Kuongororwa kweDNA methylation pamwe chete nemafoni efoni kuti pave nekuongororwa bundu nekukurumidza uye kuongororwa leukaemia ne90.0% chaiyo!

Kuonekwa kwegomarara nekukurumidza zvichibva pakushandiswa kwe liquid biopsy inzira itsva yekuona nekuongorora gomarara yakakurudzirwa neUS National Cancer Institute mumakore achangopfuura, nechinangwa chekuona gomarara nekukurumidza kana kunyange maronda asati asvika kenza. Yave ichishandiswa zvakanyanya sechiratidzo chitsva chekuongorora gomarara rakasiyana-siyana nekukurumidza, kusanganisira gomarara remapapu, gomarara remudumbu, magliomas uye gomarara revakadzi.

Kubuda kwemapuratifomu ekuona methylation landscape (Methylscape) biomarkers kune mukana wekuvandudza zvakanyanya kuongororwa kwegomarara kuripo kare, zvichiita kuti varwere vasvike padanho rekutanga rekurapwa.

Munguva pfupi yapfuura, vaongorori vakagadzira nzira iri nyore uye yakananga yekuona methylation landscape decorated nanoparticles dzegoridhe (Cyst/AuNPs) dzakasanganiswa ne biosensor inoshandiswa pamafoni inoita kuti pave nekukasira kuongororwa kwemhando dzakasiyana-siyana dzemabundu. Kuongorora leukaemia kunogona kuitwa mukati memaminitsi gumi nemashanu mushure mekutorwa kweDNA kubva muropa, nekunyatsojeka kwe90.0%. Musoro wenyaya unoti Kuongororwa nekukurumidza kweDNA yekenza muropa revanhu uchishandisa cysteamine-capped AuNPs uye smartphone inodzidzira muchina.

Mufananidzo 1. Nzvimbo iri nyore uye inokurumidza yekuongorora kenza kuburikidza nezvikamu zveCyst/AuNPs inogona kuitwa nematanho maviri akareruka.

Izvi zvinoratidzwa muMufananidzo 1. Kutanga, mvura yakashandiswa kunyungudutsa zvidimbu zveDNA. Cyst/AuNPs dzakazowedzerwa mumusanganiswa wakasanganiswa. DNA yakajairika neyakaipa ine hunhu hwakasiyana hwemethylation, zvichikonzera zvidimbu zveDNA zvine mapatani akasiyana ekuzvisanganisa. DNA yakajairika inoungana zvisina kurongeka uye pakupedzisira inounganidza Cyst/AuNPs, izvo zvinoguma nekushanduka kweCyst/AuNPs, kuitira kuti shanduko yeruvara kubva kutsvuku kuenda kupepuru ionekwe nemaziso asina kupfeka. Kusiyana neizvi, chimiro chemethylation chakasiyana cheDNA yekenza chinotungamira mukugadzirwa kwezvikamu zvakakura zvezvidimbu zveDNA.

Mifananidzo yemaplates emvura ane matsime makumi mapfumbamwe nematanhatu yakatorwa pachishandiswa kamera yefoni. DNA yekenza yakayerwa nefoni ine michina yekudzidza muchina zvichienzaniswa nenzira dzinoshandiswa pakuongorora spectroscopy.

Kuongororwa kwegomarara muropa chairo

Kuti vawedzere kushanda kwepuratifomu yekunzwa, vaongorori vakashandisa sensor yakabudirira kusiyanisa pakati peDNA yakajairika neyekenza mumasampuli eropa chaiwo. Maitiro emethylation panzvimbo dzeCpG anodzora epigenetically expression yemajini. Mumhando dzese dzekenza, shanduko muDNA methylation uye nekudaro mukutaura kwemajini anokurudzira tumorigenesis zvakaonekwa zvichichinjana.

Semuenzaniso wemamwe magomarara ane chekuita neDNA methylation, vaongorori vakashandisa sampuro dzeropa kubva kuvarwere vane leukaemia pamwe nevanoona nezveutano kuti vaongorore kushanda kwenzvimbo yemethylation mukusiyanisa magomarara eleukaemic. Iyi methylation landscape biomarker haingopfuuri nzira dziripo dzekuongorora leukaemia nekukurumidza, asiwo inoratidza kugona kwekuwedzera kusvika pakuona kenza dzakasiyana-siyana uchishandisa iyi assay iri nyore uye iri nyore.

DNA kubva muropa revarwere vane leukaemia makumi matatu nerimwe nevanhu gumi nevaviri vane hutano yakaongororwa. Sezvakaratidzwa mubhokisi riri muMufananidzo 2a, kunyudzwa kwemasampuli ekenza (ΔA650/525) kwaive kwakaderera pane kweDNA kubva musampuli dzakajairika. Izvi zvainyanya kukonzerwa nekusagadzikana kwemvura kwakakonzera kuunganidzwa kwakasimba kweDNA yekenza, izvo zvakadzivirira kuunganidzwa kweCyst/AuNPs. Nekuda kweizvozvo, idzi nanoparticles dzakapararira zvachose muzvikamu zvekunze zvegomarara, izvo zvakakonzera kupararira kwakasiyana kweCyst/AuNPs dzakanamirwa pagomarara reDNA regomarara neregomarara. Makove eROC akazogadzirwa nekusiyanisa chiyero kubva pamutengo wakaderera weΔA650/525 kusvika pamutengo wakanyanya.

Mufananidzo 2.(a) Kuwanda kwemakemikari anowanikwa mu cyst/AuNPs kunoratidza kuvapo kweDNA yakajairika (yebhuruu) uye kenza (tsvuku) mumamiriro ezvinhu akagadziriswa.

(DA650/525) yemabhokisi ekuongorora; (b) Kuongorora kweROC uye kuongorora bvunzo dzekuongorora. (c) Matrix yekuvhiringidzika kwekuongorora varwere vakagwinya nekenza. (d) Kunzwa, kujekesa, kukosha kwekufungidzira kwakanaka (PPV), kukosha kwekufungidzira kusina kunaka (NPV) uye kururama kwenzira yakagadzirwa.

Sezvakaratidzwa paMufananidzo 2b, nzvimbo iri pasi peROC curve (AUC = 0.9274) yakawanikwa kune sensor yakagadzirwa yakaratidza kunzwisisika kwakanyanya uye hunyanzvi. Sezvazvinogona kuonekwa kubva pabhokisi rebhokisi, nzvimbo yakaderera inomiririra boka reDNA rakajairika haina kuparadzaniswa zvakanaka nenzvimbo yepamusoro inomiririra boka reDNA rekenza; saka, logistic regression yakashandiswa kusiyanisa pakati pemapoka akajairika neekenza. Zvichipiwa seti yezvimwe zvinhu zvakazvimiririra, inofungidzira mukana wekuti chiitiko chiitike, senge kenza kana boka rakajairika. Iyo dependent variable iri pakati pe0 ne1. Saka mhedzisiro yacho mukana. Takaona mukana wekuziva kenza (P) zvichibva paΔA650/525 seizvi.

apo b=5.3533,w1=-6.965. Pakupatsanura sampuli, mukana uri pasi pe0.5 unoratidza muenzaniso wakajairika, nepo mukana we0.5 kana kupfuura uchiratidza muenzaniso wekenza. Mufananidzo 2c unoratidza matrix yekuvhiringidzika yakagadzirwa kubva mu "leave-it-alone cross-validation", iyo yakashandiswa kusimbisa kugadzikana kwenzira yekupatsanura. Mufananidzo 2d unopfupikisa kuongororwa kwebvunzo yekuongorora nzira, kusanganisira sensitivity, specificity, positive predictive value (PPV) uye negative predictive value (NPV).

Masensa e biosensors akavakirwa pamafoni

Kuti vawedzere kurerutsa kuyedzwa kwemuenzaniso pasina kushandisa ma spectrophotometer, vaongorori vakashandisa njere dzekugadzira (AI) kududzira ruvara rwemhinduro uye kusiyanisa pakati pevanhu vakajairwa nekenza. Zvichitevera izvi, kuona kwekombiyuta kwakashandiswa kushandura ruvara rwemhinduro yeCyst/AuNPs kuita DNA yakajairika (yepepuru) kana DNA yekenza (tsvuku) vachishandisa mifananidzo yemaplates e96-well akatorwa kuburikidza nekamera yefoni. Njere dzekugadzira dzinogona kuderedza mitengo uye kuvandudza mukana wekududzira ruvara rwemhinduro dze nanoparticle, uye pasina kushandisa chero zvishongedzo zvefoni zve optical hardware. Pakupedzisira, mamodheru maviri ekudzidza emuchina, anosanganisira Random Forest (RF) neSupport Vector Machine (SVM) akadzidziswa kugadzira mamodheru. mamodheru eRF neSVM ese akaronga mamodheru acho seakanaka neasina kunaka ne90.0%. Izvi zvinoratidza kuti kushandiswa kwenjere dzekugadzira mu biosensing inoshandiswa pafoni kunogoneka.

Mufananidzo 3.(a) Kirasi yakatarwa yemhinduro yakanyorwa panguva yekugadzirira sampuro yedanho rekutora mufananidzo. (b) Muenzaniso wemufananidzo wakatorwa panguva yedanho rekutora mufananidzo. (c) Kusimba kweruvara rwe cyst/AuNPs solution mutsime rega rega replate ye 96-well yakatorwa kubva pamufananidzo (b).

Vachishandisa Cyst/AuNPs, vaongorori vakabudirira kugadzira puratifomu iri nyore yekuona methylation landscape uye sensor inokwanisa kusiyanisa DNA yakajairika kubva kuDNA yekenza pavanoshandisa sampuli dzeropa chaidzo kuongorora leukaemia. Sensor yakagadzirwa yakaratidza kuti DNA yakatorwa kubva musampuli dzeropa chaidzo yakakwanisa kuona nekukurumidza uye zvine mutengo mudiki diki dzeDNA yekenza (3nM) muvarwere vane leukaemia mumaminitsi gumi nemashanu, uye yakaratidza kururama kwe95.3%. Kuti zvinyanye kurerutsa kuyedzwa kwemuenzaniso nekubvisa kudiwa kwespectrophotometer, kudzidza kwemuchina kwakashandiswa kududzira ruvara rwemhinduro uye kusiyanisa pakati pevanhu vakajairwa nekenza vachishandisa mufananidzo wefoni, uye kururama kwakakwanisawo kuwanikwa pa90.0%.

Referensi: DOI: 10.1039/d2ra05725e