Uvavanyo lwe-DNA methylation kunye neefowuni eziphathwayo zokuhlola iithumba kwangethuba kunye nokuhlolwa kwe-leukaemia ngokuchanekileyo kwe-90.0%!

Ukufunyanwa komhlaza kwangethuba ngokusekelwe kwi-liquid biopsy yindlela entsha yokufumanisa nokuchonga umhlaza ecetywayo yi-US National Cancer Institute kwiminyaka yakutshanje, ngenjongo yokufumanisa umhlaza kwangethuba okanye izilonda ezingaphambi komhlaza. Isetyenziswe kakhulu njenge-biomarker entsha yokufumanisa kwangethuba iintlobo ngeentlobo ze-malignancies, kubandakanya umhlaza wemiphunga, ii-tumor zesisu, ii-gliomas kunye nee-tumor ze-gynaecological.

Ukuvela kwamaqonga okuchonga ii-biomarkers ze-methylation landscape (Methylscape) kunamandla okuphucula kakhulu ukuvavanywa komhlaza kwangethuba okukhoyo, okubeka izigulana kwinqanaba lokuqala lokunyangwa.

Kutshanje, abaphandi baye baphuhlisa iqonga elilula nelithe ngqo lokufumanisa i-methylation landscape ngokusekelwe kwi-nanoparticles zegolide ezihonjiswe yi-cysteamine (iiCyst/AuNPs) ezidityaniswe ne-biosensor esekwe kwi-smartphone evumela ukuhlolwa ngokukhawuleza kwangethuba kweentlobo ezahlukeneyo zeethumba. Ukuhlolwa kwangethuba kwe-leukaemia kungenziwa kwimizuzu eli-15 emva kokukhutshwa kwe-DNA kwisampulu yegazi, ngokuchanekileyo kwe-90.0%. Isihloko senqaku ngu-Ukuchongwa ngokukhawuleza kwe-DNA yomhlaza egazini lomntu kusetyenziswa ii-AuNPs ezine-cysteamine kunye ne-smartphone enikwe amandla okufunda ngomatshini.

Umfanekiso 1. Iqonga elilula nelikhawulezayo lokuvavanya umhlaza ngokusebenzisa izixhobo zeCyst/AuNPs linokwenziwa ngamanyathelo amabini alula.

Oku kuboniswe kuMfanekiso 1. Okokuqala, isisombululo samanzi sasetyenziswa ukunyibilikisa iziqwenga zeDNA. Emva koko kongezwa iiCyst/AuNPs kwisisombululo esixutyiweyo. I-DNA eqhelekileyo neyimbi ineempawu ezahlukeneyo zemethylation, nto leyo ebangela iziqwenga zeDNA ezineepateni ezahlukeneyo zokuzihlanganisa. I-DNA eqhelekileyo iyahlangana kwaye ekugqibeleni ihlanganise iiCyst/AuNPs, nto leyo ebangela ukuba iiCyst/AuNPs zitshintshe umbala, ukuze utshintsho lombala ukusuka kubomvu ukuya kumfusa lubonwe ngamehlo enyama. Ngokwahlukileyo koko, iprofayili yemethylation eyahlukileyo yeDNA yomhlaza ikhokelela ekuvelisweni kwamaqela amakhulu eziqwenga zeDNA.

Imifanekiso yeepleyiti ezingama-96-well ithathwe kusetyenziswa ikhamera yefowuni. I-DNA yomhlaza ilinganiswe ngefowuni exhotyiswe ngokufunda komatshini xa kuthelekiswa neendlela ezisekelwe kwi-spectroscopy.

Ukuhlolwa komhlaza kwiisampuli zegazi lokwenyani

Ukwandisa ukusetyenziswa kweqonga lokuva, abaphandi basebenzise isenzi esahlula ngempumelelo phakathi kwe-DNA eqhelekileyo neyomhlaza kwiisampuli zegazi lokwenyani. Iipateni ze-methylation kwiindawo ze-CpG zilawula ukubonakaliswa kwezakhi zofuzo. Phantse kuzo zonke iintlobo zomhlaza, utshintsho kwi-methylation ye-DNA kwaye ngaloo ndlela ekubonakalisweni kwezakhi zofuzo ezikhuthaza i-tumorigenesis ziye zabonwa zitshintshatshintsha.

Njengomzekelo weminye i-cancer enxulumene ne-DNA methylation, abaphandi basebenzise iisampuli zegazi ezivela kwizigulane ezine-leukaemia kunye nabalawuli abasempilweni ukuphanda ukusebenza kakuhle kwe-methylation landscape ekwahlukaniseni umhlaza we-leukaemic. Le biomarker ye-methylation landscape ayipheleli nje ekuphumeleleni iindlela zokuhlola i-leukaemia ezikhawulezayo ezikhoyo, kodwa ikwabonisa ukuba kunokwenzeka ukufumanisa uluhlu olubanzi lomhlaza kwangethuba kusetyenziswa olu vavanyo lulula nolucacileyo.

I-DNA evela kwiisampuli zegazi ezivela kwizigulana ezingama-31 ze-leukaemia kunye nabantu abali-12 abaphilileyo ihlalutywe. njengoko kubonisiwe kwiploti yebhokisi kuMfanekiso 2a, ukufunxwa kweesampuli zomhlaza (ΔA650/525) bekuphantsi kunokwe-DNA evela kwiisampuli eziqhelekileyo. Oku kwakubangelwa ikakhulu kukungazinzi kwamanzi okukhokelela ekuhlanganisweni okukhulu kwe-DNA yomhlaza, okwathintela ukuhlanganiswa kwee-Cyst/AuNPs. Ngenxa yoko, ezi nanoparticles zasasazeka ngokupheleleyo kwiingqimba zangaphandle zee-aggregates zomhlaza, nto leyo eyabangela ukusasazeka okwahlukileyo kwe-Cyst/AuNPs ezifakwe kwii-aggregates ze-DNA eziqhelekileyo kunye nezomhlaza. Ii-ROC curves emva koko zenziwa ngokutshintsha umda ukusuka kwixabiso elincinci le-ΔA650/525 ukuya kwixabiso eliphezulu.

Umfanekiso 2.(a) Amaxabiso okufunxwa okunxulumeneyo kwezisombululo ze-cyst/AuNPs abonisa ubukho be-DNA eqhelekileyo (eluhlaza okwesibhakabhaka) kunye nomhlaza (obomvu) phantsi kweemeko ezilungisiweyo.

(DA650/525) yeeploti zebhokisi; (b) Uhlalutyo lwe-ROC kunye novavanyo lweemvavanyo zokuxilonga. (c) I-Confusion matrix yokuxilonga izigulane eziqhelekileyo kunye nezomhlaza. (d) Uvakalelo, ukucaca, ixabiso eliqikelelweyo elihle (PPV), ixabiso elibi lokuqikeleleka (NPV) kunye nokuchaneka kwendlela ephuhlisiweyo.

Njengoko kubonisiwe kuMfanekiso 2b, indawo engaphantsi kwe-ROC curve (AUC = 0.9274) efunyenwe kwi-sensor ephuhlisiweyo ibonise uvakalelo oluphezulu kunye nokucaciswa. Njengoko kunokubonwa kwiploti yebhokisi, indawo esezantsi emele iqela le-DNA eliqhelekileyo alahlulwanga kakuhle kwindawo ephezulu emele iqela le-DNA lomhlaza; ke ngoko, i-logistic regression yasetyenziswa ukwahlula phakathi kwamaqela aqhelekileyo kunye namaqela omhlaza. Ngenxa yesethi yeenguqu ezizimeleyo, iqikelela amathuba okuba kwenzeke isiganeko, njengomhlaza okanye iqela eliqhelekileyo. Inguqu exhomekeke kuyo iphakathi kwe-0 kunye ne-1. Isiphumo ke ngoko sinokwenzeka. Simisele amathuba okuchongwa komhlaza (P) ngokusekelwe kwi-ΔA650/525 ngolu hlobo lulandelayo.

apho b=5.3533,w1=-6.965. Kwindlela yokwahlulahlula isampuli, amathuba angaphantsi kwe-0.5 abonisa isampuli eqhelekileyo, ngelixa amathuba angama-0.5 okanye ngaphezulu abonisa isampuli yomhlaza. Umfanekiso 2c ubonisa i-confusion matrix eyenziwe kwi-leave-it-alone cross-validation, eyasetyenziswa ukuqinisekisa uzinzo lwendlela yokwahlulahlula. Umfanekiso 2d ushwankathela uvavanyo lovavanyo lokuxilonga lwendlela, kubandakanya uvakalelo, ukucaca, ixabiso eliqikelelweyo elilungileyo (PPV) kunye nexabiso elibi lokuqikelela (NPV).

Ii-biosensors ezisekelwe kwi-smartphone

Ukuze kube lula ngakumbi ukuvavanya iisampulu ngaphandle kokusebenzisa ii-spectrophotometers, abaphandi basebenzise i-artificial intelligence (AI) ukutolika umbala wesisombululo nokwahlula phakathi kwabantu abaqhelekileyo nabanesifo somhlaza. Ngenxa yoku, umbono wekhompyutha usetyenzisiwe ukuguqulela umbala wesisombululo seCyst/AuNPs kwi-DNA eqhelekileyo (emfusa) okanye i-DNA enomhlaza (ebomvu) kusetyenziswa imifanekiso yeepleyiti ezingama-96 ezithathwe ngekhamera yefowuni ephathwayo. I-artificial intelligence inokunciphisa iindleko kwaye iphucule ukufikeleleka ekutolikeni umbala wezisombululo ze-nanoparticle, kwaye ngaphandle kokusebenzisa naziphi na izixhobo ze-smartphone ze-optical hardware. Okokugqibela, iimodeli ezimbini zokufunda koomatshini, kuquka iRandom Forest (RF) kunye neSupport Vector Machine (SVM) zaqeqeshwa ukwakha iimodeli. zombini iimodeli zeRF kunye ne-SVM zahlula ngokuchanekileyo iisampulu njengezakhayo nezingalunganga ngokuchanekileyo kwe-90.0%. Oku kuthetha ukuba ukusetyenziswa kwe-artificial intelligence kwi-biosensing esekelwe kwifowuni ephathwayo kunokwenzeka.

Umfanekiso 3.(a) Udidi olujoliswe kulo lwesisombululo olurekhodwe ngexesha lokulungiselela isampuli yenyathelo lokufumana umfanekiso. (b) Umzekelo womfanekiso othathwe ngexesha lenyathelo lokufumana umfanekiso. (c) Ubungakanani bombala wesisombululo se-cyst/AuNPs kwiqula ngalinye lepleyiti ye-96-well ekhutshwe kumfanekiso (b).

Besebenzisa iiCyst/AuNPs, abaphandi baphumelele ekuphuhliseni iqonga elilula lokubona i-methylation landscape kunye nesensor ekwaziyo ukwahlula i-DNA eqhelekileyo kwi-DNA yomhlaza xa besebenzisa iisampuli zegazi zokwenyani ukuhlola i-leukaemia. Le sensor iphuhlisiweyo ibonise ukuba i-DNA ekhutshwe kwiisampuli zegazi zokwenyani ikwazile ukubona ngokukhawuleza nangokufanelekileyo inani elincinci le-DNA yomhlaza (3nM) kwizigulane ezine-leukaemia kwimizuzu eli-15, kwaye ibonise ukuchaneka kwe-95.3%. Ukuze kube lula ngakumbi ukuvavanywa kwesampulu ngokuphelisa imfuneko ye-spectrophotometer, ukufunda komatshini kusetyenzisiwe ukutolika umbala wesisombululo kunye nokwahlula phakathi kwabantu abaqhelekileyo nabanomhlaza kusetyenziswa ifoto yefowuni ephathwayo, kwaye ukuchaneka nako kwakwazi ukuphunyezwa kwi-90.0%.

Isalathiso: DOI: 10.1039/d2ra05725e