labarai

A cikin shekaru goma da suka gabata, an yi amfani da fasahar sarrafa kwayoyin halitta a cikin binciken kansa da kuma aikin asibiti, ya zama muhimmin kayan aiki don bayyana halayen kwayoyin cutar kansa. Ci gaba a cikin ganewar ƙwayoyin ƙwayoyin cuta da maganin da aka yi niyya sun haɓaka haɓakar ra'ayoyin jiyya na ƙwayar ƙwayar cuta kuma ya kawo babban canje-canje ga duka fannin gano cutar kansa da jiyya. Ana iya amfani da gwajin kwayoyin halitta don gargaɗin haɗarin ciwon daji, jagorar yanke shawara na jiyya da kimanta tsinkaye, kuma kayan aiki ne mai mahimmanci don haɓaka sakamakon asibiti na haƙuri. Anan, mun taƙaita labaran kwanan nan da aka buga a CA Cancer J Clin, JCO, Ann Oncol da sauran mujallu don nazarin aikace-aikacen gwajin kwayoyin halitta a cikin ganewar asali da magani.

Sauye-sauyen somatic da maye gurbi. Gabaɗaya, ciwon daji yana haifar da maye gurbi na DNA wanda za'a iya gada daga iyaye (maye gurbi) ko kuma aka samu tare da shekaru (maye gurbi na somatic). Canje-canjen layin ƙwayoyin cuta suna nan tun daga haihuwa, kuma mutator yawanci yana ɗaukar maye gurbi a cikin DNA na kowane tantanin halitta a jiki kuma ana iya kaiwa ga zuriya. Maye gurbi na somatic ana samun su a cikin sel marasa gamuwa kuma yawanci ba a ba da su ga zuriya ba. Dukansu germline da maye gurbi na somatic na iya lalata ayyukan yau da kullun na sel kuma su haifar da mummunan canji na sel. Maye gurbi na somatic shine babban direba na rashin lafiya kuma mafi yawan tsinkayar kwayoyin halitta a cikin oncology; duk da haka, kusan kashi 10 zuwa 20 na masu ciwon daji suna ɗauke da maye gurbin ƙwayoyin cuta wanda ke ƙara haɗarin cutar kansa sosai, kuma wasu daga cikin waɗannan maye gurbi suna da magani.
maye gurbin direba da fasinja. Ba duk bambance-bambancen DNA ke shafar aikin tantanin halitta ba; a matsakaita, yana ɗaukar abubuwa biyar zuwa goma abubuwan da suka faru na genomic, waɗanda aka sani da “maye gurɓata direba,” don haifar da lalacewa ta al'ada. Sauye-sauyen direbobi yakan faru a cikin kwayoyin halittar da ke da alaƙa da ayyukan rayuwar tantanin halitta, kamar kwayoyin halittar da ke cikin tsarin haɓakar ƙwayoyin cuta, gyaran DNA, sarrafa zagayowar tantanin halitta da sauran hanyoyin rayuwa, kuma suna da yuwuwar a yi amfani da su azaman maƙasudin warkewa. Koyaya, adadin maye gurbi a kowace cutar kansa yana da girma sosai, kama daga ƴan dubbai a wasu cututtukan nono zuwa sama da 100,000 a cikin wasu cututtukan daji masu saurin canza launin launi da na endometrial. Yawancin maye gurbi ba su da ma'anar ilimin halitta ko iyakance, ko da maye gurbi ya faru a yankin coding, irin waɗannan abubuwan maye gurbi marasa mahimmanci ana kiran su "maye gurbi". Idan bambance-bambancen kwayar halitta a cikin wani nau'in ƙari na musamman ya annabta martaninsa ga ko juriya ga jiyya, ana ɗaukar bambance-bambancen a asibiti.
Oncogenes da ƙwayoyin cuta masu hana ƙari. Kwayoyin halittar da ake yawan rikidewa a cikin ciwon daji za a iya kasu kusan kashi biyu, kwayoyin oncogenes da kwayoyin cutar ciwon tumo. A cikin sel na al'ada, sunadaran da ke tattare da oncogenes galibi suna taka rawa na haɓaka haɓakar tantanin halitta da hana apoptosis na tantanin halitta, yayin da furotin da ke tattare da kwayoyin halittar oncosuppressor shine ke da alhakin sarrafa rarrabawar tantanin halitta mara kyau don kula da aikin salula na yau da kullun. A cikin mummunan tsarin canji, maye gurbi na genomic yana haifar da haɓaka ayyukan oncogene da raguwa ko asarar ayyukan kwayoyin oncosuppressor.
Ƙananan bambance-bambance da bambancin tsari. Waɗannan su ne manyan nau'ikan maye gurbi guda biyu a cikin kwayoyin halitta. Ƙananan bambance-bambancen suna canza DNA ta hanyar canzawa, sharewa, ko ƙara ƙaramin adadin tushe, ciki har da saka tushe, gogewa, frameshift, fara asarar codon, dakatar da asarar codon, da dai sauransu. Bambancin tsari shine babban sake tsara kwayoyin halitta, wanda ya ƙunshi sassan kwayoyin halitta masu girma daga 'yan sansanonin dubu kaɗan zuwa mafi yawan adadin kwayoyin halitta, canje-canje a cikin chromosome, ciki har da chromosome na kwafin halitta, ko fassara. Waɗannan maye gurbi na iya haifar da raguwa ko haɓaka aikin furotin. Bugu da ƙari ga canje-canje a matakin jinsin mutum ɗaya, sa hannu na genomic kuma wani ɓangare ne na rahotannin jeri na asibiti. Ana iya ganin sa hannun genomic a matsayin hadaddun sifofi na ƙanana da/ko bambance-bambancen tsari, gami da nauyin maye gurbi (TMB), rashin zaman lafiyar microsatellite (MSI), da lahani na haɗakar juna.

Sauye-sauye na clonal da maye gurbin subclonal. Maye gurbi na clonal suna nan a cikin dukkan ƙwayoyin ƙwayar cuta, suna nan a lokacin ganewar asali, kuma suna kasancewa bayan ci gaban jiyya. Don haka, maye gurbi na clonal yana da yuwuwar a yi amfani da shi azaman maƙasudin warkewar ƙari. Sauye-sauye na subclonal suna samuwa ne kawai a cikin ƙananan ƙwayoyin ciwon daji kuma ana iya gano su a farkon ganewar asali, amma suna ɓacewa tare da maimaitawa na gaba ko bayyana kawai bayan magani. Halin ciwon daji yana nufin kasancewar maye gurbi na subclonal da yawa a cikin kansa guda ɗaya. Musamman ma, galibin sauye-sauyen sauye-sauye na asibiti a cikin duk nau'in ciwon daji na kowa sune maye gurbi na clonal kuma suna dawwama a duk tsawon ci gaban kansa. Juriya, wanda sau da yawa ke shiga tsakani ta subclones, ƙila ba za a iya gano shi ba a lokacin ganewar asali amma yana bayyana lokacin da ya sake komawa bayan jiyya.

Ana amfani da dabarar gargajiya ta FISH ko cell karyotype don gano canje-canje a matakin chromosomal. Ana iya amfani da KIFI don gano haɗaɗɗun kwayoyin halitta, gogewa, da haɓakawa, kuma ana ɗaukarsa a matsayin "ma'aunin zinare" don gano irin waɗannan bambance-bambancen, tare da daidaito mai girma da azanci amma iyakancewar kayan aiki. A wasu cututtukan cututtukan jini, musamman cutar sankarar bargo, har yanzu ana amfani da karyotyping don jagorantar ganewar asali da tsinkaye, amma ana maye gurbin wannan dabarar ta hanyar gwajin kwayoyin da aka yi niyya kamar FISH, WGS, da NGS.
PCR na iya gano canje-canje a cikin kwayoyin halitta guda ɗaya, duka PCR na ainihi da PCR na dijital. Wadannan fasahohin suna da hankali sosai, sun dace musamman don ganowa da lura da ƙananan raunuka, kuma suna iya samun sakamako a cikin ɗan gajeren lokaci, rashin amfani shine cewa iyakar ganowa yana da iyaka (yawanci kawai gano maye gurbi a cikin ɗaya ko ƴan kwayoyin halitta), kuma ikon yin gwaje-gwaje masu yawa yana iyakance.
Immunohistochemistry (IHC) kayan aikin sa ido ne na tushen furotin da aka saba amfani da shi don gano maganganun alamomin halittu kamar ERBB2 (HER2) da masu karɓar isrogen. Hakanan ana iya amfani da IHC don gano takamaiman sunadaran da suka rikiɗe (kamar BRAF V600E) da ƙayyadaddun ƙwayoyin halitta (kamar ALK fusions). Amfanin IHC shine cewa ana iya haɗa shi cikin sauƙi a cikin tsarin nazarin nama na yau da kullun, don haka ana iya haɗa shi tare da wasu gwaje-gwaje. Bugu da ƙari, IHC na iya ba da bayani game da ƙayyadaddun furotin na subcellular. Rashin lahani shine iyakancewar haɓakawa da manyan buƙatun ƙungiyoyi.
Sequencing na ƙarni na biyu (NGS) NGS yana amfani da manyan hanyoyin aiwatar da daidaitattun dabaru don gano bambance-bambance a matakin DNA da/ko RNA. Ana iya amfani da wannan fasaha don jera duka kwayoyin halitta (WGS) da yankuna masu sha'awa. WGS yana ba da mafi cikakkun bayanai na maye gurbi, amma akwai cikas da yawa ga aikace-aikacen sa na asibiti, gami da buƙatar sabbin samfuran ƙwayar ƙwayar cuta (WGS bai dace da nazarin samfuran da ba a iya motsi ba) da tsada.
Jeri na NGS da aka yi niyya ya haɗa da jeri na exon gabaɗaya da kuma rukunin kwayoyin da aka yi niyya. Waɗannan gwaje-gwajen sun wadatar da yankuna masu sha'awa ta hanyar binciken DNA ko haɓakawa na PCR, don haka iyakance adadin jerin abubuwan da ake buƙata (dukkan exome ya ƙunshi kashi 1 zuwa 2 cikin ɗari na genome, har ma da manyan bangarori masu ɗauke da kwayoyin halitta 500 ne kawai kashi 0.1 na kwayoyin halitta). Ko da yake gabaɗayan jerin abubuwan exon yana aiki da kyau a cikin ƙayyadaddun kyallen takarda, farashin sa ya kasance mai girma. Haɗin kwayoyin halittar manufa suna da ɗan ƙarancin tattalin arziki kuma suna ba da damar sassauƙa a zaɓin ƙwayoyin halittar da za a gwada. Bugu da kari, DNA kyauta (cfDNA) ke yawo a matsayin sabon zaɓi don nazarin kwayoyin halitta na masu cutar kansa, wanda aka sani da biopsies na ruwa. Dukkan kwayoyin cutar kansa da kwayoyin halitta na al'ada suna iya sakin DNA cikin jini, kuma DNA da aka zubar daga kwayoyin cutar kansa ana kiranta DNA tumor DNA (ctDNA), wanda za'a iya yin nazari don gano yiwuwar maye gurbi a cikin kwayoyin tumo.
Zaɓin gwajin ya dogara da takamaiman matsalar asibiti da za a magance. Yawancin alamomin halittu masu alaƙa da hanyoyin da aka yarda da su ana iya gano su ta hanyar fasahar KIFI, IHC, da kuma PCR. Wadannan hanyoyin suna da ma'ana don gano ƙananan ƙwayoyin halitta, amma ba su inganta ingantaccen ganewa tare da haɓaka kayan aiki ba, kuma idan an gano yawancin kwayoyin halitta, ƙila ba za a sami isasshen nama don ganowa ba. A wasu takamaiman cututtukan daji, irin su kansar huhu, inda samfuran nama ke da wahala a samu kuma akwai alamun halittu masu yawa don gwadawa, ta amfani da NGS shine mafi kyawun zaɓi. A ƙarshe, zaɓin ƙididdiga ya dogara ne akan adadin masu gano kwayoyin halitta da za a gwada don kowane majiyyaci da kuma adadin marasa lafiya da za a gwada don maganin kwayoyin halitta. A wasu lokuta, yin amfani da IHC/FISH ya wadatar, musamman ma lokacin da aka gano manufa, kamar gano masu karɓar isrogen, masu karɓar progesterone, da ERBB2 a cikin masu ciwon nono. Idan ana buƙatar ƙarin cikakken bincike na maye gurbi da kuma neman yuwuwar maƙasudin warkewa, NGS ta fi tsari da tsada. Bugu da ƙari, ana iya yin la'akari da NGS a cikin lokuta inda sakamakon IHC/FISH ya kasance mai ma'ana ko rashin daidaituwa.

Sharuɗɗa daban-daban suna ba da jagora kan abin da ya kamata marasa lafiya su cancanci gwajin kwayoyin halitta. A cikin 2020, ESMO Precision Medicine Working Group ya ba da shawarwarin gwaji na farko na NGS ga marasa lafiya da ke fama da ciwon daji, suna ba da shawarar gwajin NGS na yau da kullun don ci gaban ciwon huhu mara ƙarancin ƙwayar cuta, ciwon gurgu, ciwon daji, ciwon daji na bile duct, da samfuran ciwon daji na kwai, kuma a cikin 2024, ESMO an sabunta shi da ƙari na ciwon daji. Irin su ciwace-ciwacen hanji na hanji, sarcomas, ciwon daji na thyroid da ciwon daji da ba a san asalinsu ba.
A cikin 2022, Ra'ayin Clinical na ASCO game da gwajin kwayoyin halitta a cikin marasa lafiya da ke fama da ciwon daji ko ciwon daji ya bayyana cewa idan an yarda da maganin da ke da alaƙa da biomarker a cikin marasa lafiya da ke fama da ciwon daji ko ciwace-ciwace, ana ba da shawarar gwajin ƙwayoyin cuta ga waɗannan marasa lafiya. Misali, yakamata a yi gwajin genomic a cikin marasa lafiya tare da melanoma na metastatic don nunawa ga maye gurbi na BRAF V600E, kamar yadda masu hana RAF da MEK suka amince da wannan nuni. Bugu da kari, ya kamata kuma a yi gwajin kwayoyin halitta idan akwai alamar juriya ga maganin da za a yi wa majiyyaci. Egfrmab, alal misali, bashi da tasiri a cikin mutant colorectal cancer KRAS. Lokacin yin la'akari da dacewa da majiyyaci don jerin kwayoyin halitta, ya kamata a haɗa yanayin jiki na majiyyaci, cututtuka, da kuma matakin ciwon daji, saboda jerin matakan da ake bukata don tsarin kwayoyin halitta, ciki har da yarda da haƙuri, aikin dakin gwaje-gwaje, da kuma nazarin sakamakon sakamakon, yana buƙatar mai haƙuri ya sami isasshen ƙarfin jiki da kuma tsawon rai.
Baya ga maye gurbi na somatic, ya kamata a gwada wasu cututtukan daji don ƙwayoyin ƙwayoyin cuta. Gwaji don maye gurbin layin ƙwayoyin cuta na iya yin tasiri ga yanke shawarar jiyya don ciwon daji kamar BRCA1 da maye gurbin BRCA2 a cikin nono, ovarian, prostate, da ciwon daji na pancreatic. Maye gurbi na iya yin tasiri ga gwajin cutar kansa na gaba da rigakafi a cikin marasa lafiya. Marasa lafiya waɗanda ke da yuwuwar dacewa don gwaji don maye gurbin ƙwayoyin cuta suna buƙatar saduwa da wasu sharuɗɗa, waɗanda suka haɗa da abubuwa kamar tarihin iyali na ciwon daji, shekaru a ganewar asali, da nau'in ciwon daji. Duk da haka, yawancin marasa lafiya (har zuwa 50%) suna ɗauke da maye gurbi a cikin layin ƙwayoyin cuta ba su cika ka'idodin gargajiya don gwada maye gurbin layin ƙwayoyin cuta ba bisa tarihin iyali. Don haka, don haɓaka gano masu ɗaukar maye gurbi, Ƙungiyar Ciwon Kankara ta Ƙasa (NCCN) ta ba da shawarar cewa duk ko mafi yawan marasa lafiya masu nono, ovarian, endometrial, pancreatic, colorectal, ko prostate cancer a gwada su don maye gurbin layin kwayoyin cuta.
Game da lokacin gwajin kwayoyin halitta, saboda yawancin sauye-sauyen sauye-sauye na asibiti sun kasance clonal kuma suna da kwanciyar hankali a kan ci gaban ciwon daji, yana da kyau a yi gwajin kwayoyin halitta akan marasa lafiya a lokacin ganewar asali na ciwon daji. Don gwaje-gwajen kwayoyin halitta na gaba, musamman bayan maganin da aka yi niyya na kwayoyin halitta, gwajin ctDNA ya fi fa'ida fiye da ƙwayar ƙwayar cuta ta DNA, saboda DNA na jini na iya ƙunsar DNA daga duk raunukan ƙari, wanda ya fi dacewa don samun bayanai game da nau'in ƙwayar cuta.
Binciken ctDNA bayan jiyya na iya iya yin hasashen martanin ƙari ga jiyya da gano ci gaban cuta a baya fiye da daidaitattun hanyoyin hoto. Duk da haka, ba a kafa ka'idoji don amfani da waɗannan bayanan don jagorantar shawarwarin jiyya ba, kuma ba a ba da shawarar yin nazarin ctDNA ba sai dai a gwaji na asibiti. Hakanan za'a iya amfani da ctDNA don tantance ƙananan raunukan da suka rage bayan tiyatar ƙari. Gwajin ctDNA bayan tiyata shine ma'auni mai ƙarfi na ci gaban cututtuka na gaba kuma yana iya taimakawa wajen sanin ko majiyyaci zai amfana daga chemotherapy adjuvant, amma har yanzu ba a ba da shawarar yin amfani da ctDNA a waje da gwaje-gwaje na asibiti don jagorantar yanke shawara na chemotherapy adjuvant.

Gudanar da bayanai Mataki na farko a cikin jerin kwayoyin halitta shine cire DNA daga samfuran marasa lafiya, shirya dakunan karatu, da samar da cikakkun bayanan jeri. Danyen bayanan yana buƙatar ƙarin aiki, gami da tace bayanai marasa inganci, kwatanta shi tare da genome na tunani, gano nau'ikan maye gurbi ta hanyar algorithms daban-daban, ƙayyadaddun tasirin waɗannan maye gurbi akan fassarar furotin, da tace maye gurbin layin ƙwayoyin cuta.
An ƙirƙira bayanin halittar direba don bambanta maye gurbin direba da fasinja. Sauye-sauyen direbobi suna haifar da asara ko haɓaka ayyukan ƙwayoyin cuta masu hana ƙari. Ƙananan bambance-bambancen da ke haifar da rashin kunna ƙwayoyin ƙwayoyin cuta na ƙwayar cuta sun haɗa da maye gurbi na banza, maye gurbi na firamshift, da maye gurbi na maɓalli, da ƙarancin fara shafewar codon, dakatar da shafewar codon, da kewayon shigar/share maye gurbi. Bugu da ƙari, ƙetare maye gurbi da ƙananan shigar / share maye gurbi na iya haifar da asarar ayyukan ƙwayar ƙwayar cuta yayin da ke shafar mahimman wuraren aiki. Bambance-bambancen tsarin da ke haifar da asarar ayyukan ƙwayoyin cuta masu hana ƙari sun haɗa da ɓarna ko cikakken shafewar kwayoyin halitta da sauran bambance-bambancen kwayoyin halitta waɗanda ke haifar da lalata tsarin karatun kwayoyin halitta. Ƙananan bambance-bambancen da ke haifar da ingantacciyar aikin oncogenes sun haɗa da maye gurbi na rashin kuskure da shigarwa / gogewa na lokaci-lokaci wanda ke yin niyya ga mahimman wuraren aikin furotin. A lokuta da ba kasafai ba, raguwar furotin ko maye gurbi na iya haifar da kunna oncogenes. Bambance-bambancen tsarin da ke haifar da kunna oncogene sun haɗa da haɗakar halitta, shafewar kwayoyin halitta, da kwafi.
Fassarar asibiti na bambancin kwayoyin halitta tana tantance mahimmancin asibiti na maye gurbi da aka gano, watau yuwuwar binciken su, tsinkaye, ko ƙimar warkewa. Akwai tsare-tsaren ƙididdiga na tushen shaida da yawa waɗanda za a iya amfani da su don jagorantar fassarar asibiti na bambancin genomic.
Cibiyar Nazarin Ciwon Kankara ta Memorial Sloan-Kettering ta Mahimman Bayanan Magungunan Oncology (OncoKB) tana rarraba bambance-bambancen kwayoyin halitta zuwa matakai hudu dangane da ƙimar tsinkayar su don amfani da miyagun ƙwayoyi: Level 1/2, FDA-an yarda, ko kuma daidaitattun kwayoyin halitta waɗanda ke tsinkayar amsa ta takamaiman nuni ga magani da aka yarda; Mataki na 3, FDA-an yarda ko masu ba da izini na biomarkers waɗanda ke yin la'akari da martani ga magungunan da aka yi niyya da suka nuna alƙawarin a cikin gwaje-gwajen asibiti, da Level 4, waɗanda ba FDA-amince da masu ilimin halitta waɗanda ke yin hasashen amsa ga magungunan da aka yi niyya na labari waɗanda suka nuna tabbataccen shaidar ilimin halitta a cikin gwaji na asibiti. An ƙara ƙaramin rukuni na biyar da ke da alaƙa da juriya na jiyya.
Al'umman Amurkawa don kwayar cutar kwayoyin halitta (amp) / Aboki na Clincology na Clincology (ASCO) / Kwalejin Amurkawa na Amurka (Colleal of Colleologists na SPOMIC (aji) / sa ni, tare da karfin asibiti; Mataki na II, tare da yuwuwar mahimmancin asibiti; Mataki na III, ba a san mahimmancin asibiti ba; Grade IV, ba a san yana da mahimmancin asibiti ba. Bambance-banbancen digiri na I da na II ne kawai ke da kima don yanke shawarar magani.
ESMO's Molecular Target Clinical Operability Scale (ESCAT) yana rarraba bambance-bambancen kwayoyin halitta zuwa matakai shida: Mataki na I, maƙasudin da suka dace da amfani na yau da kullun; Mataki na II, makasudin da har yanzu ana nazarin, yana yiwuwa a yi amfani da shi don tantance yawan majinyata da za su iya amfana daga maganin da aka yi niyya, amma ana buƙatar ƙarin bayanai don tallafawa. Mataki na III, bambance-bambancen jinsin da aka yi niyya waɗanda suka nuna fa'idar asibiti a cikin wasu nau'in ciwon daji; Darajoji na IV, bambance-bambancen jinsin da aka yi niyya kawai wanda ke goyan bayan hujjoji na asali; A cikin digiri na V, akwai shaidar da za ta goyi bayan mahimmancin asibiti na yin niyya ga maye gurbin, amma maganin miyagun ƙwayoyi guda ɗaya a kan manufa ba ya tsawanta rayuwa, ko kuma za'a iya amfani da dabarun jiyya; Grade X, rashin darajar asibiti.