u00fdm limitom met\u00f3dy a z\u00e1rove\u0148 potreba metaf\u00e1z.<\/p>\n

 <\/p>\n

2.\u00a0 FISH met\u00f3da<\/h4>\n

Fluorescen\u010dn\u00e1 in situ hybridiz\u00e1cia (FISH) je be\u017enou met\u00f3dou v cytogenetick\u00fdch laborat\u00f3ri\u00e1ch a k jej v\u00fdhod\u00e1m patr\u00ed mo\u017enos\u0165 anal\u00fdzy buniek nielen v metaf\u00e1ze, ale aj v interf\u00e1ze, ke\u010f nie je nutn\u00e1 kultiv\u00e1cia buniek. \u010eal\u0161ou v\u00fdhodou je r\u00fdchle analyzovanie ve\u013ek\u00e9ho mno\u017estva buniek. Z\u00e1rove\u0148 umo\u017e\u0148uje odhali\u0165 aber\u00e1cie aj asi v 15 % cytogeneticky negat\u00edvnych pr\u00edpadov a pom\u00e1ha objas\u0148ova\u0165 komplexn\u00e9 prestavby. Limituj\u00facim faktorom met\u00f3dy je mo\u017enos\u0165 identifik\u00e1cie len t\u00fdch \u00fasekov DNA, pri ktor\u00fdch s\u00fa pou\u017eit\u00e9 sondy. Na relevantn\u00fa detekciu treba ma\u0165 vo vzorke aspo\u0148 1 \u2013 5 % aberantn\u00fdch interf\u00e1zov\u00fdch jadier.<\/p>\n

 <\/p>\n

Tabu\u013eka 1. <\/em><\/strong>MDS Cytogenetick\u00fd sk\u00f3rovac\u00ed syst\u00e9m<\/em>(9, upraven\u00e9)<\/em><\/p>\n\n\n\n\n\n\n\n\n
Prognostick\u00e1 skupina<\/strong><\/td>\nCytogenetick\u00e9 abnormality<\/strong><\/td>\nMedi\u00e1n pre\u017e\u00edvania<\/strong><\/p>\n

(roky)<\/strong><\/td>\n

Medi\u00e1n progresie<\/strong><\/p>\n

do AML (roky)<\/strong><\/td>\n<\/tr>\n

Ve\u013emi dobr\u00e1<\/strong><\/td>\n\u2013Y, del(11q)<\/td>\n5,4<\/td>\nnedosiahnut\u00fd<\/td>\n<\/tr>\n
Dobr\u00e1<\/strong><\/td>\nnorm\u00e1lny karyotyp, del(5q), del(12p), del(20q), dvojit\u00e9 zah\u0155\u0148aj\u00face del(5q)<\/td>\n4,8<\/td>\n9,4<\/td>\n<\/tr>\n
Stredn\u00e1<\/strong><\/td>\ndel(7q), +8, +19, i(17q), ak\u00e9ko\u013evek in\u00e9 samostatn\u00e9 alebo dvojit\u00e9 nez\u00e1visl\u00e9 klony<\/td>\n2,7<\/td>\n2,5<\/td>\n<\/tr>\n
Zl\u00e1<\/strong><\/td>\n\u20137, inv(3)\/t(3q)\/del(3q), dvojit\u00e1 zah\u0155naj\u00faca \u20137\/del(7q), komplex 3 abnormal\u00edt<\/td>\n1,5<\/td>\n1,7<\/td>\n<\/tr>\n
Ve\u013emi zl\u00e1<\/strong><\/td>\nkomplex > 3 abnormal\u00edt<\/td>\n0,7<\/td>\n0,7<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

3.\u00a0 MLPA anal\u00fdza<\/h4>\n

Podobne ako FISH aj molekul\u00e1rno-cytogenetick\u00e1 met\u00f3da MLPA (Multiplex ligation-dependent probe amplification) vyu\u017e\u00edva naviazanie oligonukleotidov\u00fdch sond na cie\u013eov\u00fa sekvenciu DNA na z\u00e1klade ich komplementarity. Oproti FISH je t\u00e1to met\u00f3da schopn\u00e1 identifikova\u0165 naraz viac ako 40 r\u00f4znych sekvenci\u00ed DNA v jednej reakcii(13). Sondy vyu\u017e\u00edvan\u00e9 pri MLPA met\u00f3de maj\u00fa krat\u0161iu d\u013a\u017eku (50 \u2013 70 bp) a umo\u017e\u0148uj\u00fa citliv\u00fa detekciu aj in\u00fdch klinicky relevantn\u00fdch MDS abnormal\u00edt s ni\u017e\u0161ou frekvenciou. V\u00fdhodou MLPA je odl\u00ed\u0161enie sekvenci\u00ed, ktor\u00e9 sa navz\u00e1jom l\u00ed\u0161ia rozdielom jedn\u00e9ho nukleotidu. Nev\u00fdhodou techniky je potreba ma\u0165 vo vzorke aspo\u0148 30 % mutovan\u00fdch buniek na spo\u013eahliv\u00fa detekciu abnormal\u00edt(14).<\/p>\n

 <\/p>\n

Cie\u013eom na\u0161ej pr\u00e1ce bolo:<\/h3>\n
    \n
  1. Vytvori\u0165 a analyzova\u0165 s\u00fabor pacientov s diagn\u00f3zou MDS vy\u0161etren\u00fdch na na\u0161om oddelen\u00ed genetiky v \u010dase od febru\u00e1ra 2016 do apr\u00edla<\/li>\n
  2. Zisti\u0165 pr\u00edtomnos\u0165 chromoz\u00f3mov\u00fdch aber\u00e1ci\u00ed, charakterizova\u0165 ich, ur\u010di\u0165 ich incidenciu v s\u00fabore a rozdeli\u0165 pacientov pod\u013ea pr\u00edtomn\u00fdch aber\u00e1ci\u00ed.<\/li>\n
  3. Zhodnoti\u0165 vz\u0165ah medzi v\u00fdsledkami vy\u0161etren\u00ed a progn\u00f3zou.<\/li>\n
  4. Stanovi\u0165 pr\u00ednos jednotliv\u00fdch met\u00f3d pri hodnoten\u00ed chromoz\u00f3mov\u00fdch aber\u00e1ci\u00ed a porovnanie \u00faspe\u0161nosti pou\u017eit\u00fdch met\u00f3d \u2013 klasick\u00e1 cytogenetika, met\u00f3da FISH a met\u00f3da<\/li>\n<\/ol>\n

     <\/p>\n

    S\u00fabor pacientov a met\u00f3dy<\/h3>\n

    Do pr\u00e1ce sme zaradili pacientov s diagn\u00f3zou MDS, ktor\u00ed boli vy\u0161etrovan\u00ed v na\u0161om laborat\u00f3riu genetiky od febru\u00e1ra 2016 do apr\u00edla 2018. Krit\u00e9riom na zaradenie pacienta bola diagn\u00f3za stanoven\u00e1 lek\u00e1rom na \u017eiadanke. Celkovo sme analyzovali 620 vzoriek pacientov, z toho 383 vzoriek kostnej drene (KD) a 237 vzoriek perif\u00e9rnej krvi (PK). V pr\u00e1ci sme pou\u017eili nasledovn\u00e9 met\u00f3dy: cytogenetika, FISH a MLPA.<\/p>\n

    Prepar\u00e1ty na vy\u0161etrenie karyotypu boli pripraven\u00e9 24-hodinovou kultiv\u00e1ciou buniek KD v kompletnom m\u00e9diu a spracovan\u00e9 pod\u013ea \u0161tandardn\u00fdch postupov. U 360 pacientov bolo analyzovan\u00fdch, ak to bolo mo\u017en\u00e9, 20 metaf\u00e1z a bol stanoven\u00fd cytogenetick\u00fd z\u00e1pis pod\u013ea ISCN (International System of Chromosome Nomenclature). Pri met\u00f3de FISH sa ako vstupn\u00fd materi\u00e1l pou\u017eila kultivovan\u00e1 KD a\/alebo PK pacienta pripraven\u00e1 priamym spracovan\u00edm. N\u00e1sledne prebehla hybridiz\u00e1cia s pr\u00edslu\u0161nou sondou a hodnotenie prepar\u00e1tov pod fluorescen\u010dn\u00fdm mikroskopom. Spolu bolo zhodnoten\u00fdch 521 vzoriek, 380 KD a 141 PK. Vstupn\u00fdm materi\u00e1lom na anal\u00fdzu MLPA bola izolovan\u00e1 DNA zo vzorky KD a\/alebo PK dodan\u00e1 v EDTA. Na anal\u00fdzy sa pou\u017eil kit SALSA MLPA P414-B1 MDS probemix (MRC \u2013 Holland) na detekciu 46 \u0161pecifick\u00fdch chromoz\u00f3mov\u00fdch oblast\u00ed s amplifika\u010dn\u00fdmi produktmi od 122 do 469 nukleotidov. Sondy s\u00fa navrhnut\u00e9 na naj- roz\u0161\u00edrenej\u0161ie a na prognosticky relevantn\u00e9 aber\u00e1cie pri MDS pod\u013ea IPSS-R. Toto met\u00f3dou sa vy\u0161etrilo 398 pacientov (214 KD a 184 PK).<\/p>\n

     <\/p>\n

    V\u00fdsledky<\/h3>\n

    1.\u00a0 Charakteristika a anal\u00fdza s\u00faboru pacientov<\/h4>\n

    S\u00fabor zah\u0155\u0148a vzorky kostnej drene alebo perif\u00e9rnej krvi od 620 pacientov a pozost\u00e1va z 289 (46,6 %) pacientov mu\u017esk\u00e9ho a 331 (53,4 %) \u017eensk\u00e9ho pohlavia. Vekov\u00fd medi\u00e1n pacientov v \u010dase ur\u010denia diagn\u00f3zy bol 69 rokov s vekov\u00fdm rozp\u00e4t\u00edm 13 a\u017e 93 rokov. Pacienti vy\u0161etren\u00ed zo vzorky kostnej drene mali patol\u00f3gie v 36 % pr\u00edpadoch, pacienti vy\u0161etren\u00ed zo vzorky perif\u00e9rnej krvi len v 14 % pr\u00edpadoch.<\/p>\n

     <\/p>\n

    1.\u00a0 Incidencia chromoz\u00f3mov\u00fdch abnormal\u00edt<\/h4>\n

    Z v\u00fdsledkov vzoriek kostnej drene 383 pacientov, ktor\u00ed boli vy\u0161etren\u00ed cytogeneticky, met\u00f3dou FISH a\/alebo MLPA, sme charakterizovali a n\u00e1sledne stanovili v\u00fdskyt naj\u010dastej\u0161\u00edch abnormal\u00edt. Detegovali sme spolu 287 chromoz\u00f3mov\u00fdch aber\u00e1ci\u00ed, pri\u010dom u 99 % pacientov boli pou\u017eit\u00e9 aspo\u0148 dve met\u00f3dy na vy\u0161etrenie. Na grafe\u00a0<\/em><\/strong>je zn\u00e1zornen\u00e9 jednotliv\u00e9 zast\u00fapenie aber\u00e1ci\u00ed ako izolovan\u00e9 abnormality v gen\u00f3me a ako abnormality, ktor\u00e9 sa vyskytli v kombin\u00e1cii s in\u00fdmi patol\u00f3giami.<\/p>\n

     <\/p>\n

    2.\u00a0 Ur\u010denie progn\u00f3zy pacientov<\/h4>\n

    Pomocou v\u00fdsledkov vy\u0161etren\u00ed z anal\u00fdz vzoriek kostnej drene z\u00edskan\u00fdch met\u00f3dami cytogenetiky, FISH a\/alebo MLPA bola stanoven\u00e1 progn\u00f3za a medi\u00e1n pre\u017e\u00edvania 383 pacientom. Hodnoten\u00e9 bolo zast\u00fapenie jednotliv\u00fdch pacientov v prognostick\u00fdch skupin\u00e1ch a ich celkov\u00e9 pre\u017e\u00edvanie.<\/p>\n

    Medi\u00e1n pre\u017e\u00edvania sme vypo\u010d\u00edtali \u0161tatistickou Kaplanovou-Meierovou met\u00f3dou spolo\u010dne pre 351 pacientov, od ktor\u00fdch sme mali dostupn\u00e9 inform\u00e1cie o pre\u017e\u00edvan\u00ed, resp. \u00famrt\u00ed. \u00daspe\u0161ne sme stanovili medi\u00e1n v skupin\u00e1ch: stredn\u00e1, zl\u00e1 <\/em>a ve\u013e<\/em>mi zl\u00e1<\/em>. V skupin\u00e1ch ve\u013emi dobr\u00e1 <\/em>a dobr\u00e1 <\/em>mali pacienti vy\u0161\u0161ie pre\u017e\u00edvanie a pri ich progn\u00f3zach neklesla pravdepodobnos\u0165 pre\u017eitia pod 0,5 (50 %). Nez\u00edskali sme teda relevantn\u00e9 d\u00e1ta, ke\u010f\u017ee ve\u013ek\u00e1 \u010das\u0165 pacientov e\u0161te st\u00e1le \u017eije. Grafick\u00e9 zobrazenie demon\u0161truje, \u017ee s hor\u0161ou progn\u00f3zou kles\u00e1 aj pre\u017e\u00edvanie pacientov. V tabu\u013eke 3 <\/em><\/strong>je zhrnutie \u00fadajov z jednotliv\u00fdch rizikov\u00fdch skup\u00edn. Zobrazen\u00e9 s\u00fa po\u010dty pacientov v skupin\u00e1ch, po\u010det \u00famrt\u00ed a \u017eij\u00facich v danej skupine a percento pre\u017e\u00edvania skupiny.<\/p>\n

     <\/p>\n

    3.\u00a0 Stanovenie \u00faspe\u0161nosti pou\u017eit\u00fdch met\u00f3d<\/h4>\n

    V\u0161etk\u00fdmi troma met\u00f3dami s\u00fa\u010dasne bolo vy\u0161etren\u00fdch 80 pacientov. Tento s\u00fabor sl\u00fa\u017eil na porovnanie \u00faspe\u0161nosti pou\u017eit\u00fdch met\u00f3d. Porovn\u00e1vali sme po\u010det \u00faspe\u0161n\u00fdch anal\u00fdz jednotliv\u00fdch met\u00f3d, ktor\u00e9 potvrdili aspo\u0148 jednu chroanal\u00fdzach. Uk\u00e1zala sa ako najcitlivej\u0161ia met\u00f3da na odhalenie pr\u00edtomnosti aber\u00e1ci\u00ed. Anal\u00fdza karyotypu bola pozit\u00edvna pri 60 (75 %) pr\u00edpadoch. Skr\u00edningov\u00e1 met\u00f3da MLPA preuk\u00e1zala porovnate\u013en\u00fa \u00faspe\u0161nos\u0165 ako cytogenetick\u00e1 anal\u00fdza. Zaznamenala pozit\u00edvny n\u00e1lez u 59 (74 %) pacientov.<\/p>\n

     <\/p>\n

    Diskusia<\/strong><\/p>\n

    MDS je ochorenie preva\u017ene star\u0161\u00edch \u013eud\u00ed s medi\u00e1nom 70 rokov v \u010dase diagn\u00f3zy, skupinu pod 50 rokov tvor\u00ed ~ 10 % pacientov(2). V na\u0161ej pr\u00e1ci bol vekov\u00fd medi\u00e1n 69 rokov a po\u010det pacientov mlad\u0161\u00edch ako 50 rokov bol 15 %. Incidencia MDS rap\u00eddne st\u00fapa s vekom nad 60 rokov. Greenberg a kol.(9) uv\u00e1dzaj\u00fa, \u017ee a\u017e 77 % pacientov je v \u010dase ur\u010denia diagn\u00f3zy star\u0161\u00edch ako 60 rokov. V na\u0161ej pr\u00e1ci tvorili pacienti s vekom nad 60 rokov podiel 74 %.<\/p>\n

    Na \u0161tandardn\u00e9 anal\u00fdzy bolo pou\u017eit\u00fdch 383 vzoriek kostnej drene a 237 vzoriek perif\u00e9rnej krvi. Pacienti vy\u0161etren\u00ed zo vzorky kostnej drene mali patol\u00f3gie v 138 (36 %) pr\u00edpadoch, pacienti vy\u0161etren\u00ed zo vzorky perif\u00e9rnej krvi len v 34 (14 %) pr\u00edpadoch. V literat\u00fare sa ud\u00e1va z\u00e1chyt patol\u00f3gi\u00ed v kostnej dreni pacientov v rozsahu od 37 do 55 %(9,11,12). Ostatn\u00ed pacienti mali negat\u00edvny n\u00e1lez. \u0160t\u00fadie, ktor\u00e9 by uv\u00e1dzali z\u00e1chyt abnormal\u00edt z plnej perif\u00e9rnej krvi, nie s\u00fa k dispoz\u00edcii. Na z\u00e1klade na\u0161ich v\u00fdsledkov m\u00f4\u017eeme kon\u0161tatova\u0165, \u017ee kostn\u00e1 dre\u0148 je oproti perif\u00e9rnej krvi vhodnej\u0161\u00edm biologick\u00fdm materi\u00e1lom na anal\u00fdzy. Tu samotn\u00e9 ochorenie vznik\u00e1, je tu vy\u0161\u0161\u00ed v\u00fdskyt patologick\u00fdch buniek, a t\u00fdm aj vy\u0161\u0161\u00ed z\u00e1chyt aber\u00e1ci\u00ed.<\/p>\n

    Z v\u00fdsledkov vzoriek kostnej drene 383 pacientov, ktor\u00ed boli vy\u0161etren\u00ed minim\u00e1lne dvomi met\u00f3dami z met\u00f3d cytogenetiky, FISH a MLPA, boli charakterizovan\u00e9 pr\u00edtomn\u00e9 abnormality a n\u00e1sledne bol stanoven\u00fd v\u00fdskyt t\u00fdch naj\u010dastej\u0161\u00edch. V tabu\u013eke 4 <\/em><\/strong>s\u00fa porovnan\u00e9 na\u0161e \u00fadaje v\u00fdskytu jednotliv\u00fdch patol\u00f3gi\u00ed so \u0161t\u00fadiou s dostupn\u00fdmi \u00fadajmi. Na\u0161e \u00fadaje sa s literat\u00farou vo ve\u013ekej miere zhoduj\u00fa. Tie\u017e pomery izolovan\u00fdch a kombinovan\u00fdch chromoz\u00f3mov\u00fdch zmien sa zhoduj\u00fa. Vzniknut\u00e9\u00a0rozdiely m\u00f4\u017eu by\u0165 sp\u00f4soben\u00e9 r\u00f4znou ve\u013ekos\u0165ou porovn\u00e1van\u00fdch s\u00faborov. Pou\u017eit\u00edm v\u00fdsledkov z vy\u0161etren\u00ed z\u00edskan\u00fdch met\u00f3dami cytogenetiky, FISH a MLPA bola stanoven\u00e1 progn\u00f3za pacientov. Hodnoten\u00e9 bolo zast\u00fapenie pacientov v prognostick\u00fdch skupin\u00e1ch a ich medi\u00e1n pre\u017e\u00edvania. Progn\u00f3za pacientov bola ur\u010den\u00e1 pr\u00edtomnos\u0165ou dan\u00fdch abnormal\u00edt v gen\u00f3me. Porovnan\u00edm na\u0161ich d\u00e1t so \u0161t\u00fadiami vypracovan\u00fdmi na in\u00fdch pracovisk\u00e1ch bola pozorovan\u00e1 v\u00fdznamn\u00e1 korel\u00e1cia dosiahnut\u00fdch v\u00fdsledkov. Na\u0161e v\u00fdsledky progn\u00f3z pacientov porovnan\u00e9 s in\u00fdmi \u0161t\u00fadiami s\u00fa uveden\u00e9 v tabu\u013eke 5<\/em><\/strong>. Spolu s progn\u00f3zou sme vypo\u010d\u00edtali\u00a0 medi\u00e1n\u00a0 pre\u017e\u00edvania\u00a0 351\u00a0 pacientom v jednotliv\u00fdch rizikov\u00fdch skupin\u00e1ch. V\u00fdpo\u010dtom sme z\u00edskali relevantn\u00e9 v\u00fdsledky pre skupiny: stredn\u00e1, zl\u00e1 a ve\u013emi zl\u00e1. Pre skupiny s ve\u013emi dobr\u00fdm a dobr\u00fdm rizikom sme nemali posta\u010duj\u00face \u00fadaje. N\u00e1\u0161 s\u00fabor pacientov bol sledovan\u00fd po\u010das 2 rokov a pacienti z t\u00fdchto skup\u00edn \u010dasto dosahuj\u00fa pre\u017e\u00edvanie vy\u0161\u0161ie ne\u017e 2 roky, priemerne od 5,3 do 8,7 roka(19). \u0160tatisticky sme pre tieto dve skupiny dosiahli prognostick\u00fa hodnotu, pri ktorej neklesla pravdepodobnos\u0165 pre\u017eitia pod 0,5 (50 %). Medi\u00e1n pre\u017e\u00edvania v strednej prognostickej skupine tvoril 1,6 roka. V zlej prognostickej skupine bol 1,3 roka a vo ve\u013emi zlej prognostickej skupine mal hodnotu len 0,2 roka. Na\u0161e dosiahnut\u00e9 v\u00fdsledky a porovnanie s in\u00fdmi \u0161t\u00fadiami s\u00fa zobrazen\u00e9 v tabu\u013eke 6<\/em>.<\/strong><\/p>\n

    D\u00f4vodom, pre\u010do n\u00e1m vy\u0161li ni\u017e\u0161ie hodnoty medi\u00e1nu pre\u017e\u00edvania a nedosiahli sme v\u00fdsledky pre skupiny s ve\u013emi dobrou a dobrou progn\u00f3zou, m\u00f4\u017ee by\u0165 kr\u00e1tke obdobie sledovania pacientov. Na\u0161e v\u00fdsledky potvrdili, \u017ee so zhor\u0161ovan\u00edm progn\u00f3zy sa skracuje celkov\u00e9 pre\u017e\u00edvanie pacientov.<\/p>\n

    Porovn\u00e1vali sme tie\u017e \u00faspe\u0161nos\u0165 pou\u017eit\u00fdch diagnostick\u00fdch met\u00f3d. Vyhodnocovali sme s\u00fabor pacientov, ktor\u00ed boli naraz vy\u0161etren\u00ed v\u0161etk\u00fdmi troma met\u00f3dami. Pacienti nemohli ma\u0165 negat\u00edvny n\u00e1lez a met\u00f3dy museli zachyti\u0165 aspo\u0148 jednu abnormalitu. FISH met\u00f3da sa uk\u00e1zala ako najcitlivej\u0161ia na odhalenie pr\u00edtomnosti patol\u00f3gi\u00ed. \u00daspe\u0161ne zachytila patol\u00f3giu v 85 % pr\u00edpadov. FISH m\u00e1 vysok\u00fa citlivos\u0165 a \u0161pecificitu k cie\u013eovej oblasti v DNA. Deteguje abnormality s rozl\u00ed\u0161en\u00edm od 100 kb do 1 Mb, ale zachyt\u00ed len oblas\u0165 chromoz\u00f3mu, pri ktorej je pou\u017eit\u00e1 sonda. Anal\u00fdza karyotypu (75 %) a met\u00f3da MLPA (74 %) preuk\u00e1zali porovnate\u013en\u00fa \u00faspe\u0161nos\u0165. Karyotypov\u00e1 anal\u00fdza zachyt\u00e1va numerick\u00e9 a \u0161trukt\u00farne aber\u00e1cie naraz na v\u0161etk\u00fdch chromoz\u00f3moch do detek\u010dn\u00e9ho limitu okolo 5 Mb a je obmedzen\u00e1 ne\u00faspe\u0161nou kultiv\u00e1ciou buniek v m\u00e9diu(20).<\/p>\n

    Met\u00f3da MLPA umo\u017e\u0148uje s\u00fa\u010dasne skr\u00edning 46 \u0161pecifick\u00fdch oblast\u00ed, v ktor\u00fdch s\u00fa zahrnut\u00e9 aj menej frekventovan\u00e9 MDS abnormality. Jej obmedzen\u00edm je v\u0161ak to, \u017ee nedok\u00e1\u017ee zachyti\u0165 balansovan\u00e9 genetick\u00e9 prestavby a potrebuje aspo\u0148 25 \u2013 30 % postihnut\u00fdch buniek vo vzorke(21).<\/p>\n

     <\/p>\n

    Z\u00e1ver<\/h3>\n

    Incidencia MDS v s\u00fa\u010dasnosti v Eur\u00f3pe st\u00fapa a predpoklad\u00e1 sa ka\u017edoro\u010dne n\u00e1rast o 25 000 nov\u00fdch pr\u00edpadov. D\u00f4vodom je nielen starnutie popul\u00e1cie, ale aj n\u00e1rast zne\u010distenia prostredia. Preto je ve\u013emi d\u00f4le\u017eit\u00e1 presn\u00e1 diagnostika pacientov, spr\u00e1vne stanovenie progn\u00f3zy a n\u00e1sledne nastavenie vhodnej terapie. V diagnostike aber\u00e1ci\u00ed sa dnes vyu\u017e\u00edvaj\u00fa \u0161tandardn\u00e9 genetick\u00e9 met\u00f3dy ako cytogenetick\u00e9 vy\u0161etrenie karyotypu, met\u00f3da FISH a MLPA anal\u00fdza, z ktor\u00fdch m\u00e1 ka\u017ed\u00e1 svoje nezamenite\u013en\u00e9 miesto a ich vz\u00e1jomn\u00e1 kombin\u00e1cia je z\u00e1rukou zachytenia \u010do najv\u00e4\u010d\u0161ieho po\u010dtu aber\u00e1ci\u00ed v gen\u00f3me pacienta s MDS.<\/p>\n

     <\/p>\n

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

      \n
    1. Vondr\u00e1kov\u00e1 Myelodysplastick\u00fd syndrom, diagnostika a l\u00e9\u010dba. Interni Med 2010; 12(11): 535-539.<\/li>\n
    2. Germing U, Kobbe G, Haas R, Gattermann Myelodysplastic syndromes: diagnosis, prognosis and treatment. Dtsch Arztebl Int 2013; 110(46): 783-790. doi: 10.3238\/arztebl.2013.0783.<\/li>\n
    3. Rau ATK, Shreedhara AK, Kumar Myelodysplastic Syndromes in Children: Where Are We Today? Ochsner J. 2012; 12(3): 216-220.<\/li>\n
    4. Neuwirtov\u00e1 R. Myelodysplastick\u00fd syndrom: onkohematologick\u00e9 onemocn\u011bn\u00ed vy\u0161\u0161\u00edho v\u011bku. \u010ces Ger Rev 2005; 3(2): 21-28.<\/li>\n
    5. Ad\u00e8s L, Itzykson R, Fenaux Myelodysplastic syndromes. Lancet 2014; 383(9936): 2239-2252.<\/li>\n
    6. Larson RA. Therapy-related myeloid neoplasms. Haematologica 2009; 94(4): 454-459.<\/li>\n
    7. Haase D, Germing U, Schanz J, et New insights into the prognostic impact of the karyotype in MDS and correlation with subtypes: evidence from a core dataset of 2124 patients. Blood 2007; 110(13): 4385-4395.<\/li>\n
    8. Jonas BA, Greenberg MDS prognostic scoring systems \u2013 past, present, and future. Best Pract Res Clin Haematol 2015; 28(1): 3-13.<\/li>\n
    9. Greenberg PL, Tuechler H, Schanz J, et Revised international prognostic scoring system for myelodysplastic syndromes. Blood 2012; 120(12): 2454-65.<\/li>\n
    10. Zatkova A, Merk S, Wendehack M, et AML\/MDS with 11q\/MLL Amplification Show Characteristic Gene Expression Signature and Interplay of DNA Copy Number Changes. GENES, CHROMOSOMES & CANCER 2009; 48: 510-520.<\/li>\n
    11. Haase Cytogenetic features in myelodysplastic syndromes. Ann Hematol 2008; 87(7): 515-526.<\/li>\n
    12. Schanz J, T\u00fcchler H, Sol\u00e9 F, et New comprehensive cytogenetic scoring system for primary myelodysplastic syndromes (MDS) and oligoblastic acute myeloid leukemia after MDS derived from an international database merge. J Clin Oncol 2012; 30(8): 820-829.<\/li>\n
    13. Schouten JP, McElgunn CJ, Waaijer R, et Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res 2002; 30(12): e57.<\/li>\n
    14. Donahue AC, Abdool AK, Gaur R, et Multiplex ligation-dependent probe amplification for detection of chromosomal abnormalities in myelodysplastic syndrome and acute myeloid leukemia. Leuk Res 2011; 35(11): 1477-83.<\/li>\n
    15. Sidney LE, Branch MJ, Dunphy SE, et Concise review: evidence for CD34 as a common marker for diverse progenitors. Stem Cells 2014; 32(6): 1380-89.<\/li>\n
    16. Marisavljevic D, Kraguljac-Kurtovic Biological implications of circulating CD34(+) cells in myelodysplastic syndromes. J BUON 2010; 15(4): 753-757.<\/li>\n
    17. Liang X, Xu K, Xu J, et Preparation of immunomagnetic nanoparticles and their application in the separation of mouse CD34+ hematopoietic stem cells. Journal of Magnetism and Magnetic Materials 2009; 321(12): 1885-88.<\/li>\n
    18. Demirer GS, Okur AC, Kizilel Synthesis and design of biologically inspired biocompatible iron oxide nanoparticles for biomedical applications. J Mater Chem B 2015; 3: 7831-7849.<\/li>\n
    19. https:\/\/www.mds-foundation.org\/wp-content\/uploads\/2<\/a>011\/12\/2-Revised-<\/a>pdf<\/li>\n
    20. https:\/\/w<\/a>wbiomnis.com\/wpcontent\/uploads\/2016\/04\/56-INTGB-Focus_<\/a> Karyotyping_SNP_array.pdf<\/li>\n
    21. http:\/\/www.mlpa.com\/WebForms\/WebFormDBData.aspx?Tag=_G1U3P-<\/a> YAOzf2SFuaxkiqa4YjruAIwx3T3q8uAJJ_V-Ws<\/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 Myelodysplastick\u00fd syndr\u00f3m (MDS) predstavuje heterog\u00e9nnu skupinu chronick\u00fdch myeloidn\u00fdch ochoren\u00ed s p\u00f4vodom v kostnej dreni. Ide o klon\u00e1lnu poruchu krvotvorby ako d\u00f4sledok mut\u00e1ci\u00ed vznikaj\u00facich v pluripotentn\u00fdch kme\u0148ov\u00fdch bunk\u00e1ch. Vplyvom<\/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":[963,341,964,962,961],"class_list":["post-1533","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-chromosomal-aberrations","tag-chromozomove-aberacie-en","tag-diagnostics","tag-myelodysplastic-syndrome","tag-myelodysplasticky-syndrom-en","typ_clanku-original-work"],"acf":{"abstrakt":"

      The myelodysplastic syndrome (MDS) is a heterogeneous group of clonal haematopoietic disorders originated in the bone marrow. It is characterized by ineffective haematopoiesis, dysplastic changes in blood cells and cytopenia in peripheral blood. Patients are mostly older, and around 30% of all patients later transform to acute myeloid leukemia (AML). One of the essential parameters in diagnosis is an evidence of chromosomal aberrations characteristic for this disease. Chromosomal changes are detected in approximately 50% of patients with primary MDS, and more than 80% patients are detected with secondary MDS. According to specific chromosomal abnormalities, it is possible to predict a patient\u2019s prognosis, his overall survival and an appropriate treatment. We detected these abnormalities by cytogenetic method using conventional banding techniques, fluorescence in situ hybridization (FISH) and multiplex ligation dependent probe amplification (MLPA).<\/p>\n

      Keywords:<\/strong> myelodysplastic syndrome, chromosomal aberrations, diagnostics<\/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
      • Genetically important aberrations in patients with mye-lodysplastic syndrome and laboratory methods of their detection<\/li>\r\n \t
      • Molecular \u2013 genetic diagnostics of Human Papillomavirus (HPV) and monitoring of HPV patients<\/li>\r\n \t
      • Laboratory diagnostic possibilities for Clostridium difficile infections<\/li>\r\n \t
      • Chorangiosis of Placenta - Disorder of Unclear Etiology (Case Report and Overview of Current Knowledge)<\/li>\r\n \t
      • 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":"65","upload_clanok":{"ID":1518,"id":1518,"title":"Geneticky v\u00fdznamn\u00e9 aber\u00e1cie u pacientov s myelodysplastick\u00fdm","filename":"Geneticky-v\u00fdznamn\u00e9-aber\u00e1cie-u-pacientov-s-myelodysplastick\u00fdm.pdf","filesize":607445,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2018\/11\/Geneticky-v\u00fdznamn\u00e9-aber\u00e1cie-u-pacientov-s-myelodysplastick\u00fdm.pdf","link":"https:\/\/www.newslab.sk\/en\/geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-syndromom-a-sposoby-ich-detekcie\/geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-2\/","alt":"","author":"7","description":"","caption":"","name":"geneticky-vyznamne-aberacie-u-pacientov-s-myelodysplastickym-2","status":"inherit","uploaded_to":1533,"date":"2018-11-09 08:52:48","modified":"2018-11-09 08:52:48","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\/1533","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=1533"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1533\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1533"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1533"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1533"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}