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Yi rijista da takamaiman bayananka da takamaiman maganin da kake sha'awa kuma za mu daidaita bayanan da ka bayar tare da labarai a cikin babban rumbun adana bayanai kuma nan take za mu aiko maka da kwafin PDF ta imel.
Stella S, Vitale SR, Martorana F, Massimino M, Pavone G, Lanzafame K, Bianca S, Barone C, Gorgone C, Fichera M, Manzella L
Stefania Stella, 1,2 Silvia Rita Vitale, 1,2 Federica Martorana, 1,2 Michele Massimino, 1,2 Giuliana Pavone, 3 Katia Lanzafame, 3 Sebastiano Bianca, 4 Chiara Barone, 5 Cristina Gorgone, 6 Marco Fichera, 12 Sashen Liviazella 1 da Clinical Manuniya Magunguna, Jami'ar Catania, Catania, 95123, Italiya; 2 Cibiyar Gwajin Oncology da Hematology, AOU Policlinico "G.Rodolico - San Marco", Catania, 95123, Italiya; 3 Medical Oncology, AOU Policlinico "G. Rodolico - San Marco", Catania, 95123, Italiya; 4 Medical Genetics, ARNAS Garibaldi, Catania, 95123, Italiya; 5 Magungunan Halittar Jini, ASP, Syracuse, 96100, Italiya; 6 Sashen Kimiyyar Halittu da Fasahar Halittu, Jami'ar Catania, Ilimin Halittar Jini na Likitanci, Catania, Italiya, 95123; 7Oasi Research Institute-IRCCS, Troina, 94018, Italiya Sadarwa: Stefania Stella, tel +39 095 378 1946, imel [email protected]; [email protected] Manufa: Sauye-sauyen ƙwayoyin cuta a cikin BRCA1 da BRCA2 da kuma ciwon nono da aka kafa (BC), ovary (OC) da sauran waɗanda ke da alaƙa da haɗarin ciwon daji na rayuwa. Gwaji don kwayar halittar BRCA shine mabuɗin tantance haɗarin mutum ɗaya, da kuma nemo hanyoyin rigakafi a cikin masu ɗauke da cutar da kuma daidaita jiyya ga marasa lafiya da cutar kansa. Yaɗuwar canje-canjen BRCA1 da BRCA2 ya bambanta sosai a yankuna daban-daban, kuma kodayake akwai bayanai kan bambance-bambancen cututtukan BRCA a cikin iyalan Sicilian, nazarin da aka yi niyya musamman ga al'ummomi a gabashin Sicily ba su da shi. Manufar bincikenmu ita ce bincika faruwar da rarrabawar canje-canjen ƙwayoyin cuta na BRCA a cikin ƙungiyar marasa lafiya na BC daga gabashin Sicily da kuma tantance alaƙarsu da takamaiman halayen BC ta amfani da jerin tsararraki na gaba. Kasancewar canje-canjen da ke da alaƙa da matakin ƙari da ma'aunin yaduwa. SAKAMAKO: Gabaɗaya, marasa lafiya 35 (9%) suna da bambancin cututtukan BRCA, 17 (49%) a cikin BRCA1 da 18 (51%) a cikin BRCA2. Sauye-sauyen BRCA1 sun fi yawa a cikin marasa lafiya na BC marasa lafiya marasa lafiya marasa lafiya uku, yayin da maye gurbin BRCA2 ya fi yawa a cikin haske BC marasa lafiya. Idan aka kwatanta da waɗanda ba sa ɗauke da cutar, waɗanda ke da nau'ikan BRCA1 suna da babban matakin ciwon daji da kuma yawan yaduwar cutar. Kammalawa: Bincikenmu ya ba da taƙaitaccen bayani game da matsayin maye gurbin BRCA a cikin marasa lafiya na BC daga gabashin Sicily kuma ya tabbatar da rawar da binciken NGS ke takawa wajen gano marasa lafiya da ke ɗauke da cutar BC ta gado. Gabaɗaya, waɗannan bayanan sun yi daidai da shaidun da suka gabata waɗanda ke goyan bayan gwajin BRCA don ingantaccen rigakafi da maganin cutar kansa a cikin masu ɗauke da cutar.
Ciwon daji na mama (BC) shine cutar da ta fi yawa a duniya kuma mafi muni a cikin mata.1 An yi nazari sosai kan siffofin halittu waɗanda ke tantance hasashen BC da halayen asibiti a tsawon lokaci kuma an yi bayani dalla-dalla a kansu. A gaskiya ma, ana amfani da wasu alamomin maye gurbin BC a halin yanzu don rarraba BC zuwa nau'ikan kwayoyin halitta daban-daban. Su ne estrogen (ER) da/ko progesterone receptor (PgR), haɓakar epidermal growth factor receptor 2 (HER2), ƙimar yaduwa Ki-67 da matakin ƙari (G).2 Haɗin waɗannan masu canji sun gano waɗannan rukunan BC: 1) Ciwon daji na Luminal, wanda ke nuna bayyanar ER da/ko PgR, ya kai kashi 75% na BCs. An ƙara raba waɗannan ciwace-ciwacen zuwa Luminal A, lokacin da Ki-67 ya kasance ƙasa da 20% da HER2 mara kyau, da Luminal B, lokacin da Ki-67 ya kasance daidai ko sama da 20% kuma a gaban haɓakar HER2, ba tare da la'akari da ƙimar yaduwa ba; 2) Ciwon daji na HER2+ waɗanda ba su da ER da PgR amma suna nuna ƙaruwar HER2. Wannan rukuni ya ƙunshi kashi 10% na dukkan ciwon daji na nono; 3) Ciwon daji na nono mai rashin sakamako uku (TNBC), wanda ba ya nuna bayyanar ER da PgR da ƙaruwar HER2, ya kai kusan kashi 15% na ciwon daji na nono.2-4
Daga cikin waɗannan nau'ikan BC, matakin ciwon daji da kuma yawansa suna wakiltar alamun cututtuka na giciye waɗanda ke da alaƙa kai tsaye da kuma kai tsaye da ƙarfin ciwon daji da kuma hasashen ci gaba.5,6
Baya ga siffofin halittu da aka ambata a baya, rawar da sauye-sauyen kwayoyin halitta da aka gada suka haifar da ci gaban BC ya zama muhimmi a cikin 'yan shekarun nan.7 Kimanin 1 cikin 10 na ciwace-ciwacen nono ana gadonsu ne saboda sauye-sauyen kwayoyin halitta a cikin takamaiman kwayoyin halitta.8 Manyan bincike guda biyu na cututtuka da suka shafi mata sama da 180,000 sun gano wani rukuni na kwayoyin halitta takwas (watau ATM, BARD1, BRCA1, BRCA2, CHK2, PALB2, RAD51C, da RAD51D) wadanda ke da alhakin BC na gado. Daga cikin wadannan kwayoyin halitta, BRCA1 da BRCA2 (wanda daga baya ake kira BRCA1/2) sun nuna mafi karfin alaka da ciwace-ciwacen nono.9-12 A gaskiya ma, maye gurbi na BRCA1/2 na germline yana kara yawan hadarin BC na tsawon rai da sauran cututtuka, gami da ovarian, prostate, pancreas, colorectal, da melanoma. Daga shekaru 13 zuwa 80, yawan kamuwa da BC shine 72% a cikin mata masu bambancin BRCA1 pathogenic variant (PV) da 69% a cikin mata masu BRCA2 PV.14
Abin lura shi ne, wani littafi da aka buga kwanan nan ya nuna cewa haɗarin BC ya dogara ne da nau'in PV. A gaskiya ma, idan aka kwatanta da bambance-bambancen da ke rage cututtuka, bambance-bambancen rashin fahimta, musamman a cikin kwayar halittar BRCA1, suna da alaƙa da raguwar haɗarin BC, musamman a cikin tsofaffi mata.15
Kasancewar BRCA1 ko BRCA2 PV yana da alaƙa da siffofi daban-daban na halitta da na asibiti.16,17 BCs masu alaƙa da BRCA1 suna da saurin kamuwa da cuta a asibiti, ba su da bambanci sosai, kuma suna da saurin yaduwa. Waɗannan ciwace-ciwacen yawanci suna da alamun rashin lafiya sau uku kuma suna da farkon farkon farawa. Ciwace-ciwacen da ke faruwa a cikin marasa lafiya da aka canza BRCA2 galibi suna nuna matsakaicin matsayi zuwa bambance-bambancen da aka bambanta da kuma alamun yaduwa masu canzawa. Waɗannan ciwace-ciwacen sun fi yawa a cikin lumen B kuma yawanci suna faruwa a cikin tsofaffi.16-18 Abin lura, maye gurbi a cikin BRCA1 da BRCA2 yana ƙara yawan jin daɗin jiyya, gami da gishirin platinum da magunguna da aka yi niyya kamar poly(ADP-ribose) polymerase inhibitors (PARPi).19,20
A cikin 'yan shekarun da suka gabata, aiwatar da tsarin tsara na gaba (NGS) a cikin aikin asibiti ya ba da damar ƙara yawan marasa lafiya na BC su yi gwajin kwayoyin halitta don cututtukan da ke iya kamuwa da cutar kansa, gami da BRCA1/2.21 A lokaci guda, ma'anoni bisa ga takamaiman sharuɗɗa game da tarihin iyali, yanayin alƙaluma, da halayen asibiti don gano mutanen da suka cancanci gwajin BRCA1/2.22,23 A cikin wannan mahallin, shaidu suna taruwa akan gwajin BRCA1/2 a cikin takamaiman al'ummomi, suna nuna bambance-bambance a cikin yankuna na yanki.24-27 Kodayake akwai rahotanni game da ƙungiyar BC a yammacin Sicily, ƙarancin bayanai da ake samu akan gwajin BRCA1/2 a cikin al'ummar gabashin Sicily.28,29
A nan mun bayyana sakamakon gwajin germline BRCA1/2 a cikin marasa lafiya na BC daga gabashin Sicily, wanda ke ƙara alaƙa da kasancewar maye gurbi na BRCA1 ko BRCA2 tare da manyan fasalulluka na cututtukan cututtukan ƙwayoyin cuta na waɗannan ƙari.
An gudanar da wani bincike na baya-bayan nan a "Cibiyar Nazarin Cututtukan Jijiyoyi da Hematology" a Asibitin Policlinico. Rodolico - San Marco a Catania. Daga Janairu 2017 zuwa Maris 2021, an tura jimillar marasa lafiya 455 da ke fama da cutar kansar nono da ta mahaifa, melanoma, pancreas ko prostate zuwa dakin gwaje-gwajen kwayoyin halittarmu don gwajin kwayoyin halitta na BRCA/2. An gudanar da wannan binciken ne bisa ga Sanarwar Helsinki, kuma duk mahalarta sun ba da izini a rubuce kafin a yi nazarin kwayoyin halitta.
An tantance halayen tarihi da na halitta (ER, PgR, matsayin HER2, Ki-67, da kuma matakin) na BC akan samfuran biopsy na asali ko na tiyata, idan aka yi la'akari da abubuwan da ke haifar da ƙari masu ƙarfi kawai. Dangane da waɗannan halaye, an rarraba BCs kamar haka: luminal A (ER+ da/ko PgR+, HER2-, Ki-67 <20%), luminal B (ER+ da/ko PgR+, HER2-, Ki-67≥20%), luminal B-HER2+ (ER da/ko PgR+, HER2+), HER2+ (ER da PgR-, HER2+) ko uku negative (ER da PgR-, HER2-).
Kafin a tantance matsayin maye gurbin BRCA1 da BRCA2, wata ƙungiya mai fannoni daban-daban, ciki har da likitan kansa, masanin kwayoyin halitta, da kuma masanin ilimin halayyar ɗan adam, ta gudanar da shawarwari kan kwayoyin halittar ƙari ga kowane majiyyaci don tantance kasancewar BRCA1 da/ko BRCA1. ko mutanen da ke da babban haɗarin kamuwa da cutar PV a cikin kwayar halittar BRCA2. An gudanar da zaɓin marasa lafiya bisa ga jagororin Ƙungiyar Magungunan Lafiya ta Italiya (AIOM) da shawarwarin Sicilian na gida.30,31 Waɗannan sharuɗɗan sun haɗa da: (i) tarihin iyali na bambance-bambancen cututtuka da aka sani a cikin kwayoyin halittar da ke da saurin kamuwa da cutar (misali, BRCA1, BRCA2, TP53, PTEN); (ii) maza da ke da BC; (iii) waɗanda ke da BC da OC; (iv) mata da ke da BC <36 shekaru, TNBC <60 shekaru, ko BC na biyu <50 shekaru; (v) tarihin lafiyar mutum na BC <50 shekaru da aƙalla dangi ɗaya na farko: (a) BC <50 shekaru; (b) OC mara mucinous da mara iyaka na kowane zamani; (c) BC na biyu; (d) BC na namiji; (e) ciwon daji na pancreas; (f) ciwon daji na prostate; (vi) tarihin mutum biyu ko fiye na BC > shekaru 50 da tarihin iyali na BC, OC, ko ciwon daji na pancreas ga dangi waɗanda suke dangi na farko ga juna (gami da dangin da take dangi na farko); (vii) Tarihin mutum na OC da aƙalla dangi ɗaya na farko: (a) BC <50 shekaru; (b) NOC; (c) BC na biyu; (d) namiji BC; (vii) mace mai OC mai babban matakin serous.
An samo samfurin jinin da ke kewaye da mutum 20 mL daga kowane majiyyaci kuma aka tattara shi a cikin bututun EDTA (BD Biosciences). An ware DNA na genomic daga samfuran jini na 0.7 mL ta amfani da Kit ɗin Rage Kitse na QIAsymphony DSP DNA Midi (QIAGEN, Hilden, Italiya) bisa ga umarnin masana'anta kuma an wuce ta cikin na'urar auna Fluorometer ta Qubit® 3.0 (Thermo Fisher Scientific, Waltham, MA, Amurka). Ana yin amfani da Oncomine™ BRCA Research Assay Chef, wanda aka shirya don a ɗora shi cikin Ion AmpliSeq™ Chef Reagents DL8 Kit don shirya ɗakin karatu ta atomatik bisa ga umarnin masana'anta. Kayan aikin ya ƙunshi wuraren farawa guda biyu na PCR waɗanda za a iya amfani da su don nazarin dukkan kwayoyin halittar BRCA1 (NM_007300.3) da BRCA2 (NM_000059.3). A takaice, an ƙara 15 µL na kowane samfurin DNA mai narkewa (10 ng) a cikin faranti masu barcode don shirya ɗakin karatu kuma an ɗora duk abubuwan haɗin gwiwa da abubuwan amfani akan kayan aikin Ion Chef™. Daga nan aka yi shirye-shiryen ɗakin karatu ta atomatik da tattara samfuran ɗakin karatu mai barcode akan kayan aikin Ion Chef™. Sannan an tantance adadin ɗakunan karatu da aka shirya ta amfani da Qubit® 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, Amurka) bisa ga umarnin masana'anta. A ƙarshe, ana haɗa ɗakunan karatu a cikin rabon equimolar a cikin bututun samfurin ɗakin karatu na Ion Chef™ (wanda aka sanya lambar barcode) An yi amfani da na'urar Ion Torrent S5 (Thermo Fisher Scientific) (Thermo Fisher Scientific) wajen yin jerin gwanon bayanai (sequencing). An yi amfani da na'urar Ion Torrent S5 (Thermo Fisher Scientific) wajen amfani da na'urar Ion 510 Chip (Thermo Fisher Scientific). An yi nazarin bayanai ta hanyar Amplicon Suite (SmartSeq srl) da kuma Ion Reporter Software.
Duk sunayen bambance-bambancen sun bi ka'idojin yanzu na Ƙungiyar Bambancin Halittar Dan Adam, wanda ake samu akan layi (HGVS, http://www.hgvs.org/mutnomen). An bayyana mahimmancin asibiti na bambance-bambancen BRCA1/2 ta amfani da rarrabuwar International Consortium ENIGMA (Cibiyar Bayar da Shaida don Fassarar Germline Mutant Alleles, https://enigmaconsortium.org/) da kuma tuntuɓar bayanai daban-daban kamar ARUP, BRCAEXCHANGE, ClinVar, IARC_LOVD, da UMD. Rarrabawar ta haɗa da nau'ikan haɗari guda biyar daban-daban: marasa lahani (rukuni na I), mai yiwuwa marasa lahani (rukuni na II), bambancin mahimmancin da ba a tabbatar ba (VUS, rukuni na III), mai yuwuwar masu cutarwa (rukuni na IV), da masu cutarwa (rukuni na V). VarSome sun kuma yi nazarin tasirin maye gurbi akan tsarin furotin da aiki, kayan aiki mai ba da labari tare da damar shiga bayanan bayanai 30.32
Domin sanya mahimmancin asibiti ga kowace VUS, an yi amfani da waɗannan algorithms na hasashen furotin na lissafi: MUTATION TASTER, 33 PROVEAN-SIFT (http://provean.jcvi.org/index.php), POLYPHEN-2 (http:///genetics.bwh.harvard.edu/pph2/) da Align-GVGD (http://agvgd.hci.utah.edu/agvgd_input.php). An ɗauki nau'ikan da aka rarraba a matsayin aji na 1 da na 2 a matsayin nau'in daji.
Tsarin Sanger ya tabbatar da kasancewar kowace bambance-bambancen cuta. A takaice, an tsara wasu takamaiman firam guda biyu don kowane bambance-bambancen da aka gano ta hanyar amfani da jerin sunayen kwayoyin halittar BRCA1 da BRCA2 (NG_005905.2, NM_007294.3 da NG_012772.3, NM_000059.3, bi da bi). Saboda haka, an yi PCR da aka yi niyya sannan aka bi Sanger sequencing.
An gwada marasa lafiya da aka gwada ba su da kwayar halittar BRCA1/2 ta hanyar amfani da multiplex ligation-dependent probe amplification (MLPA) bisa ga umarnin masana'anta don tantance kasancewar manyan sake fasalin kwayoyin halitta (LGR). A takaice, ana cire samfuran DNA kuma ana amfani da har zuwa 60 na BRCA1 da BRCA2 na musamman na kwayoyin halitta, kowannensu yana gano takamaiman jerin DNA kimanin nucleotides 60 a tsayi. Sannan an yi nazarin samfuran ƙara girman PCR, waɗanda suka ƙunshi saitin musamman na amplicons, ta hanyar amfani da capillary electrophoresis da kuma software na Cofalyser.Net tare da teburin Cofalyser na musamman na rukuni (www.mrcholland.com).
An haɗa zaɓaɓɓun masu canjin yanayin asibiti (matakin histological da ma'aunin yaduwar Ki-67%) da kasancewar BRCA1/2 PV, waɗanda aka ƙididdige ta amfani da software na Prism v. 8.4 ta amfani da gwajin Fisher daidai wanda ake ɗauka cewa ƙimar p <0.05 tana da mahimmanci.
Tsakanin Janairu 2017 da Maris 2021, an duba marasa lafiya 455 don gano maye gurbi na BRCA1/2 na germline. An gudanar da gwajin maye gurbi a Cibiyar Nazarin Cututtukan Jiki da Hematology ta Asibitin Policlinico. Dangane da jagorar Sicilian (http://www.gurs.regione.sicilia.it/Indicep1.htm, N. 02-Venerdì 10 Gennaio 2020), jimillar marasa lafiya 389. Akwai ciwon nono, ciwon daji na ovarian 37, ciwon daji na pancreas 16, ciwon daji na prostate 8 da melanoma 5. An nuna rarrabawar marasa lafiya bisa ga nau'in ciwon daji da sakamakon bincike a Hoto na 1.
Hoto na 1 ya nuna jadawalin kwararar da ke nuna taƙaitaccen bayani game da binciken. An gwada marasa lafiya da ke da ciwon nono, melanoma, pancreas, prostate, ko ovarian don maye gurbi a cikin kwayoyin halittar BRCA1 da BRCA2.
Takaitattun bayanai: PVs, bambancin cututtuka; VUS, bambancin mahimmancin da ba a tabbatar ba; WT, jerin BRCA1/2 na daji.
Mun mayar da hankali sosai kan bincikenmu kan rukunin masu fama da cutar kansar mama. Marasa lafiya suna da matsakaicin shekaru na shekaru 49 (tsakanin 23-89) kuma galibi mata ne (n=376, ko 97%).
Daga cikin waɗannan mutane, 64 (17%) sun sami maye gurbi na BRCA1/2 kuma duk mata ne. Talatin da biyar (9%) suna da PV kuma 29 (7.5%) suna da VUS. Goma sha bakwai (48.6%) daga cikin bambance-bambancen cuta guda 35 sun faru a cikin BRCA1 da 18 (51.4%) a cikin BRCA2, yayin da VUS 5 ya faru a cikin BRCA1 (17.2%) da 24 (82.8%) a cikin BRCA2 (Hoto na 1 da 2). LGR bai kasance a cikin nazarin MLPA ba.
Hoto na 2. Binciken maye gurbi na BRCA1 da BRCA2 a cikin marasa lafiya 389 na ciwon nono.(A) Rarraba bambance-bambancen cututtuka (PV) (ja), bambance-bambancen mahimmancin da ba a tabbatar ba (VUS) (orange), da WT (shuɗi) a cikin marasa lafiya 389 na ciwon nono; (B) marasa lafiya 389 na ciwon nono suna da bambance-bambancen cututtuka na BRCA1/2 (PVs). Daga cikinsu, 17 (48.6%) sune masu ɗauke da PV na BRCA1 (ja mai duhu) kuma 18 (51.4%) sune masu ɗauke da BRCA2 (ja mai haske); (C) 29 (7.5%) na mutane 389 suna ɗauke da VUS, 5 (17.2%) kwayoyin halittar BRCA1 (orange mai duhu) da 24 (82.8%) kwayoyin halittar BRCA2 (orange mai haske).
Takaitattun bayanai: PVs, bambancin cututtuka; VUS, bambancin mahimmancin da ba a tabbatar ba; WT, jerin BRCA1/2 na daji.
Mun sake bincika yawan nau'ikan ƙwayoyin BC a cikin marasa lafiya da ke ɗauke da BRCA1/2 PV. Rarrabawar ta haɗa da marasa lafiya 2 (5.7%) masu ɗauke da ƙwayayen A, 15 (42.9%) masu ɗauke da ƙwayayen B, 3 (8.6%) masu ɗauke da ƙwayayen B-HER2+, 2 (5.7%) masu ɗauke da ƙwayayen HER2+ da 13 (37.1%) masu ɗauke da TNBC. Daga cikin marasa lafiya da ke ɗauke da ƙwayayen BRCA1, 5 (29.4%) suna ɗauke da ƙwayayen B BC, 2 (11.8%) suna ɗauke da cutar HER2+, kuma 10 (58.8%) suna ɗauke da TNBC. Ciwon da ba tare da maye gurbin BRCA1 ba ko dai masu ɗauke da ƙwayayen A ne ko masu ɗauke da ƙwayayen B-HER2+ (Hoto na 3). A cikin ƙungiyar BRCA2 masu ɗauke da ƙwayayen, ciwon 10 (55.6%) masu ɗauke da ƙwayayen B ne masu ɗauke da ƙwayayen B, 3 (16.7%) masu ɗauke da ƙwayayen B-HER2+ ne masu ɗauke da ƙwayayen B, 3 (16.7%) masu ɗauke da ƙwayayen TNBC da 2 (11.1%) masu ɗauke da ƙwayayen A ne masu ɗauke da ƙwayayen A. (Hoto na 3). Babu ciwon HER2+ a cikin wannan rukunin. Don haka, maye gurbi na BRCA1 ya zama ruwan dare a cikin marasa lafiya na TNBC, yayin da canje-canje na BRCA2 sun fi yawa a cikin mutanen da ke da lumen B.
Hoto na 3 Yaɗuwar ƙananan nau'ikan ciwon nono a cikin marasa lafiya da ke da bambance-bambancen cututtuka a cikin BRCA1 da BRCA2. Histograms da ke nuna rarrabawar BRCA1- (ja mai duhu) da BRCA2- (ja mai haske) PVs tsakanin ƙananan nau'ikan ƙwayoyin cuta na marasa lafiya da ke da ciwon nono. Lambobin da aka ruwaito a cikin kowane akwati suna wakiltar kaso na marasa lafiya da ke da BRCA1 da BRCA2 PV ga kowane nau'in ciwon nono.
Takaitattun bayanai: PVs, nau'in cutarwa; HER2+, mai karɓar haɓakar epidermal na ɗan adam 2 mai kyau; TNBC, ciwon nono mai kama da na uku.
Bayan haka, mun tantance nau'in da kuma inda kwayar halittar BRCA1 da BRCA2 PVs take. A cikin BRCA1 PV, mun lura da bambance-bambancen nucleotide guda 7 (SNVs), gogewa 6, kwafi 3 da sakawa 1. Sauye-sauye guda ɗaya kawai (c.5522delG) yana wakiltar sabon bincike. Mafi yawan BRCA1 PV da aka gano a cikin duka biyun shine c.5035_5039delCTAAT. Wannan sauyi ya ƙunshi share nucleotides guda biyar (CTAAT) a cikin BRCA1 exon 15, wanda ya haifar da maye gurbin amino acid leucine da tyrosine a codon 1679, kuma saboda canjin tsarin fassara tare da hasashen madadin dakatarwar codon yana haifar da yanke furotin da wuri. Duk sauran canje-canje ana gano su a cikin yanayi ɗaya kawai. Abin lura shine, ɗaya daga cikin PVs da aka ruwaito yana cikin yankin yarjejeniya na wurin raba (c.4357+1G>T) (Tebur 1).
Dangane da BRCA2 PV, mun lura da gogewa sau 6, SNV guda 6 da kwafi 2. Babu ɗaya daga cikin canje-canjen da aka samu da ba sabon abu ba ne. Sauye-sauye uku sun sake faruwa a cikin al'ummarmu, c.428dup da c.8487+1G>A da aka lura a cikin mutane 3, sai kuma c.5851_5854delAGTT da aka samo a cikin lokuta biyu. Sauye-sauyen c.428dup ya ƙunshi maimaita C a cikin exon 5 na BRCA2, wanda aka annabta zai ƙunshi furotin da aka yanke, wanda ba shi da aiki. Sauye-sauyen c.8487+1G>Cututtuka suna faruwa a yankin intronic na BRCA2 intron 19 (± 1,2) kuma yana shafar jerin haɗin gwiwa, wanda ke haifar da canjin haɗin gwiwa wanda ke haifar da rashin daidaituwa ko rashin furotin. Bambancin cutar c.5851_5854delAGTT ya faru ne saboda gogewar 4-nucleotide daga matsayin nucleotide 5851 zuwa 5854 a cikin lambar exon 10 na kwayar halittar BRCA2 da yana haifar da canjin tsari tare da annabta madadin dakatarwar codon (p.S1951WfsTer). Abin lura, kamar yadda aka ruwaito a baya, an gano duka canje-canje c.631G>A da c.7008-2A>T a cikin majiyyaci ɗaya.34 Sauyin farko ya ƙunshi maye gurbin adenosine (A) a cikin BRCA2 exon 7 tare da guanine (G) wanda ke ɗauke da nucleotide wanda ke haifar da canjin valine zuwa isoleucine a codon 211, isoleucine Amino acid amino acid ne mai halaye masu kama da juna. Wannan canjin yana shafar haɗin mRNA na yau da kullun. Bambancin na biyu yana cikin yankin intronic kuma yana haifar da maye gurbin A zuwa thymine (T) sau biyu kafin exon 13 na kwayar halittar da ke ƙunshe da BRCA2. Canjin c.7008-2A>T na iya samar da rubuce-rubuce da yawa na tsayi daban-daban. Bugu da ƙari, a cikin ƙungiyar BRCA2 PVs, 4 daga cikin canje-canje 18 (22.2%) sun kasance intronic.
Sai muka zana taswirar maye gurbi mai illa na BRCA1/2 a yankunan aiki da yankunan da ke ɗaure furotin (Hoto na 4). A cikin kwayar halittar BRCA1, kashi 50% na PVs suna cikin yankin ƙungiyar ciwon nono (BCCR), yayin da kashi 22% na maye gurbi suna cikin yankin ƙungiyar ciwon daji na ovarian (OCCR) (Hoto na 4A). A cikin BRCA2 PV, kashi 35.7% na bambance-bambancen suna cikin yankin BCCR kuma kashi 42.8% na maye gurbi suna cikin OCCR (Hoto na 4B). Na gaba, mun tantance wurin da PV yake a cikin yankunan furotin na BRCA1 da BRCA2. Ga furotin na BRCA1, mun sami PVs guda uku a cikin madauki da yankunan coil mai coil, da kuma maye gurbi guda biyu a cikin yankin BRCT (Hoto na 4A). Ga furotin na BRCA2, an zana PVs guda 4 zuwa yankin maimaita BRC, yayin da aka gano canje-canje na intronic da 3 na exonic a cikin yankunan oligo/oligosaccharide-binding (OB) da hasumiya (T) (Hoto na 4B). 4B).
Hoto na 4. Wakiltar tsari na sunadaran BRCA1 da BRCA2 da kuma inda aka samo bambance-bambancen cututtuka. Wannan adadi yana nuna rarraba bambance-bambancen cututtuka na BRCA1 (A) da BRCA2 (B) a cikin marasa lafiya da ke fama da cutar kansar nono. An nuna maye gurbi na waje a cikin shuɗi, yayin da aka nuna bambance-bambancen intronic a cikin orange. Tsawon sandar yana wakiltar adadin shari'o'in. An ruwaito sunadaran BRCA1 da BRCA2 da yankunan aikinsu. (A) Sunadaran BRCA1 sun ƙunshi yankin madauki (RING) da jerin wuraren da ake amfani da su a cikin nukiliya (NLS), yankin coiled-coil, yankin SQ/TQ cluster (SCD), da yankin BRCA1 C-terminal (BRCT).(B) Sunadaran BRCA2 sun ƙunshi maimaita BRC guda takwas, yankin da ke ɗaure DNA tare da yankin helical (Helical), ninka oligonucleotide/oligosaccharide-binding (OB) guda uku, yankin hasumiya (T), da An NLS a gefen C. An nuna yankunan da ake kira Yankin Cluster Cancer Region (BCCR) da Ovarian Cancer Cluster Region (OCCR) a ƙasa.*Yana wakiltar sauye-sauyen da ke ƙayyade dakatarwar codons.
Sai muka binciki siffofin cutar BC da ke iya alaƙa da kasancewar BRCA1/2 PV. An sami cikakkun bayanan asibiti ga marasa lafiya 181 na BRCA1/2 marasa lafiya (marasa ɗauke da cutar) da duk masu ɗauke da cutar (n = 35). Akwai alaƙa tsakanin ƙimar yaduwar cutar da kuma matakinta.
Mun ƙididdige rarrabawar Ki-67 bisa ga matsakaicin ƙungiyarmu (25%, kewayon <10-90%). An ayyana waɗanda ke da Ki-67 <25% a matsayin "ƙananan Ki-67", yayin da mutanen da ke da ƙimar ≥ 25% an ɗauke su a matsayin "babban Ki-67". An gano bambance-bambance masu mahimmanci na Ki-67 (p<0.01) tsakanin waɗanda ba sa ɗaukar kaya da kuma masu ɗaukar PV na BRCA1 (Hoto na 5A).
Hoto na 5 Alaƙar Ki-67 da rarrabawar maki a cikin mata masu fama da cutar kansar mama tare da kuma ba tare da BRCA1 da BRCA2 PVs ba. (A) Akwatin zane yana nuna matsakaicin ƙimar Ki-67 a cikin marasa lafiya 181 marasa lafiya na BC waɗanda ba sa ɗauke da cutar idan aka kwatanta da marasa lafiya na BRCA1 (18) ko BRCA2 (17). An yi la'akari da ƙimar P da ke ƙasa da 0.5 a matsayin mahimmanci a kididdiga. (B) Histogram wanda ke wakiltar sanya marasa lafiya na BC ciwon daji cikin ƙungiyoyin matakin histological (G2 da G3) bisa ga matsayin maye gurbin BRCA1 da BRCA2 (masu ɗauke da cutar WT, masu ɗauke da BRCA1 da BRCA2 PVs).
Haka kuma, mun bincika ko matakin ciwon daji yana da alaƙa da kasancewar BRCA1/2 PV. Tunda G1 BC ba ya nan a cikin al'ummarmu, mun raba marasa lafiya zuwa ƙungiyoyi biyu (G2 ko G3). Dangane da sakamakon Ki-67, binciken ya nuna alaƙa mai mahimmanci tsakanin matakin ciwon daji da maye gurbin BRCA1, tare da mafi girman rabo na ciwon daji na G3 a cikin masu ɗauke da BRCA1 idan aka kwatanta da waɗanda ba sa ɗauke da cutar (p<0.005) (Hoto na 5B).
Ci gaba a fasahar tsara kwayoyin halitta ta DNA ya ba da damar ci gaba mara misaltuwa a gwajin kwayoyin halitta na BRCA1/2, tare da muhimman tasiri ga marasa lafiya da ke da tarihin cutar kansa a cikin iyali. Zuwa yanzu, an gano kuma an rarraba nau'ikan BRCA1/2 kusan 20,000 bisa ga Ƙungiyar Nazarin Kwayoyin Halitta ta Amurka 35 da tsarin ENIGMA.35,36 An san cewa bambancin BRCA1/2 ya bambanta sosai a yankuna daban-daban.37 A cikin Italiya, ƙimar BRCA1/2 PVs ya kasance daga 8% zuwa 37%, yana nuna bambancin da ke tsakanin ƙasashe.38,39 Tare da yawan jama'a kusan miliyan 5, Sicily ita ce yanki na biyar mafi girma a Italiya dangane da adadin mazauna. Kodayake akwai bayanai kan rarraba BRCA1/2 a yammacin Sicily, babu wata shaida mai zurfi a gabashin tsibirin.
Bincikenmu yana ɗaya daga cikin rahotannin farko kan yawan kamuwa da cutar BRCA1/2 PV a cikin marasa lafiya na BC a gabashin Sicily.28 Mun mayar da hankali kan nazarinmu kan BC, domin wannan ita ce cutar da ta fi yawa a cikin ƙungiyarmu.
Lokacin da aka gwada marasa lafiya 389 na BC, kashi 9% suna ɗauke da BRCA1/2 PVs, waɗanda aka rarraba daidai tsakanin BRCA1 da BRCA2. Waɗannan sakamakon sun yi daidai da waɗanda aka ruwaito a baya a cikin al'ummar Italiya.28 Abin sha'awa, kashi 3% (13/389) na ƙungiyarmu maza ne. Wannan adadin ya fi yadda ake tsammani ga ciwon nono na maza (1% na dukkan BCs),40 yana nuna zaɓin al'ummominmu bisa ga haɗarin maye gurbi na BRCA1/2. Duk da haka, babu ɗayan waɗannan mazan da ya haifar da BRCA1/2 PV, don haka sun kasance 'yan takara don ƙarin nazarin kwayoyin halitta don kawar da kasancewar maye gurbi marasa yawa kamar PALB2, RAD51C da D, da sauransu. An samo bambance-bambancen da ba su da tabbas a cikin kashi 7% na batutuwa waɗanda BRCA2 VUS ya bayyana. Ko da wannan sakamakon ya yi daidai da shaidar da ta riga ta kasance.28,41,42
Lokacin da muka yi nazarin rarraba nau'ikan kwayoyin BC a cikin mata masu maye gurbin BRCA1/2, mun tabbatar da alaƙar da aka sani tsakanin TNBC da BRCA1 PV (58.8%) da kuma tsakanin haske B BC da BRCA2 PV (55.6%).16,43 Ciwon daji na haske A da HER2+ a cikin masu ɗaukar PV na BRCA1 da BRCA2 sun yi daidai da bayanan adabi na yanzu.16,43
Sai mu mayar da hankali kan nau'in da wurin BRCA1/2 PV. A cikin ƙungiyarmu, BRCA1 PV da aka fi sani shine c.5035_5039delCTAAT. Kodayake Incorvaia et al. ba su bayyana wannan bambancin a cikin ƙungiyar Sicilian ba, wasu marubuta sun ba da rahotonsa a matsayin ƙwayayen BRCA1 PV.34 An sami PVs da yawa na BRCA1 a cikin ƙungiyarmu - misali c.181T>G, c.514del, c.3253dupA da c.5266dupC - waɗanda aka lura a Sicily.28 Daga cikin waɗannan, ana samun maye gurbi guda biyu na waɗanda suka kafa BRCA1 (c.181T>G da c.5266dupC) a cikin Yahudawan Ashkenazi na Gabashin da Tsakiyar Turai (Poland, Czech), Slovenia, Austrian, Hungary, Belarusian da Jamusanci), 44,45 kuma, a Amurka da Argentina, kwanan nan an ayyana shi a matsayin "bambancin germline mai maimaitawa" a cikin marasa lafiya na Italiya da ke da BC da OC. An riga an gano bambancin 34c.514del a cikin marasa lafiya 8 na ciwon nono daga arewacin Sicily a Palermo da Messina. Abin sha'awa, har ma da Incorvaia et al. An gano bambancin c.3253dupA a wasu iyalai a Catania.28 Mafi yawan wakilan BRCA2 PVs sune c.428dup, c.5851_5854delAGTT da kuma bambancin intronic c.8487+1G>A, waɗanda aka ruwaito dalla-dalla 28 a cikin wani majiyyaci a Palermo tare da c.428dup, c.5851_5854delAGTT PV an lura da shi a cikin gidaje a arewa maso yammacin Sicily, galibi a yankunan Trapani da Palermo, yayin da aka lura da c.5851_5854delAGTT PV a cikin gidaje a arewa maso yammacin Sicily. An fi samun bambancin 8487+1G a cikin mutane daga Messina, Palermo, da Caltanissetta.28 Rebbeck et al. A baya an bayyana canjin c.5851_5854delAGTT a Colombia.37 An sami wani BRCA2 PV, c.631+1G>A, a cikin marasa lafiya na BC da OC daga Sicily (Agrigento, Siracusa da Ragusa).28 Abin lura, mun lura da kasancewar nau'ikan BRCA2 guda biyu (BRCA2 c.631G>A da c.7008-2A>T) tare a cikin wannan majiyyaci, wanda muka ɗauka an raba shi a yanayin cis, kamar yadda aka ruwaito a baya kamar haka.34,46 Waɗannan maye gurbi na BRCA2 galibi ana lura da su a yankin Italiya kuma an gano suna gabatar da codons na dakatarwa da wuri, suna shafar haɗin RNA na manzo kuma suna haifar da gazawar furotin na BRCA2.47,48
Mun kuma zana taswirar BRCA1 da BRCA2 PVs a yankunan OCCR da BCCR na yankunan furotin da kwayoyin halitta. Rebbeck da abokan aikinsa sun bayyana waɗannan yankuna a matsayin wuraren haɗari don kamuwa da cutar kansar mahaifa da nono, bi da bi.49 Duk da haka, shaidar da ke tattare da alaƙar da ke tsakanin wurin da bambance-bambancen ƙwayoyin cuta da haɗarin cutar kansar nono ko ovarian suka ci gaba da zama abin jayayya.28,50-52 A cikin al'ummarmu, BRCA1 PVs sun fi yawa a yankin BCCR, yayin da BRCA2 PVs sun fi yawa a yankin OCCR. Duk da haka, ba mu sami wata alaƙa tsakanin yankunan OCCR da BCCR da siffofin BC ba. Wannan yana iya zama saboda ƙarancin adadin marasa lafiya da ke da maye gurbi na BRCA1/2. Daga mahangar yankin furotin, BRCA1 PVs suna rarraba tare da dukkan furotin, kuma ana samun canje-canjen BRCA2 a yankin maimaita BRC.
A ƙarshe, mun haɗa siffofin ilimin likitancin BC da BRCA1/2 PV. Saboda ƙarancin adadin marasa lafiya da aka haɗa, mun sami babban alaƙa tsakanin Ki-67 da matakin ƙari. Duk da cewa kimantawa da fassarar Ki-67 sun kasance masu rikitarwa, tabbas ne cewa yawan yaduwar yana da alaƙa da ƙaruwar haɗarin sake dawowar cuta da raguwar rayuwa. Zuwa yanzu, matakin da za a iya bambance tsakanin Ki-67 "mai girma" da "ƙananan" shine 20%. Duk da haka, wannan matakin bai shafi yawan marasa lafiya da ke ɗauke da maye gurbi na BRCA1/2 ba, wanda ke da matsakaicin ƙimar Ki-67 na 25%. Wannan yanayin a cikin yawan Ki-67 mai girma za a iya bayyana shi ta hanyar yawan kamuwa da cutar a cikin ƙungiyoyin mu na luminal B da TNBC, waɗanda ƙananan ƙwayoyin A masu haske ke nan. Duk da haka, wasu shaidu suna nuna cewa mafi girman yanke Ki-67 (25-30%) na iya raba marasa lafiya bisa ga hasashensu.53,54 Daga sakamakon bincikenmu, babban alaƙa ba abin mamaki bane. Yana faruwa tsakanin babban Ki-67 da maki da kasancewar BRCA1 PV. A gaskiya ma, ciwon daji masu alaƙa da BRCA1 suna kama da na TNBC kuma suna nuna siffofi masu ƙarfi.16,17
A ƙarshe, wannan binciken ya bayar da rahoto kan matsayin maye gurbin BRCA1/2 a cikin ƙungiyar BC daga gabashin Sicily. Gabaɗaya, bincikenmu ya yi daidai da shaidar da ta riga ta kasance, duka dangane da yawan maye gurbi da kuma siffofin asibiti a cikin BC. Ana buƙatar ƙarin bincike a cikin manyan yawan marasa lafiya na BC masu maye gurbi na BRCA1/2, kamar amfani da nazarin maye gurbi da aka faɗaɗa da yawa, don tantance kasancewar PVs waɗanda suka bambanta kuma ba su da yawa fiye da BRCA1/2. Wannan zai ba da damar gano da kuma kula da yawan mutanen da ke cikin haɗarin kamuwa da cutar kansa saboda maye gurbi na kwayoyin halitta.
Mun tabbatar da cewa marasa lafiya sun sanya hannu kan sanarwar amincewa da sakin samfuran ƙari ba tare da suna ba don dalilai na bincike. Duk marasa lafiya sun sanya hannu kan sanarwar izini a rubuce bisa ga Sanarwar Helsinki. Dangane da manufar AOU Policlinico “G.Rodolico – S.Marco”, an keɓe wannan binciken daga bita na ɗabi'a saboda an gudanar da binciken BRCA1/2 bisa ga aikin asibiti kuma duk marasa lafiya sun ba da izini a rubuce. Marasa lafiya kuma sun yarda da amfani da bayanan su don dalilai na bincike.
Muna godiya ga Farfesa Paolo Vigneri saboda taimakon da ya bayar wajen kula da marasa lafiya da ke fama da cutar kansar mama kamar yadda Kwamitin Da'a ya nema.
Federica Martorana ta bayar da rahoton girmamawa daga Istituto Gentili, Eli Lilly, Novartis, da Pfizer. Sauran marubutan sun bayyana cewa babu wani rikici na sha'awa a cikin wannan aikin.
1. Sung H, Ferlay J, Siegel RL, da sauransu. Kididdigar Ciwon Daji ta Duniya 2020: GLOBOCAN ta kiyasta yawan kamuwa da mace-mace na cutar kansa 36 a ƙasashe 185 a duniya. CA Cancer J Clin.2021;71(3):209-249.doi: 10.3322/caac.21660