{"id":2408,"date":"2022-02-09T14:49:40","date_gmt":"2022-02-09T13:49:40","guid":{"rendered":"https:\/\/www.newslab.sk\/vyuzitie-hmotnostnej-spektrometrie-v-diagnostike-poruch-glykozylacie\/"},"modified":"2022-02-10T02:20:04","modified_gmt":"2022-02-10T01:20:04","slug":"the-application-of-mass-spectrometry-in-diagnostics-of-glycosylation-disorders","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/the-application-of-mass-spectrometry-in-diagnostics-of-glycosylation-disorders\/","title":{"rendered":"The application of mass spectrometry in diagnostics of glycosylation disorders"},"content":{"rendered":"

*A rare case of autochthonous human dirofilariasis with the manifestation of pseudotumor of the epididymis caused by helminth Dirofilaria repens<\/strong><\/span><\/p>\n

 <\/p>\n

\u00davod<\/h3>\n

Glykozyl\u00e1cia je esenci\u00e1lna modifik\u00e1cia prote\u00ednov, ktor\u00e1 z\u00e1sadne ovplyv\u0148uje ich funkciu, stabilitu, aktivitu a mnoh\u00e9 in\u00e9 vlastnosti. Na rozdiel od transkripcie a transl\u00e1cie je to netempl\u00e1tov\u00fd proces, katalyzovan\u00fd stovkami r\u00f4znych enz\u00fdmov a transport\u00e9rov, prebiehaj\u00faci v endoplazmatickom retikule, Golgiho apar\u00e1te a cytozole. Dnes je zn\u00e1me, \u017ee porucha homeost\u00e1zy organizmu m\u00f4\u017ee negat\u00edvne ovplyvni\u0165 s\u00fahru t\u00fdchto glykozyla\u010dn\u00fdch metabolick\u00fdch kask\u00e1d. Aberantn\u00e1 glykozyl\u00e1cia bola pozorovan\u00e1 pri r\u00f4znych onkologick\u00fdch, neurodegenerat\u00edvnych, autoimunitn\u00fdch a in\u00fdch ochoreniach(1-3) pravdepodobne ako sekund\u00e1rny jav; ale takisto pri kongenit\u00e1lnych poruch\u00e1ch glykozyl\u00e1cie (CDG), ktor\u00e9 s\u00fa prim\u00e1rne sp\u00f4soben\u00e9 mut\u00e1ciou g\u00e9nov komplexnej glykozyla\u010dnej dr\u00e1hy.
\nP\u00f4vodne sa skupina CDG delila na dva typy (typ I a II) pod\u013ea profilu IEF transfer\u00ednu (Tf), pri\u010dom do skupiny CDG I patrili defekty sp\u00f4soben\u00e9 pri synt\u00e9ze N-glyk\u00e1nov vzniknut\u00e9 v cytozole alebo v endoplazmatickom retikule a do CDG II defekty vzniknut\u00e9 v Golgiho apar\u00e1te pri \u00faprave glyk\u00e1nov. Toto rozde-lenie zah\u0155\u0148alo len poruchy N-glykozyl\u00e1cie. Vzh\u013eadom na narastaj\u00faci po\u010det ochoren\u00ed a neust\u00e1le odha\u013eovanie nov\u00fdch por\u00fach v dr\u00e1hach glykozyl\u00e1cie bolo nutn\u00e9 po op\u00edsan\u00ed viac ako 40 ochoren\u00ed v roku 2009 klasifik\u00e1ciu aj nomenklat\u00faru zmeni\u0165. Na ozna\u010denie ochorenia sa v s\u00fa\u010dasnosti pou\u017e\u00edva n\u00e1zov mutova-n\u00e9ho g\u00e9nu spolu so skratkou CDG. V s\u00fa\u010dasnosti je zn\u00e1mych viac ako 160 r\u00f4znych por\u00fach glykozyl\u00e1cie. Nov\u00e1 klasifik\u00e1cia rozde\u013euje cel\u00fa skupinu na: (a) poruchy N-glykozyl\u00e1cie prote\u00ed-nov, (b) poruchy O-glykozyl\u00e1cie prote\u00ednov, (c) poruchy synt\u00e9zy glykozylfosfatidylinozitolovej kotvy a glykolipidov, (d) poruchy viacer\u00fdch dr\u00e1h glykozyl\u00e1cie a poruchy ostatn\u00fdch dr\u00e1h.
\nBiochemick\u00fdm skr\u00edningom N-glykozyla\u010dn\u00fdch, O-glykozyla\u010dn\u00fdch a zmie\u0161an\u00fdch por\u00fach a por\u00fach synt\u00e9zy dolicholu je IEF Tf, apolipoprote\u00ednu C-III zo s\u00e9ra a vy\u0161etrenie profilu dolicholov v mo\u010di.
\nKlinick\u00e9 pr\u00edznaky CDG \u010dasto zah\u0155\u0148aj\u00fa multiorg\u00e1nov\u00e9 postihnutie. Medzi \u010dast\u00fdmi n\u00e1lezmi s\u00fa polystigmatiz\u00e1cia, poruchy rastu, psychomotorick\u00e1 retard\u00e1cia, poruchy imunity, hypot\u00f3nia, z\u00e1chvaty a muskuloskeletov\u00e9 abnormality. Klinick\u00e9 manifest\u00e1cie takmer u v\u0161etk\u00fdch CDG pacientov sa objavuj\u00fa u\u017e v doj\u010denskom veku(4). Biochemick\u00e9 parametre zah\u0155\u0148aj\u00fa koagulopatie a zv\u00fd\u0161en\u00e9 hladiny pe\u010de\u0148ov\u00fdch transamin\u00e1z. V\u00a0 pr\u00edpade PMM2-CDG, subtypu s\u00a0 celosvetovo najvy\u0161\u0161ou frekvenciou v\u00fdskytu heterogenita klinick\u00fdch prejavov zah\u0155\u0148a skor\u00fa neonat\u00e1lnu mortalitu a\u017e po dosiahnutie norm\u00e1lnej do-spelosti(5). Korel\u00e1cie medzi genotypom a fenotypom s\u00fa u jednotliv\u00fdch pacientov ve\u013emi variabiln\u00e9, \u010do spolu s ne\u0161pecifick\u00fdmi klinick\u00fdmi pr\u00edznakmi zara\u010fuje tieto ochorenia medzi \u0165a\u017eko diagnostikovate\u013en\u00e9.<\/p>\n

Materi\u00e1l a\u00a0met\u00f3da<\/h3>\n

V roku 2012 bol v Centre dedi\u010dn\u00fdch metabolick\u00fdch por\u00fach (CDMP) N\u00daDCH zaveden\u00fd selekt\u00edvny skr\u00edning CDG zalo\u017een\u00fd na met\u00f3de izoelektrickej fokus\u00e1cie (IEF) s\u00e9rov\u00e9ho transfer\u00ednu. Za obdobie rokov 2012 \u2013 2020 bolo celkovo analyzo van\u00fdch 3\u2009600 vzoriek pacientov so suspektn\u00fdm podozren\u00edm na CDG odoslan\u00fdch z metabolick\u00fdch ambulanci\u00ed a klinick\u00fdch pracov\u00edsk v r\u00e1mci cel\u00e9ho Slovenska. Princ\u00edp IEF Tf spo\u010d\u00edva v separ\u00e1cii izoforiem transfer\u00ednu (biomarkera nes\u00faceho dve N-glykozyla\u010dn\u00e9 miesta) na z\u00e1klade ich izoelektrick\u00e9ho bodu s n\u00e1slednou imunodetekciou a denzitometrickou kvantifik\u00e1ciou. Met\u00f3da umo\u017e\u0148uje rozl\u00ed\u0161i\u0165 poruchu glykozyl\u00e1cie v synt\u00e9ze oligosacharidick\u00e9ho prekurzora v endoplazmatickom retikule (CDG typ I) od poruchy v procesovan\u00ed glyk\u00e1nov (CDG typ II), ktor\u00e9 prebieha v Golgiho apar\u00e1te. T\u00e1to met\u00f3da je ve\u013e-mi spo\u013eahliv\u00e1, reprodukovate\u013en\u00e1 a predstavuje celosvetovo tzv. zlat\u00fd \u0161tandard pre CDG diagnostiku, nedok\u00e1\u017ee v\u0161ak ur\u010di\u0165 konkr\u00e9tne miesto preru\u0161enia glykozyla\u010dnej metabolickej dr\u00e1hy. Odhaduje sa, \u017ee a\u017e 40 % zachyten\u00fdch CDG pacientov st\u00e1le nem\u00e1 potvrden\u00fa presn\u00fa molekul\u00e1rnu podstatu(6). Diagnostika t\u00fdchto zriedkav\u00fdch ochoren\u00ed vy\u017eaduje personalizovan\u00fd pr\u00edstup, kde nach\u00e1dza svoje uplatnenie spolupr\u00e1ca klinick\u00fdch diagnostikov s vedeck\u00fdmi laborat\u00f3riami.
\nChemick\u00fd \u00fastav Slovenskej akad\u00e9mie vied sa v\u010faka boha-t\u00fdm sk\u00fasenostiam v oblasti \u0161trukt\u00farnej anal\u00fdzy glykokonjug\u00e1tov zapojil do region\u00e1lnej (CDMP N\u00daDCH) i medzin\u00e1rodnej spolupr\u00e1ce (v r\u00e1mci CDG\u2008&\u2008ALLIES \u2013 PPAIN siete). Zaveden\u00fd \u0161tandardn\u00fd opera\u010dn\u00fd protokol anal\u00fdzy N-glykoprofilu vyu\u017e\u00edva 10 mikrolitrov krvn\u00e9ho s\u00e9ra alebo pribli\u017ene 100 mikrogramov imunoafinitne izolovan\u00e9ho glykobiomarkera (napr. Tf). Glyk\u00e1ny s\u00fa enzymaticky od\u0161tiepen\u00e9 a separovan\u00e9 od prote\u00ednov prostredn\u00edctvom \u201esolidphase extrakcie\u201c. S cie\u013eom zv\u00fd\u0161enia intenz\u00edt sign\u00e1lov v n\u00e1sledne nameran\u00fdch MALDI TOF hmotnostn\u00fdch spektr\u00e1ch (MS) sa vzorky derivatizuj\u00fa permetyl\u00e1ciou. V pr\u00edpade potreby kvantifik\u00e1cie jednotliv\u00fdch \u0161trukt\u00far sa vyu\u017e\u00edva vysoko\u00fa\u010dinn\u00e1 kvapalinov\u00e1 chromatografia (HPLC) v\u00a0 kombin\u00e1cii s\u00a0 MS. Reprezentat\u00edvny glykoprofil Tf z\u00edskan\u00fd zo vzorky negat\u00edvnej kontroly met\u00f3dami tradi\u010dn\u00e9ho selekt\u00edvneho skr\u00edningu, HPLC a MS je zn\u00e1zornen\u00fd na obr\u00e1zku 1. Pri podozren\u00ed na poruchu glykozyl\u00e1cie prv\u00e9ho typu (CDG I) je vhodn\u00e1 anal\u00fdza neutr\u00e1lnej frakcie N-glyk\u00e1nov prostredn\u00edctvom MALDI TOF MS a pri podozren\u00ed na CDG II je, naopak, naj-vhodnej\u0161\u00edm pr\u00edstupom anal\u00fdza sialylovan\u00fdch glyk\u00e1nov, kde mo\u017eno pozorova\u0165 pr\u00edpadn\u00e9 defekty v kone\u010dnom procesovan\u00ed glyk\u00e1nov u\u017e naviazan\u00fdch na vznikaj\u00faci prote\u00edn.<\/p>\n

V\u00fdsledky a\u00a0diskusia<\/h3>\n

V\u00a0 predlo\u017eenej pr\u00e1ci prezentujeme dvoch vybran\u00fdch slo-vensk\u00fdch pacientov, ktor\u00fdm bola diagnostikovan\u00e1 CDG s nasleduj\u00facou skr\u00e1tenou epikr\u00edzou:
\nPacient I\u00a0 \u2013 novorodenec, respira\u010dn\u00e1 insuficiencia, hy-poglyk\u00e9mia, trombocytop\u00e9nia, hypokolagua\u010dn\u00fd stav, skele-tov\u00e9 abnormality, nefropatia, hepatopatia, opakuj\u00face sa res-pira\u010dn\u00e9 infekcie. IEF Tf pouk\u00e1zala na CDG typu I. Pacient exitoval vo veku 164 dn\u00ed. Sekvenovan\u00edm klinick\u00e9ho ex\u00f3mu bola potvrden\u00e1 homozygotn\u00e1 mut\u00e1cia v g\u00e9ne ALG12, k\u00f3duj\u00faceho esenci\u00e1lny enz\u00fdm z\u00fa\u010dast\u0148uj\u00faci sa na v\u00fdstavbe oligo-sacharidick\u00e9ho prekurzora v endoplazmatickom retikule(7).
\nPacient II\u00a0 \u2013 vek v\u00a0 \u010dase stanovenia diagn\u00f3zy 12 rokov; k dne\u0161n\u00e9mu d\u0148u v dobrom klinickom stave; pretrv\u00e1vaj\u00faci hypokoagula\u010dn\u00fd stav, mierne zv\u00fd\u0161en\u00e9 hladiny AST, ALT, hypocholesterol\u00e9mia, hypobetalipoprotein\u00e9mia. IEF Tf pouk\u00e1za-la na CDG typu II. Na z\u00e1klade ex\u00f3mov\u00e9ho sekvenovania bola u pacienta potvrden\u00e1 de novo mut\u00e1cia v g\u00e9ne SLC37A4, zod-povedn\u00e1 za mislokaliz\u00e1ciu gluk\u00f3za-6-fosf\u00e1tov\u00e9ho transport\u00e9ra(8).<\/p>\n

U prezentovan\u00fdch CDG I aj II pacientov bola na z\u00e1klade pozit\u00edvneho v\u00fdsledku zo selekt\u00edvneho skr\u00edningu vykonan\u00e1 anal\u00fdza s\u00e9rov\u00fdch N-glyk\u00e1nov prostredn\u00edctvom MALDI TOF MS. V pr\u00edpade pacienta I boli pozorovan\u00e9 zn\u00ed\u017een\u00e9 hladiny GlcNAc-2Man8-9 (m\/z 2192.2 a 2396.3). Hladina GlcNAc2Man9Glc1 (m\/z 2600.6) bola u pacienta I nedetegovate\u013en\u00e1. V pr\u00edpade pacienta\u00a0 II boli pozorovan\u00e9 signifikantne zv\u00fd\u0161en\u00e9 hladiny \u201ehybridn\u00fdch\u201c \u0161trukt\u00far Hex6GlcNAc3NeuAc1 (m\/z\u00a0 2390.2), Hex5GlcNAc3NeuAc1 (m\/z\u00a0 2186.1), a\u00a0 Hex4GlcNAc3Neu-Ac1 (m\/z 1982.0). Reprezentat\u00edvne spektr\u00e1 porovnan\u00e9 s negat\u00edvnymi kontrolami, napovedaj\u00face o\u00a0 prim\u00e1rnej poruche v glykozyl\u00e1cii, s\u00fa zn\u00e1zornen\u00e9 na obr\u00e1zku 2. (GlcNAc \u2013 N-ace-tylglukozam\u00edn, Man \u2013 man\u00f3za, Glc \u2013 gluk\u00f3za, NeuAc \u2013 kys. sialov\u00e1).
\nU pacienta I signifikantn\u00fd pokles hlad\u00edn GlcNAc2Man8-9 vo frakcii neutr\u00e1lnych s\u00e9rov\u00fdch N-glyk\u00e1nov pouk\u00e1zal na zn\u00ed\u017een\u00fa aktivitu \u03b1-manozyltransfer\u00e1zy 8, k\u00f3dovanej g\u00e9nom ALG12. Tento enz\u00fdm katalyzuje pridanie \u00f4smeho man\u00f3zov\u00e9 ho zvy\u0161ku na vznikaj\u00faci oligosacharid, viazan\u00fd prostredn\u00edc-tvom lipidov\u00e9ho nosi\u010da na membr\u00e1nu endoplazmatick\u00e9ho retikula. Anal\u00fdza klinick\u00e9ho ex\u00f3mu prostredn\u00edctvom Clinical Exome Solution panelu sekvenovania novej gener\u00e1cie (z\u00a0an-gl. \u201enext generation sequencing\u201c, NGS) identifikovala homo-zygotn\u00fd variant c.1439T>C p.(Leu480Pro) v\u00a0ex\u00f3ne 10 g\u00e9nu ALG12. K\u00a0dne\u0161n\u00e9mu d\u0148u bolo celosvetovo identifikovan\u00fdch iba 16 pacientov s\u00a0t\u00fdmto ochoren\u00edm.
\nPoruchy glykozyl\u00e1cie druh\u00e9ho typu s\u00fa ve\u013ekou v\u00fdzvou pre diagnostiku, preto\u017ee aberantn\u00fd glykoprofil \u010dasto nepoukazuje na konkr\u00e9tny defekt v glykozyla\u010dnej kask\u00e1de. Prezentovan\u00e9 v\u00fdsledky z anal\u00fdzy s\u00e9rov\u00fdch N-glyk\u00e1nov pacienta II, ktor\u00e9ho molekul\u00e1rna podstata bola objasnen\u00e1 na zahrani\u010d-nom pracovisku(8), v\u0161ak na z\u00e1klade akumul\u00e1cie aberantn\u00fdch hybridn\u00fdch \u0161trukt\u00far poukazuj\u00fa na prim\u00e1rnu poruchu v glykozyl\u00e1cii. NGS anal\u00fdza ex\u00f3mu pouk\u00e1zala na de novo mut\u00e1ciu c.1267C > T (R423*) v g\u00e9ne SLC37A4 k\u00f3duj\u00facom transport\u00e9r gluk\u00f3zo6fosf\u00e1tu(8). Na z\u00e1klade literat\u00fary strata funkcie trans-port\u00e9ra gluk\u00f3zo-6-fosf\u00e1tu vedie k lyzoz\u00f3movej poruche s n\u00e1zvom \u201eGlycogen storage disorder (GSD)\u201c typu Ib(9), ale \u0161pecifick\u00e1 mut\u00e1cia v tomto g\u00e9ne m\u00f4\u017ee vies\u0165 aj k intracelul\u00e1rnej mislokaliz\u00e1cii tohto transport\u00e9ra s \u00faplne odli\u0161nou klinickou manifest\u00e1ciou(8).
\nHmotnostn\u00e1 spektrometria predstavuje v\u00fdznamn\u00fd pr\u00ednos v diagnostike CDG prv\u00e9ho aj druh\u00e9ho typu a m\u00f4\u017ee by\u0165 \u0161tandardne zaveden\u00e1 ako nasleduj\u00faci krok, nadv\u00e4zuj\u00faci na pozit\u00edvny, respekt\u00edve hrani\u010dn\u00fd v\u00fdsledok selekt\u00edvneho skr\u00edningu. \u0160trukt\u00farna anal\u00fdza predstavuje esenci\u00e1lnu s\u00fa\u010das\u0165 personalizovanej diagnostiky a aj v\u010faka svojej ni\u017e\u0161ej finan\u010dnej a \u010dasovej n\u00e1ro\u010dnosti by mala predch\u00e1dza\u0165 molekul\u00e1rnogenetick\u00fdm anal\u00fdzam (NGS). Odhaduje sa v\u0161ak, \u017ee pre technologick\u00fa n\u00e1-ro\u010dnos\u0165 personalizovanej diagnostiky a \u0161irok\u00fa heterogenitu klinick\u00fdch pr\u00edznakov celosvetovo st\u00e1le ost\u00e1va mnoho CDG pacientov poddiagnostikovan\u00fdch. Zv\u00fd\u0161enie povedomia laickej i odbornej verejnosti o t\u00fdchto ochoreniach a spolupr\u00e1ca klinick\u00fdch a vedeck\u00fdch pracov\u00edsk v\u0161ak m\u00f4\u017ee v\u00fdznamne prispie\u0165 k v\u010dasn\u00e9mu a spo\u013eahliv\u00e9mu ur\u010deniu diagn\u00f3zy, \u010do m\u00f4\u017ee ma\u0165 aj v\u00fdrazn\u00fd vplyv na kvalitu \u017eivota pacientov a ich rod\u00edn.<\/p>\n

Z\u00e1ver<\/h3>\n

V diagnostike zriedkav\u00fdch ochoren\u00ed nach\u00e1dza spolupr\u00e1ca medzi vedeck\u00fdmi in\u0161tit\u00faciami a diagnostick\u00fdmi pracoviskami ve\u013ek\u00e9 uplatnenie. Pokro\u010dil\u00e9 analytick\u00e9 met\u00f3dy ako hmotnostn\u00e1 spektrometria a kvapalinov\u00e1 chromatografia m\u00f4\u017eu by\u0165 napriek svojej technickej n\u00e1ro\u010dnosti optimalizovan\u00e9 a automatizovan\u00e9 do vysokopriepustn\u00fdch form\u00e1tov a roz\u0161\u00edri\u0165 tak mo\u017enosti tzv. personalizovanej diagnostiky s individu\u00e1lnym pr\u00edstupom ku ka\u017ed\u00e9mu pacientovi. Pr\u00e1ve d\u00f4kladn\u00e1 korel\u00e1cia v\u00fdsledkov \u0161trukt\u00farnej anal\u00fdzy biomarkerov a klinick\u00fdch, biochemick\u00fdch a genetick\u00fdch vy\u0161etren\u00ed m\u00f4\u017ee pom\u00f4c\u0165 objasni\u0165 molekul\u00e1rnu podstatu t\u00fdchto raritn\u00fdch ochoren\u00ed.<\/p>\n

Konflikt z\u00e1ujmov<\/h3>\n

Autori deklaruj\u00fa, \u017ee nemaj\u00fa \u017eiadny konflikt z\u00e1ujmov a z\u00e1rove\u0148 \u010destne vyhlasuj\u00fa, \u017ee pr\u00e1ca nebola publikovan\u00e1 v inom periodiku.<\/p>\n

Po\u010fakovanie<\/strong><\/h3>\n

T\u00e1to pr\u00e1ca bola podporen\u00e1 Ministerstvom zdravotn\u00edctva SR v\u00a0 r\u00e1mci projektu s\u00a0 registra\u010dn\u00fdm \u010d\u00edslom 2019\/7-CH\u00daSAV-4, grantom \u010d. 2\/0060\/21 Slovenskej grantovej agent\u00fary pre vedu VEGA. T\u00e1to publik\u00e1cia vznikla v\u010faka podpore v r\u00e1mci Opera\u010dn\u00e9ho programu Integrovan\u00e1 \u0161trukt\u00fara pre projekt: \u0160t\u00fadium \u0161trukt\u00farnych zmien komplexn\u00fdch glykokonjug\u00e1tov v\u00a0 procese dedi\u010dn\u00fdch metabolick\u00fdch a civiliza\u010dn\u00fdch ochoren\u00ed, ITMS: 313021Y920, spolufinancovan\u00fd zo zdrojov Eur\u00f3pskeho fondu region\u00e1lneho rozvoja a MZ \u010cR-RVO-VFN64165 Autori tohto \u010dl\u00e1nku s\u00fa \u010dlenmi Eur\u00f3pskej referen\u010dnej siete pre zriedkav\u00e9 metabolick\u00e9 poruchy (European Reference Network for Rare Hereditary Metabolic Disorders, MetabERN \u2013 Project ID No 739543).<\/p>\n

 <\/p>\n

LITERAT\u00daRA<\/h3>\n

1. Moffett S, Shiao TC, Mousavifar L, et al. Aberrant glycosylation patterns on cancer cells: Therapeutic opportunities for glycodendrimers\/metallodendrimers oncology. WIREs Nanomedicine and Nanobiotechnology 2021;13(1):e1659.<\/p>\n

2. Shafi S, Singh A, Gupta P, et al. Deciphering the Role of Aberrant Protein Post-Translational Modification in the Pathology of Neurodegeneration. CNS Neurol Disord \u2013 Drug Targets 2021;20(1):54\u201367.<\/p>\n

3. Kennedy PGE, Graner M, Pointon T, et al. Aberrant Immunoglobulin G Glycosylation in Multiple Sclerosis J Neuroimmune Pharmacol. Publikovan\u00e9 online 03. m\u00e1j 2021. doi:10.1007\/s11481-021-09996-1<\/p>\n

4. Lekka DE, Brucknerova J, Salingova A, et al. Congenital disorders of glycosylation \u2013 an umbrella term for rapidly expanding group of rare genetic metabolic disorders \u2013 importance of physical investigation. Bratislava Med J 2021;122(03):190\u2013195.<\/p>\n

5. Morena\u2010Barrio ME, Sabater M, Morena\u2010Barrio B, et al. ALG12\u2010CDG: An unusual patient without intellectual disability and facial dysmorphism, and with a\u00a0novel variant. Mol Genet Genomic Med 2020;8(8):e1304.<\/p>\n

6. Jones MA, Bhide S, Chin E, et al. Targeted polymerase chain reaction-based enrichment and next generation sequencing for diagnostic testing of congenital disorders of glycosylation. Genet Med 2011;13(11):921\u2013932.<\/p>\n

7. Ziburov\u00e1 J, Nem\u010dovi\u010d M, \u0160est\u00e1k S, et al. A novel homozygous mutation in the human ALG12 gene results in an aberrant profile of oligomannose N-glycans in patient\u2019s\u00a0serum. Am J Med Genet Part A\u00a02021;185(11):3494\u20133501.<\/p>\n

8. Marquardt T, Bzduch V, Hogrebe M, et al. SLC37A4-CDG: Mislocal-ization of the glucose-6-phosphate transporter to the Golgi causes a\u00a0 new congenital disorder of glycosylation. Mol Genet Metab Reports 2020;25:100636.<\/p>\n

9. Chou JY, Jun HS, Mansfield BC. Type I glycogen storage diseases: disorders of the glucose-6-phosphatase\/glucose-6-phosphate transporter complexes. J Inherit Metab Dis 2015;38(3):511\u2013519.<\/p>\n

 <\/p>\n","protected":false},"excerpt":{"rendered":"

*A rare case of autochthonous human dirofilariasis with the manifestation of pseudotumor of the epididymis caused by helminth Dirofilaria repens   \u00davod Glykozyl\u00e1cia je esenci\u00e1lna modifik\u00e1cia prote\u00ednov, ktor\u00e1 z\u00e1sadne ovplyv\u0148uje ich funkciu, stabilitu, aktivitu a mnoh\u00e9 in\u00e9 vlastnosti. Na rozdiel od transkripcie a transl\u00e1cie je to netempl\u00e1tov\u00fd proces, katalyzovan\u00fd stovkami r\u00f4znych enz\u00fdmov a transport\u00e9rov, prebiehaj\u00faci<\/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":[649,964,1478],"class_list":["post-2408","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-congenital-disorders-of-glycosylation","tag-diagnostics","tag-mass-spectrometry","typ_clanku-original-work"],"acf":{"abstrakt":"

Congenital disorders of glycosylation (CDG) are rare hereditary diseases caused by mutations of genes coding enzymes of complex glycosylation pathways. Common clinical manifestation includes growth and psychomotor retardation, immunodeficiency, hypotonia, seizures, or musculoskeletal abnormalities. Because of the heterogeneity of the clinical spectrum, CDG is difficult to diagnose. Isoelectric focusing of transferrin (IEF) with immunodetection enables distinguishing between two primary N-glycosylation disorders (type I and II). In contrast, isoelectric focusing of apolipoprotein C-III is used to diagnose O-glycosylation disorders, as this glycobiomarker bears one O-glycosylation site. For closer specification of subtype upon positive or borderline IEF result, MALDITOF mass spectrometry, which enables to focus the diagnostic process on the causal gene, mostly in unclear cases, can be utilized. In this study, we present two patients diagnosed with different types of N-glycosylation disorders. The patient I possessed a mutation in gene ALG12, coding \u03b1mannosyltransferase 8. In this patient\u2019s serum, significantly decreased levels of its products were observed. In the case of the patient II, mutation of the SLC37A4 gene caused the misallocation of the glucose-6-phosphate transporter. Analysis of this patient’s glycoprofile revealed increased levels of hybrid glycans. Diagnostics of CDG by MALDITOF mass spectrometry enables the personalization of treatment and helps clarify the molecular basis of these disorders.<\/p>\n

Keywords:<\/strong> diagnostics, congenital disorders of glycosylation, mass spectrometry<\/p>\n","casopis":[{"ID":2417,"post_author":"7","post_date":"2022-02-08 13:25:20","post_date_gmt":"2022-02-08 12:25:20","post_content":"Druh\u00e9 vydanie \u010dasopisu laborat\u00f3rnej medic\u00edny 2021\/2<\/strong>\r\n

    \r\n \t
  • Molekul\u00e1rny mechanizmus karcinogen\u00e9zy indukovanej prostredn\u00edctvom bakt\u00e9ri\u00ed<\/li>\r\n \t
  • Extracelul\u00e1rne vezikuly a ich potenci\u00e1lne vyu\u017eitie v klinickej praxi<\/li>\r\n \t
  • Detection of copy number variation from low-coverage whole-genome sequencing data<\/li>\r\n \t
  • Lengths of circulating DNA fragments as a promising predictor of cancer stage<\/li>\r\n \t
  • Vyu\u017eitie hmotnostnej spektrometrie v diagnostike por\u00fach glykozyl\u00e1cie<\/li>\r\n<\/ul>","post_title":"newslab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-7","to_ping":"","pinged":"","post_modified":"2022-02-10 02:23:00","post_modified_gmt":"2022-02-10 01:23:00","post_content_filtered":"","post_parent":0,"guid":"https:\/\/www.newslab.sk\/casopis\/newslab-7\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"69 - 72","upload_clanok":{"ID":2406,"id":2406,"title":"NEWSLAB 2-2021_Pakanova","filename":"NEWSLAB-2-2021_Pakanova-1.pdf","filesize":944868,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2022\/02\/NEWSLAB-2-2021_Pakanova-1.pdf","link":"https:\/\/www.newslab.sk\/en\/the-application-of-mass-spectrometry-in-diagnostics-of-glycosylation-disorders\/newslab-2-2021_pakanova-2-2\/","alt":"","author":"7","description":"","caption":"","name":"newslab-2-2021_pakanova-2-2","status":"inherit","uploaded_to":2408,"date":"2022-02-09 13:27:32","modified":"2022-02-09 13:27:32","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\/2408","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=2408"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/2408\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=2408"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=2408"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=2408"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}