Ukuhlolwa kwe-DNA methylation kuhlanganiswe nama-smartphone ukuze kuhlolwe izimila kusenesikhathi kanye nokuhlolwa kwe-leukemia ngokunemba okungu-90.0%!

Ukutholwa komdlavuza kusenesikhathi ngokusekelwe ku-liquid biopsy kuyindlela entsha yokuthola nokuxilongwa komdlavuza okuphakanyiswe yi-US National Cancer Institute eminyakeni yamuva nje, ngenhloso yokuthola umdlavuza kusenesikhathi noma ngisho nezilonda ezingaphambi komdlavuza. Kusetshenziswe kabanzi njenge-biomarker entsha yokutholakala kwasekuqaleni kwezinhlobo ezahlukene zomdlavuza, okuhlanganisa umdlavuza wamaphaphu, izimila zamathumbu, ama-glioma kanye nezimila zabesifazane.

Ukuvela kwamapulatifomu okuhlonza ama-biomarker e-methylation landscape (Methylscape) kunamandla okuthuthukisa kakhulu ukuhlolwa kwasekuqaleni komdlavuza okukhona, okubeka iziguli esigabeni sokuqala sokwelashwa.

Muva nje, abacwaningi bathuthukise ipulatifomu elula neqondile yokuzwa yokutholwa kwe-methylation landscape ngokusekelwe kuma-nanoparticles egolide ahlotshiswe nge-cysteamine (ama-Cyst/AuNP) ahlanganiswe ne-biosensor esekwe kuma-smartphone evumela ukuhlolwa okusheshayo kwangaphambi kwesikhathi kwezinhlobo eziningi zezimila. Ukuhlolwa kwangaphambi kwesikhathi kwe-leukaemia kungenziwa zingakapheli imizuzu eyi-15 ngemuva kokukhishwa kwe-DNA kusampula yegazi, ngokunemba okungu-90.0%. Isihloko sendatshana sithi Ukutholwa okusheshayo kwe-DNA yomdlavuza egazini lomuntu kusetshenziswa ama-AuNP ambozwe yi-cysteamine kanye ne-smartphone enikwe amandla okufunda ngomshini。

Umfanekiso 1. Ipulatifomu elula nesheshayo yokuhlola umdlavuza ngezingxenye ze-Cyst/AuNPs ingafezwa ngezinyathelo ezimbili ezilula.

Lokhu kuboniswe kuMfanekiso 1. Okokuqala, kwasetshenziswa isisombululo samanzi ukuncibilikisa izingcezu ze-DNA. Ama-Cyst/AuNP abe esengezwa esixazululweni esixubile. I-DNA evamile neyimbi inezakhiwo ezahlukene ze-methylation, okuholela ezingcezu ze-DNA ezinamaphethini ahlukene okuzihlanganisa. I-DNA evamile iyahlangana kancane futhi ekugcineni ihlanganise ama-Cyst/AuNP, okuholela ekushintsheni okubomvu kwama-Cyst/AuNPs, ukuze ushintsho lombala kusuka kokubomvu kuya kokunsomi lubonwe ngamehlo enyama. Ngokuphambene nalokho, iphrofayili ye-methylation ehlukile ye-DNA yomdlavuza iholela ekukhiqizweni kwamaqoqo amakhulu ezingcezu ze-DNA.

Izithombe zamapuleti angu-96-well zathathwa kusetshenziswa ikhamera ye-smartphone. I-DNA yomdlavuza yalinganiswa nge-smartphone ehlonyiswe ngokufunda komshini uma kuqhathaniswa nezindlela ezisekelwe ku-spectroscopy.

Ukuhlolwa komdlavuza kumasampula egazi langempela

Ukuze bandise ukusetshenziswa kwepulatifomu yokuzwa, abacwaningi basebenzise inzwa ehlukanisa ngempumelelo phakathi kwe-DNA evamile neyomdlavuza kumasampula egazi langempela. Amaphethini e-methylation ezindaweni ze-CpG alawula ukuvezwa kwezakhi zofuzo nge-epigenetically. Cishe kuzo zonke izinhlobo zomdlavuza, izinguquko ku-methylation ye-DNA kanye nokubonakaliswa kwezakhi zofuzo ezikhuthaza i-tumorigenesis kuye kwabonwa kushintshana.

Njengesibonelo seminye imidlavuza ehlotshaniswa ne-DNA methylation, abacwaningi basebenzise amasampula egazi avela ezigulini ezine-leukaemia kanye nezilawuli ezinempilo ukuze bahlole ukusebenza kahle kwesimo se-methylation ekuhlukaniseni imidlavuza ye-leukaemia. Le biomarker yesimo se-methylation ayigcini nje ngokudlula izindlela zokuhlola i-leukaemia ezikhona ngokushesha, kodwa futhi ikhombisa ukuthi kungenzeka yini ukusabalala ekutholakaleni kusenesikhathi kwezinhlobo eziningi zomdlavuza kusetshenziswa lolu vivinyo olulula noluqondile.

Kuhlaziywe i-DNA evela kumasampula egazi avela ezigulini ezingama-31 ze-leukaemia kanye nabantu abayi-12 abaphilile. Njengoba kuboniswe ebhokisini ku-Figure 2a, ukumuncwa okuhlobene kwamasampula omdlavuza (ΔA650/525) kwakuphansi kunokwe-DNA evela kumasampula ajwayelekile. Lokhu kwakubangelwa kakhulu ukungezwani kwamanzi okuholela ekuhlanganisweni okukhulu kwe-DNA yomdlavuza, okwavimbela ukuhlanganiswa kwama-Cyst/AuNPs. Ngenxa yalokho, lawa ma-nanoparticles ahlakazeka ngokuphelele ezingqimbeni zangaphandle zama-aggregates omdlavuza, okwaholela ekusakazweni okuhlukile kwama-Cyst/AuNPs afakwe kuma-aggregates e-DNA evamile kanye neyomdlavuza. Ama-ROC curves abe esekhiqizwa ngokushintsha umkhawulo kusukela enanini eliphansi le-ΔA650/525 kuya enanini eliphezulu.

Umfanekiso 2.(a) Amanani okumuncwa okuhlobene kwezixazululo ze-cyst/AuNPs abonisa ukuba khona kwe-DNA evamile (eluhlaza okwesibhakabhaka) kanye nomdlavuza (obomvu) ngaphansi kwezimo ezilungiselelwe kahle

(DA650/525) yamabhokisi; (b) Ukuhlaziywa kwe-ROC nokuhlolwa kokuhlolwa kokuxilonga. (c) I-matrix yokudideka yokuxilonga iziguli ezivamile kanye nezomdlavuza. (d) Ukuzwela, ukucacisa, inani elihle lokubikezela (PPV), inani elibi lokubikezela (NPV) kanye nokunemba kwendlela ethuthukisiwe.

Njengoba kuboniswe kuMfanekiso 2b, indawo engaphansi kwejika le-ROC (AUC = 0.9274) etholwe inzwa ethuthukisiwe ibonise ukuzwela okuphezulu kanye nokucaciswa. Njengoba kungabonakala ebhokisini, iphuzu eliphansi kakhulu elimele iqembu le-DNA elijwayelekile alihlukanisiwe kahle nephuzu eliphakeme kakhulu elimele iqembu le-DNA lomdlavuza; ngakho-ke, ukuhlehla kwe-logistic kwasetshenziswa ukuhlukanisa phakathi kwamaqembu ajwayelekile nalawo anomdlavuza. Njengoba kunikezwe isethi yeziguquguquko ezizimele, ilinganisa amathuba okwenzeka kwesigameko, njengomdlavuza noma iqembu elijwayelekile. I-variable exhomeke kuyo iphakathi kuka-0 no-1. Ngakho-ke umphumela uyindlela engenzeka. Sithole amathuba okuhlonza umdlavuza (P) ngokusekelwe ku-ΔA650/525 kanje.

lapho b=5.3533,w1=-6.965. Ekuhlukanisweni kwesampula, amathuba angaphansi kuka-0.5 abonisa isampula evamile, kanti amathuba angu-0.5 noma ngaphezulu abonisa isampula yomdlavuza. Isibalo 2c sibonisa i-matrix yokudideka ekhiqizwe kusukela ekuqinisekisweni kwe-leave-it-alone cross, okwasetshenziswa ukuqinisekisa ukuzinza kwendlela yokuhlukanisa. Isibalo 2d sifingqa ukuhlolwa kokuhlolwa kokuxilonga kwendlela, kufaka phakathi ukuzwela, ukucacisa, inani elihle lokubikezela (PPV) kanye nenani elibi lokubikezela (NPV).

Ama-biosensor asekelwe kuma-smartphone

Ukuze kwenziwe lula ukuhlolwa kwesampula ngaphandle kokusebenzisa ama-spectrophotometer, abacwaningi basebenzise ubuhlakani bokwenziwa (AI) ukuze bachaze umbala wesisombululo futhi bahlukanise phakathi kwabantu abavamile nabanomdlavuza. Ngenxa yalokhu, umbono wekhompyutha wasetshenziswa ukuhumusha umbala wesisombululo se-Cyst/AuNPs ube yi-DNA evamile (onsomi) noma i-DNA enomdlavuza (obomvu) kusetshenziswa izithombe zamapuleti angu-96-well athathwe ngekhamera yefoni ephathekayo. Ubuhlakani bokwenziwa bunganciphisa izindleko futhi buthuthukise ukufinyeleleka ekuchazeni umbala wezixazululo ze-nanoparticle, futhi ngaphandle kokusebenzisa noma yiziphi izesekeli ze-smartphone zehadiwe ye-optical. Okokugcina, amamodeli amabili okufunda komshini, kufaka phakathi i-Random Forest (RF) kanye ne-Support Vector Machine (SVM) aqeqeshwe ukwakha amamodeli. Womabili amamodeli e-RF kanye ne-SVM ahlukanisa kahle amasampula njengahle nangalungile ngokunemba okungu-90.0%. Lokhu kusikisela ukuthi ukusetshenziswa kobuhlakani bokwenziwa ekuzweleni i-biosensing okusekelwe kumakhalekhukhwini kungenzeka impela.

Umfanekiso 3.(a) Isigaba esiqondiwe sesisombululo esiqoshwe ngesikhathi sokulungiselela isampula yesinyathelo sokuthola isithombe. (b) Isibonelo sesithombe esithathwe ngesikhathi sesinyathelo sokuthola isithombe. (c) Ubukhulu bombala wesisombululo se-cyst/AuNPs emthonjeni ngamunye wepuleti lemithombo engu-96 ekhishwe esithombeni (b).

Besebenzisa ama-Cyst/AuNPs, abacwaningi baphumelele ukuthuthukisa ipulatifomu elula yokuzwa yokuthola indawo ye-methylation kanye nenzwa ekwazi ukuhlukanisa i-DNA evamile ku-DNA yomdlavuza lapho besebenzisa amasampula egazi langempela ukuhlola i-leukaemia. Inzwa ethuthukisiwe ibonise ukuthi i-DNA ekhishwe kumasampula egazi langempela ikwazile ukubona ngokushesha nangokungabizi inani elincane le-DNA yomdlavuza (3nM) ezigulini ezine-leukaemia ngemizuzu eyi-15, futhi ibonise ukunemba okungu-95.3%. Ukuze kube lula kakhulu ukuhlolwa kwesampula ngokuqeda isidingo se-spectrophotometer, ukufunda komshini kusetshenziswe ukuhumusha umbala wesisombululo nokuhlukanisa phakathi kwabantu abavamile nabanomdlavuza kusetshenziswa isithombe sefoni ephathekayo, futhi ukunemba nakho kwakwazi ukufezwa ku-90.0%.

Ireferensi: DOI: 10.1039/d2ra05725e