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作者 Stella S, Vitale SR, Martorana F, Massimino M, Pavone G, Lanzafame K, Bianca S, Barone C, Gorgone C, Fichera M, Manzella L
UStefania 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 2, 1 Ikliniki yeLizwe yeLizwe, 1 yeMaperimental, iMaperimental 7 iMaperimental, iMaperimental Clinic IYunivesithi yaseCatania, Catania, 95123, Italy;2 Iziko le-2 ye-Experimental Oncology kunye ne-Hematology, i-AOU Policlinico "G.Rodolico - San Marco", Catania, 95123, Italy; I-3 ye-Oncology yezoNyango, i-AOU Policlinico "G. Rodolico - San Marco", Catania, 95123, Italy; 4 iGenetics yezoNyango, ARNAS Garibaldi, Catania, 95123, Italy; 5 Medicine Genetics, ASP, Syracuse, 96100, Italy; 6 ISebe lezeNzululwazi zeBiomedical kunye neBiotechnology, iYunivesithi yaseCatania, iMedical Genetics, eCatania, e-Itali, 95123; 7Oasi Research Institute-IRCCS, eTroina, 94018, e-Itali Unxibelelwano: Stefania Stella, umnxeba +39 095 378 1946, i-imeyile [email protected]; [email protected] Injongo: Utshintsho lwe-germline kwi-BRCA1 kunye ne-BRCA2 kunye nomhlaza webele osekwe (BC), i-ovary (OC) kunye nezinye ezinxulumene nomngcipheko wobomi bonke womhlaza. Uvavanyo lwe-BRCA gene lubalulekile ekuvavanyeni umngcipheko womntu ngamnye, kunye nokufumana iindlela zokuthintela kubantu abanempilo kunye nonyango lokulungisa izigulane zomhlaza. Ukuxhaphaka kotshintsho lwe-BRCA1 kunye ne-BRCA2 kwahluka kakhulu kwiindawo zejografi, kwaye nangona idatha ikho kwiindidi ze-BRCA ezibangela izifo kwiintsapho zaseSicilian, izifundo ezijolise ngokukodwa kubemi basempuma yeSicily azikho. Injongo yophando lwethu yayikukuphanda ukwenzeka kunye nokusasazwa kotshintsho lwe-BRCA olubangela izifo kwi-cohort yezigulane zase-BC ezivela empuma yeSicily kunye nokuvavanya ukudibana kwazo neempawu ezithile ze-BC kusetyenziswa ulandelelwano lwesizukulwana esilandelayo. Ubukho botshintsho obuhambelana nenqanaba le-tumor kunye ne-proliferation index. IZIPHUMO: Lilonke, izigulane ezingama-35 (9%) zazinotshintsho lwe-BRCA olubangela izifo, ezili-17 (49%) kwi-BRCA1 kunye nezili-18 (51%) kwi-BRCA2. Utshintsho lwe-BRCA1 luxhaphakile kwizigulane ze-BC ezine-triple-negative, ngelixa utshintsho lwe-BRCA2 luxhaphake kakhulu kwizigulane ze-luminal BC. Thelekisa kunye nabantu abangenazo iintsholongwane, abantu abaneentsholongwane ze-BRCA1 babenezinga eliphezulu kakhulu le-tumor kunye ne-proliferative index. Izigqibo: Iziphumo zethu zibonelela ngesishwankathelo sesimo sokuguqulwa kwe-BRCA kwizigulane ze-BC ezivela empuma yeSicily kwaye ziqinisekisa indima yohlalutyo lwe-NGS ekuchongeni izigulane ezine-BC yelifa. Ngokubanzi, ezi datha ziyahambelana nobungqina bangaphambili obuxhasa ukuhlolwa kwe-BRCA ukuze kuthintelwe kwaye kunyangwe umhlaza ngokufanelekileyo kubantu abathwala intsholongwane.
Umhlaza webele (BC) lolona hlobo lomhlaza luxhaphakileyo kwihlabathi liphela kwaye lolona hlobo lomhlaza lubulalayo kubasetyhini.1 Iimpawu zebhayoloji ezimisela ukubikezela kwe-BC kunye nokuziphatha kweklinikhi ziye zafundwa kakhulu kwaye zacaciswa kancinci ngokuhamba kwexesha. Enyanisweni, iimpawu ezininzi ezisetyenziselwa ukwahlula i-BC kwiintlobo ezahlukeneyo zeemolekyuli. Ziyi-estrogen (ER) kunye/okanye i-progesterone receptor (PgR), i-human epidermal growth factor receptor 2 (HER2) amplification, i-proliferation index Ki-67 kunye ne-tumor grade (G).2 Ukudibana kwezi zinto ziguquguqukileyo kuchonge ezi ndidi zilandelayo ze-BC: 1) Ii-Luminal tumors, ezibonisa i-ER kunye/okanye i-PgR expression, zibalwa kwi-75% yee-BCs. Ezi tumors zahlulwe ngakumbi zaba yi-Luminal A, xa i-Ki-67 yayingaphantsi kwe-20% kunye ne-HER2 negative, kunye ne-Luminal B, xa i-Ki-67 yayilingana okanye ingaphezulu kwe-20% kwaye xa kukho i-HER2 amplification, nokuba i-proliferation index ingakanani; 2) Iithumba zeHER2+ ezi-ER kunye ne-PgR negative kodwa zibonisa ukwanda kweHER2. Eli qela libandakanya i-10% yazo zonke iithumba zamabele; 3) Umhlaza webele one-triple-negative (TNBC), ongabonisi ukubonakaliswa kwe-ER kunye ne-PgR kunye nokwanda kwe-HER2, ubandakanya malunga ne-15% yomhlaza wamabele. 2-4
Phakathi kwezi ntlobo ze-BC, inqanaba le-tumor kunye ne-proliferation index zimele ii-biomarkers ezinqamlezileyo ezinxulunyaniswa ngokuthe ngqo nangokuzimeleyo nobukhali be-tumor kunye ne-prognosis.5,6
Ukongeza kwezi mpawu zebhayoloji zikhankanyiweyo apha ngasentla, indima yotshintsho lwemfuza olufunyenwe njengelifa olukhokelela ekuphuhlisweni kwe-BC iye yabaluleka ngakumbi kwiminyaka embalwa edlulileyo.7 Malunga ne-1 kwi-10 yeethumba zamabele zizuzwe njengelifa ngenxa yotshintsho lwe-germline kwiijini ezithile.8 Izifundo ezibini ezinkulu ze-epidemiological ezibandakanya abafazi abangaphezu kwe-180,000 kutshanje zichonge iqela leejini ezisibhozo (oko kukuthi, i-ATM, i-BARD1, i-BRCA1, i-BRCA2, i-CHK2, i-PALB2, i-RAD51C, kunye ne-RAD51D) ezinoxanduva lwe-BC yefa. Phakathi kwezi jini, i-BRCA1 kunye ne-BRCA2 (ebizwa ngokuba yi-BRCA1/2) zibonise ulwalamano oluqinileyo nophuhliso lweethumba zamabele.9-12 Enyanisweni, utshintsho lwe-germline BRCA1/2 luyandisa kakhulu umngcipheko wobomi be-BC kunye nezinye izifo ezinobungozi, kubandakanya i-ovarian, i-prostate, i-pancreatic, i-colorectal, kunye ne-melanoma.Ukususela kwiminyaka eli-13 ukuya kwengama-80 ubudala, ukwenzeka kwe-BC yi-72% kubasetyhini abane-BRCA1 pathogenic variant (PV) kunye ne-69% kubasetyhini abane- I-BRCA2 PV.14
Okuphawulekayo kukuba, impapasho yakutshanje ibonisa ukuba umngcipheko we-BC uxhomekeke kuhlobo lwe-PV. Enyanisweni, xa kuthelekiswa notshintsho olubangela ukudumba kwe-pathogenic, utshintsho olucacileyo olungekho ngqiqweni, ingakumbi kwi-gene ye-BRCA1, lunxulunyaniswa nomngcipheko oncitshisiweyo we-BC, ingakumbi kwabasetyhini abadala.15
Ubukho be-BRCA1 okanye i-BRCA2 PV budibene neempawu ezahlukeneyo zebhayoloji kunye nezonyango.16,17 Ii-BC ezinxulumene ne-BRCA1 zihlala zinobundlongondlongo ngokwezonyango, azihlukanga kakuhle, kwaye ziyanda kakhulu. Ezi tumors zihlala zi-triple negative kwaye ziqala kwangethuba. Ii-tumors ezenzeka kwizigulane eziguqukileyo ze-BRCA2 zihlala zibonisa amabakala aphakathi ukuya kwahluka kakuhle kunye ne-variable proliferative indices. Ezi tumors zixhaphake kakhulu kwi-lumen B kwaye zihlala zenzeka kubantu abadala.16-18 Okuphawulekayo kukuba, utshintsho kwi-BRCA1 kunye ne-BRCA2 lunyusa uvakalelo kunyango oluthile, kubandakanya iityuwa zeplatinum kunye namayeza ajoliswe kuwo afana ne-poly(ADP-ribose) polymerase inhibitors (PARPi).19,20
Kwiminyaka embalwa edlulileyo, ukuphunyezwa kwe-next-generation sequencing (NGS) kwiklinikhi kuye kwenza ukuba inani elikhulayo lezigulane ze-BC livavanywe ngokwemolekyuli ukuze lifumane iimpawu zomhlaza, kuquka i-BRCA1/2.21 Kwangaxeshanye, iinkcazo ezisekelwe kwiikhrayitheriya ezichanekileyo malunga nembali yosapho, idemografi, kunye neempawu zeklinikhi ukuze kuchongwe ngcono abantu abafanelekileyo kuvavanyo lwe-BRCA1/2.22,23 Kulo mongo, ubungqina buqokelelana kuvavanyo lwe-BRCA1/2 kumaqela athile, bubonisa umahluko kwiindawo ezahlukeneyo zejografi.24–27 Nangona kukho iingxelo malunga neqela le-BC entshona yeSicily, zimbalwa iinkcukacha ezikhoyo kuvavanyo lwe-BRCA1/2 kubemi basempuma yeSicily.28,29
Apha sichaza iziphumo zovavanyo lwe-germline BRCA1/2 kwizigulane zase-BC ezivela empuma yeSicily, nto leyo edibanisa ngakumbi ubukho be-BRCA1 okanye i-BRCA2 mutations kunye neempawu eziphambili zeklinikhi zezi thumba.
Uphononongo olujonga emva lwenziwe kwi-"Center for Experimental Oncology and Hematology" kwiSibhedlele iPoliclinico.Rodolico – San Marco eCatania. Ukusukela ngoJanuwari 2017 ukuya kuMatshi 2021, izigulane ezingama-455 ezinesifo somhlaza wamabele kunye ne-ovarian, i-melanoma, i-pancreatic okanye i-prostate zathunyelwa kwilebhu yethu yokuxilongwa kwe-molecular ukuze zivavanywe ngokwemfuza ye-BRCA/2. Olu phononongo lwenziwe ngokuhambelana neSibhengezo saseHelsinki, kwaye bonke abathathi-nxaxheba banike imvume ebhaliweyo ngaphambi kohlalutyo lwe-molecular.
Iimpawu ze-histological nezebhayoloji (ER, PgR, imeko ye-HER2, Ki-67, kunye nodidi) ze-BC zihlolwe kwi-biopsy eyintloko okanye kwiisampuli zotyando, kuthathelwa ingqalelo kuphela izinto ezinoburhalarhume obunamandla. Ngokusekelwe kwezi mpawu, ii-BC zahlulwe ngolu hlobo lulandelayo: i-luminal A (ER+ kunye/okanye i-PgR+, HER2-, Ki-67<20%), i-luminal B (ER+ kunye/okanye i-PgR+, HER2-, Ki-67≥20%), i-luminal B-HER2+ (ER kunye/okanye i-PgR+, HER2+), i-HER2+ (ER kunye ne-PgR-, HER2+) okanye i-triple negative (ER kunye ne-PgR-, HER2-).
Ngaphambi kokuvavanya imeko yokuguquka kwe-BRCA1 kunye ne-BRCA2, iqela elinamacandelo amaninzi kuquka ingcali ye-oncologist, ingcali ye-genetic, kunye nengcali yezengqondo benze ingcebiso ye-tumor genetics kwisigulane ngasinye ukuze kuqinisekiswe ukuba kukho i-BRCA1 kunye/okanye i-BRCA1. okanye abantu abanomngcipheko ophezulu we-PV kwi-BRCA2 gene. Ukukhethwa kwesigulane kwenziwe ngokwezikhokelo ze-Italian Society of Medical Oncology (AIOM) kunye neengcebiso zaseSicilian zasekuhlaleni.30,31 Ezi khrayitheriya ziquka: (i) imbali yosapho yeentlobo ezahlukeneyo ze-pathogenic ezaziwayo kwiijini ezinokubangela ukucaphuka (umz., i-BRCA1, i-BRCA2, i-TP53, i-PTEN); (ii) amadoda ane-BC; (iii) lawo ane-BC kunye ne-OC; (iv) abafazi abane-BC <36 iminyaka, i-TNBC <60 iminyaka, okanye i-BC <50 iminyaka; (v) imbali yezonyango zomntu ye-BC <50 iminyaka kunye nosapho olunye ubuncinane lwenqanaba lokuqala: (a) i-BC <50 iminyaka; (b) i-OC engekho mucinous kwaye engekho borderline yalo naliphi na ubudala; (c) i-BC yamacala omabini; (d) i-BC yamadoda; (e) umhlaza wepancreatic; (f) umhlaza weprostate; (vi) imbali yomntu ye-BC engaphezulu kweminyaka engama-50 kunye nembali yosapho lwe-BC, i-OC, okanye umhlaza wepancreatic kwizalamane ezizalamane ezinezinga lokuqala omnye komnye (kuquka izalamane ezizalamane nazo ezinezinga lokuqala); (vii) Imbali yomntu ye-OC kunye nesalamane esinye ubuncinane sezinga lokuqala: (a) BC <50 iminyaka; (b) NOC; (c) i-BC yamacala omabini; (d) indoda ye-BC; (vii) ibhinqa eline-OC yezinga eliphezulu.
Isampuli yegazi eliyi-20 mL elivela kumguli ngamnye laza laqokelelwa kwiityhubhu ze-EDTA (BD Biosciences). I-Genomic DNA yahlulwa kwiisampuli zegazi elipheleleyo eziyi-0.7 mL kusetyenziswa i-QIAsymphony DSP DNA Midi kit Isolation Kit (QIAGEN, Hilden, Italy) ngokwemiyalelo yomenzi yaza yadlula kwi-Qubit® 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Yenza umlinganiselo. Ukuphucula iithagethi kunye nokulungiselela ithala leencwadi kwenziwa yi-Oncomine™ BRCA Research Assay Chef, elungele ukulayishwa kwi-Ion AmpliSeq™ Chef Reagents DL8 Kit yokulungiselela ithala leencwadi ngokuzenzekelayo ngokwemiyalelo yomenzi. Le khithi inee-multiplex PCR primer pools ezimbini ezinokusetyenziselwa ukufunda zonke ii-genes ze-BRCA1 (NM_007300.3) kunye ne-BRCA2 (NM_000059.3). Ngamafutshane, i-15 µL yesampulu nganye ye-DNA (10 ng) yongezwe kwiiplate ezineebhakhowudi zokulungiselela ithala leencwadi kwaye zonke ii-reagents kunye nezinto ezisetyenziswayo zalayishwa kwisixhobo se-Ion Chef™. Ukulungiswa kwethala leencwadi ngokuzenzekelayo kunye nokudibanisa isampuli yethala leencwadi elineebhakhowudi kwenziwa kwisixhobo se-Ion Chef™. Inani leelayibrari ezilungiselelweyo lavavanywa yi-Qubit® 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA) ngokwemiyalelo yomenzi. Okokugqibela, iilayibrari zidityaniswe kwimilinganiselo elinganayo kwiityhubhu zesampulu zethala leencwadi le-Ion Chef™. (iityhubhu ezineebhakhowudi) kwaye zifakwe kwisixhobo se-Ion Chef™. Ulandelelwano lwenziwe kusetyenziswa isixhobo se-Ion Torrent S5 (Thermo Fisher Scientific) (Thermo Fisher Scientific) kusetyenziswa i-Ion 510 Chip (Thermo Fisher Scientific). Uhlalutyo lwedatha lwenziwe yi-Amplicon Suite (SmartSeq srl) kunye ne-Ion Reporter Software.
Zonke iintlobo zokwahlulwa kwamagama zilandele izikhokelo zangoku zeHuman Genome Variation Consortium, efumaneka kwi-intanethi (HGVS, http://www.hgvs.org/mutnomen). Ukubaluleka kweklinikhi kweentlobo zokwahlulwa kweBRCA1/2 kuchazwe kusetyenziswa ukuhlelwa kweInternational Consortium ENIGMA (Evidence-Based Network for Interpreting Germline Mutant Alleles, https://enigmaconsortium.org/) kunye nokubonisana needathabheyisi ezahlukeneyo ezifana ne-ARUP, BRCAEXCHANGE, ClinVar, IARC_LOVD, kunye ne-UMD. Ukuhlelwa kuquka iindidi ezintlanu ezahlukeneyo zomngcipheko: ezingalunganga (udidi I), ezinokwenzeka ukuba azilunganga (udidi II), ezingafaniyo nokuba zibalulekile (VUS, udidi III), ezinokwenzeka ukuba zibangela izifo (udidi IV), kunye nezifo (udidi V). Abanye bahlalutye nefuthe lokuguqulwa kwezakhi zofuzo kwisakhiwo kunye nomsebenzi weprotheni, isixhobo esinolwazi esinokufikelela kwiidathabheyisi ezingama-30.32
Ukunika i-VUS nganye ukubaluleka kweklinikhi okunokwenzeka, kusetyenziswe ezi algorithms zilandelayo zokuqikelela iiproteni zokubala: I-MUTATION TASTER, 33 PROVEAN-SIFT (http://provean.jcvi.org/index.php), i-POLYPHEN-2 (http:// /genetics.bwh.harvard.edu/pph2/) kunye ne-Align-GVGD (http://agvgd.hci.utah.edu/agvgd_input.php). Iindidi ezihlelwe njengeklasi yoku-1 neyesi-2 zithathwa njengeentlobo zasendle.
Ukulandelelana kweSanger kuqinisekisile ukubakho kohlobo ngalunye lwe-pathogenic. Ngamafutshane, isibini see-primers ezithile zenzelwe uhlobo ngalunye olufunyenweyo ngokusebenzisa ulandelelwano lwe-BRCA1 kunye ne-BRCA2 gene reference (NG_005905.2, NM_007294.3 kunye ne-NG_012772.3, NM_000059.3, ngokulandelelana). Ke ngoko, i-PCR ejoliswe kuyo yenziwe yalandelwa lulandelelwano lweSanger.
Izigulane ezivavanywe zingenayo i-gene ye-BRCA1/2 zivavanywe nge-multiplex ligation-dependent probe amplification (MLPA) ngokwemiyalelo yomenzi ukuvavanya ubukho be-large genomic rearrangements (LGR). Ngamafutshane, iisampulu ze-DNA ziyasuswa kwaye kusetyenziswa ii-probes ze-gene ezifikelela kuma-60 ze-BRCA1 kunye ne-BRCA2, nganye ibona ulandelelwano oluthile lwe-DNA malunga ne-nucleotides ezingama-60 ubude. Iimveliso ze-probe amplification, eziquka iseti eyahlukileyo ye-PCR amplicons, emva koko zahlalutywa yi-capillary electrophoresis kunye ne-Cofalyser.Net software kunye neetafile ze-Cofalyser ezifanelekileyo ze-batch-specific (www.mrcholland.com).
Iinguqu ezikhethiweyo zeklinikhi (ibanga le-histological kunye ne-Ki-67% proliferation index) zidibene nobukho be-BRCA1/2 PV, zibalwe kusetyenziswa isoftware yePrism v. 8.4 kusetyenziswa uvavanyo oluchanekileyo lukaFisher oluthatha ixabiso le-p <0.05 libalulekile.
Phakathi kukaJanuwari 2017 noMatshi 2021, izigulana ezingama-455 zahlolwa i-germline BRCA1/2 mutations. Uvavanyo lokuguqulwa kwezakhi zofuzo lwenziwe kwiZiko leSibhedlele iPoliclinico's Center for Experimental Oncology and Hematology. Ngokwesikhokelo saseSicilian (http://www.gurs.regione.sicilia.it/Indicep1.htm, N. 02-Venerdì 10 Gennaio 2020), iRodolico yaseCatania – San Marco iyonke, izigulana ezingama-389. Kwakukho umhlaza webele, umhlaza wesibeleko ongama-37, umhlaza wepancreatic ongama-16, umhlaza weprostate osi-8 kunye ne-melanoma ezi-5. Ukusasazwa kwezigulana ngokohlobo lomhlaza kunye neziphumo zohlalutyo kuboniswe kuMfanekiso 1.
Umfanekiso 1 ubonisa itshathi yendlela ebonisa isishwankathelo sophando. Izigulane ezineethumba zebele, i-melanoma, i-pancreatic, i-prostate, okanye i-ovarian zivavanywe utshintsho kwi-BRCA1 kunye ne-BRCA2 genes.
Izifinyezo: ii-PV, uhlobo olubangela izifo; i-VUS, uhlobo olungaqinisekanga; i-WT, ulandelelwano lwe-BRCA1/2 yohlobo lwasendle.
Sigxile ngokukhetha kwizifundo zethu kwiiqela zomhlaza webele. Izigulane zazineminyaka engama-49 ubudala (uluhlu lwama-23-89) kwaye uninzi lwazo yayingabasetyhini (n=376, okanye ama-97%).
Kwaba bantu, abangama-64 (17%) babene-BRCA1/2 mutations kwaye bonke babengabasetyhini. Amashumi amathathu anesihlanu (9%) babene-PV kwaye abangama-29 (7.5%) babene-VUS. Ishumi elinesixhenxe (48.6%) kwiindidi ezingama-35 ze-pathogenic zenzeka kwi-BRCA1 kunye ne-18 (51.4%) kwi-BRCA2, ngelixa ama-5 e-VUS enzeka kwi-BRCA1 (17.2%) kunye nama-24 (82.8%) kwi-BRCA2 (Imifanekiso 1 kunye no-2). I-LGR yayingekho kuhlalutyo lwe-MLPA.
Umfanekiso 2. Uhlalutyo lwe-BRCA1 kunye ne-BRCA2 mutations kwizigulane ezingama-389 zomhlaza webele.(A) Ukusasazwa kweentlobo ezahlukeneyo ze-pathogenic (PV) (ezibomvu), iintlobo ezahlukeneyo ezingaqinisekanga (VUS) (orenji), kunye ne-WT (eluhlaza okwesibhakabhaka) kwizigulane ezingama-389 zomhlaza webele; (B) Izigulane ezingama-389 zomhlaza webele Amashumi amathathu anesihlanu (9%) ayeneentlobo ezahlukeneyo ze-BRCA1/2 ze-pathogenic (ii-PV).Phakathi kwabo, abali-17 (48.6%) babene-BRCA1 PV carriers (ebomvu obumnyama) kwaye abali-18 (51.4%) babene-BRCA2 carriers (ebomvu okhanyayo); (C) Abantu abangama-29 (7.5%) kwabangama-389 ababene-VUS, ii-genes ezi-5 (17.2%) ze-BRCA1 (orenji obumnyama) kunye nama-24 (82.8%) ze-BRCA2 genes (orenji okhanyayo).
Izifinyezo: ii-PV, uhlobo olubangela izifo; i-VUS, uhlobo olungaqinisekanga; i-WT, ulandelelwano lwe-BRCA1/2 yohlobo lwasendle.
Emva koko sihlolisise ukuxhaphaka kwe-BC molecular subtypes kwizigulane ezine-BRCA1/2 PV. Ukusasazwa kwaquka i-2 (5.7%) luminal A, i-15 (42.9%) luminal B, i-3 (8.6%) luminal B-HER2+, i-2 (5.7%) HER2+ kunye ne-13 (37.1%) yezigulane ze-TNBC. Phakathi kwezigulane ezine-BRCA1, i-5 (29.4%) yayine-luminal B BC, i-2 (11.8%) yayinesifo se-HER2+, kwaye i-10 (58.8%) yayine-TNBC. Ii-tumor ezingenazo iinguqu ze-BRCA1 yayizi-luminal A okanye i-luminal B-HER2+ (Umfanekiso 3). Kwiqela elincinci le-BRCA2-positive, ii-tumor ezili-10 (55.6%) zazizi-luminal B, i-3 (16.7%) yayizi-luminal B-HER2+, i-3 (16.7%) TNBC kunye ne-2 (11.1%) yayizi-luminal A (Umfanekiso 3). Hayi. Iithumba zeHER2+ bezikho kweli qela. Ngoko ke, utshintsho lweBRCA1 luxhaphakile kwizigulane zeTNBC, ngelixa utshintsho lweBRCA2 luxhaphakile kubantu abane-lumen B.
Umfanekiso 3 Ukuxhaphaka kweentlobo ezahlukeneyo zomhlaza webele kwizigulane ezineentlobo ezahlukeneyo ze-BRCA1 kunye ne-BRCA2. IiHistogram ezibonisa ukusasazeka kwe-BRCA1- (ubomvu obumnyama) kunye ne-BRCA2- (ubomvu obukhanyayo) ii-PV phakathi kweentlobo ezahlukeneyo zeemolekyuli zezigulane zomhlaza webele. Amanani abikiweyo kwibhokisi nganye amele ipesenti yezigulane ezine-BRCA1 kunye ne-BRCA2 PV kuhlobo ngalunye lomhlaza webele.
Izifinyezo: ii-PV, uhlobo lwe-pathogenic; i-HER2+, i-human epidermal growth factor receptor 2 positive; i-TNBC, umhlaza webele one-triple-negative.
Emva koko, sivavanye uhlobo kunye nendawo yezakhi zofuzo ze-BRCA1 kunye ne-BRCA2 PVs. Kwi-BRCA1 PV, sibone ii-single nucleotide variants ezi-7 (ii-SNV), ukususwa oku-6, ukuphinda-phinda oku-3 kunye nokufakwa oku-1. Inguqu enye kuphela (c.5522delG) imele ukufunyanwa okutsha. Eyona PV iqhelekileyo ye-BRCA1 efunyenweyo kuzo zombini izifundo yayiyi-c.5035_5039delCTAAT. Olu tshintsho lubandakanya ukususwa kwee-nucleotides ezintlanu (CTAAT) kwi-BRCA1 exon 15, nto leyo ebangele ukuba i-amino acid leucine ithathelwe indawo yi-tyrosine kwi-codon 1679, kwaye ngenxa yokutshintsha kwe-frameshift ene-alternative stop codon eqikelelweyo ikhokelela ekunqunyulweni kweproteni ngaphambi kwexesha. Zonke ezinye iinguqu zifunyanwa kwimeko enye kuphela. Okuphawulekayo kukuba, enye yee-PV ezixeliweyo yayikwindawo yesivumelwano se-splice site (c.4357+1G>T) (Itheyibhile 1).
Ngokuphathelele i-BRCA2 PV, sibone ukucinywa oku-6, ii-SNV ezi-6 kunye nokuphindaphindwa oku-2. Akukho nanye kwezi nguqu zifunyenweyo intsha. Iinguqu ezintathu ziphinde zavela kuluntu lwethu, i-c.428dup kunye ne-c.8487+1G>A ezibonwe kubantu aba-3, zilandelwa yi-c.5851_5854delAGTT ezifunyenwe kwiimeko ezimbini. Utshintsho lwe-c.428dup lubandakanya ukuphindaphindwa kwe-C kwi-exon 5 ye-BRCA2, eqikelelwa ukuba iza kufaka iproteni enqunyulweyo, engasebenziyo. Utshintsho lwe-c.8487+1G>A lwenzeka kummandla we-intronic we-BRCA2 intron 19 (± 1,2) kwaye luchaphazela ulandelelwano lwe-splicing consensus, nto leyo ebangela ukutshintsha kwe-splicing okubangela iproteni engaqhelekanga okanye engekhoyo. Uhlobo lwe-c.5851_5854delAGTT pathogenic lubangelwa kukususwa kwe-nucleotide ye-4 kwiindawo ze-nucleotide 5851 ukuya kwi-5854 kwi-exon 10 yekhowudi I-gene ye-BRCA2 kwaye ibangela utshintsho lwefreyimu oluguqulelweyo nge-alternative stop codon eqikelelweyo (p.S1951WfsTer). Okuphawulekayo kukuba, njengoko bekuxeliwe ngaphambili, zombini ezi nguqu c.631G>A kunye ne-c.7008-2A>T zifunyenwe kwisigulana esinye.34 Utshintsho lokuqala lubandakanya ukutshintshwa kwe-adenosine (A) kwi-exon 7 ye-BRCA2 nge-guanine (G) equlethe i-nucleotide okubangela utshintsho lwe-valine kwi-isoleucine kwi-codon 211, i-isoleucine I-Amino acid yi-amino acid eneempawu ezifanayo kakhulu. Olu tshintsho luchaphazela i-mRNA splicing eqhelekileyo. Uhlobo lwesibini lukwindawo ye-intronic kwaye lubangela ukutshintshwa kwe-A kabini kwi-thymine (T) ngaphambi kwe-exon 13 ye-gene encoding BRCA2. Utshintsho lwe-c.7008-2A>T lunokuvelisa ii-transcripts ezininzi zobude obahlukeneyo. Ngaphezu koko, kwiqela le-BRCA2 PVs, iinguqu ezi-4 kwezili-18 (22.2%) zazi-intronic.
Emva koko simaphe utshintsho olubi lwe-BRCA1/2 kwiindawo ezisebenzayo kunye neendawo ezibopha iiproteni (Umzobo 4). Kwi-gene ye-BRCA1, ama-50% ee-PV afumaneka kummandla weqela lomhlaza webele (i-BCCR), ngelixa ama-22% eenguqu ayefumaneka kummandla weqela lomhlaza we-ovarian (i-OCCR) (Umzobo 4A). Kwi-PV ye-BRCA2, ama-35.7% eenguqu ayefumaneka kummandla we-BCCR kwaye ama-42.8% eenguqu ayefumaneka kwi-OCCR (Umzobo 4B). Okulandelayo, sivavanye indawo ye-PV ngaphakathi kweendawo zeproteni ye-BRCA1 kunye ne-BRCA2. Kwiproteni ye-BRCA1, sifumene ii-PV ezintathu kwiindawo ze-loop kunye ne-coiled coil, kunye ne-mutations ezimbini kwindawo ye-BRCT (Umzobo 4A). Kwiproteni ye-BRCA2, ii-PV ezi-4 zimaphe kwi-domain ye-BRC repeat, ngelixa utshintsho olu-3 lwe-intronic kunye no-3 lwe-exonic lufunyenwe kwiindawo ze-oligo/oligosaccharide-binding (OB) kunye ne-tower (T) (Umzobo 4B).
Umfanekiso 4 Ukubonakaliswa kwesicwangciso seeproteni ze-BRCA1 kunye ne-BRCA2 kunye nokuhlala kweentlobo ezahlukeneyo ze-pathogenic. Lo mfanekiso ubonisa ukusasazeka kweentlobo ezahlukeneyo ze-BRCA1 (A) kunye ne-BRCA2 (B) kwizigulane ezinomhlaza webele. Utshintsho lwe-exonic luboniswe ngombala oluhlaza okwesibhakabhaka, ngelixa iintlobo ze-intronic ziboniswe ngombala o-orenji. Ukuphakama kwebha kubonisa inani lamatyala. Iiproteni ze-BRCA1 kunye ne-BRCA2 kunye neendawo zazo zokusebenza zixeliwe. (A) Iproteni ye-BRCA1 iqulethe i-loop domain (RING) kunye nolandelelwano lwe-nuclear localization (NLS), i-coiled-coil domain, i-SQ/TQ cluster domain (SCD), kunye ne-BRCA1 C-terminal domain (BRCT). (B) Iproteni ye-BRCA2 iqulethe ukuphindaphinda kwe-BRC okusibhozo, i-DNA-binding domain ene-helical domain (Helical), ii-oligonucleotide/oligosaccharide-binding (OB) folds ezintathu, i-tower domain (T), kunye ne-An NLS kwicala le-C. Iindawo ezibizwa ngokuba yi-Breast Cancer Cluster Region (BCCR) kunye ne-Ovarian Cancer Cluster Region (OCCR) ziboniswe kwi ezantsi.*Imela utshintsho olumisela ii-stop codons.
Emva koko siphande iimpawu ze-BC clinicopathological ezinokunxulumana nokuba khona kwe-BRCA1/2 PV. Iirekhodi zeklinikhi ezipheleleyo bezifumaneka kwizigulane ezili-181 ezine-BRCA1/2 (ezingenazo ii-carriers) kunye nabo bonke abathwali (n = 35). Kwakukho unxibelelwano phakathi kwesantya sokukhula kwe-tumor kunye nomgangatho.
Sibale ukusasazwa kweKi-67 ngokusekelwe kumbindi weqela lethu (25%, uluhlu <10-90%). Izifundo ezineKi-67 <25% zichazwe njenge "Ki-67 esezantsi", ngelixa abantu abanexabiso eliyi-≥ 25% bathathwa njenge "Ki-67 ephezulu". Umahluko omkhulu weKi-67 (p<0.01) ufunyenwe phakathi kwabathwali abangengabo abathwali kunye nabathwali be-BRCA1 PV (Umzobo 5A).
Umfanekiso 5 Ulwalamano lweKi-67 kunye nokusasazwa kwamanqanaba kubafazi abanomhlaza webele abane-BRCA1 kunye ne-BRCA2 PVs nabangenayo.(A) Ibhokisi ebonisa amaxabiso aphakathi eKi-67 kwizigulana ze-BC ezingathwaliyo ezili-181 xa kuthelekiswa nezigulana ze-BRCA1 (18) okanye ze-BRCA2 (17) ze-PV. Amaxabiso e-P angaphantsi kwe-0.5 athathwa njengabalulekileyo ngokwezibalo.(B) I-Histogram emele ukwabiwa kwezigulana zomhlaza we-BC kumaqela ebanga le-histological (G2 kunye ne-G3) ngokwesimo sokuguquka kwe-BRCA1 kunye ne-BRCA2 (izifundo ze-WT, izithwali ze-BRCA1 kunye ne-BRCA2 PVs).
Ngokufanayo, sihlolisise ukuba ingaba inqanaba le-tumor lihambelana na nokuba khona kwe-BRCA1/2 PV. Ekubeni i-G1 BC yayingekho kuluntu lwethu, sahlulahlula izigulane zaba ngamaqela amabini (i-G2 okanye i-G3). Ngokuhambelana neziphumo ze-Ki-67, uhlalutyo lutyhile ulwalamano olubalulekileyo phakathi kwenqanaba le-tumor kunye noguquko lwe-BRCA1, kunye nenani eliphezulu lee-tumor ze-G3 kubathwali be-BRCA1 xa kuthelekiswa nabangengabo abathwali (p<0.005) (Umfanekiso 5B).
Inkqubela phambili kwitekhnoloji yokulandelelanisa i-DNA ivumele inkqubela engazange ibonwe ngaphambili kuvavanyo lwe-BRCA1/2 yemfuza, nto leyo enefuthe elibalulekileyo kwizigulane ezinembali yomhlaza kusapho. Ukuza kuthi ga ngoku, malunga neendidi ezingama-20,000 ze-BRCA1/2 zichongiwe kwaye zahlulwa ngokwe-American Society of Medical Genetics 35 kunye nenkqubo ye-ENIGMA.35,36 Kuyaziwa ukuba i-BRCA1/2 mutational spectrum iyahluka kakhulu kwiindawo zejografi.37 Ngaphakathi e-Itali, izinga le-BRCA1/2 PVs lisusela kwi-8% ukuya kwi-37%, libonisa ukuguquguquka okukhulu kwangaphakathi kwelizwe.38,39 Ngabemi abaphantse babe zizigidi ezi-5, iSicily yindawo yesihlanu ngobukhulu e-Itali ngokwenani labemi. Nangona idatha ikho malunga nokusasazwa kwe-BRCA1/2 entshona yeSicily, akukho bungqina buninzi kwinxalenye esempuma yesiqithi.
Uphononongo lwethu lolunye lweengxelo zokuqala malunga nokwanda kwe-BRCA1/2 PV kwizigulane zase-BC empuma yeSicily.28 Sigxile kuhlalutyo lwethu kwi-BC, njengoko esi sesona sifo sixhaphakileyo kwiqela lethu.
Xa bevavanya izigulane ezingama-389 BC, i-9% yayinee-PV ze-BRCA1/2, ezazisasazwa ngokulinganayo phakathi kwe-BRCA1 kunye ne-BRCA2. Ezi ziphumo zihambelana nezo zixelwe ngaphambili kubemi base-Itali.28 Okubangela umdla kukuba, i-3% (13/389) yeqela lethu yayingamadoda. Eli zinga liphezulu kunokuba bekulindelwe kumhlaza webele wamadoda (1% yazo zonke ii-BC),40 zibonisa ukhetho lwethu lwamaqela ngokusekelwe kumngcipheko wokuguquka kwe-BRCA1/2. Nangona kunjalo, akukho namnye kula madoda oye wenza i-BRCA1/2 PV, ngoko ke babengabaviwa bohlalutyo olongezelelweyo lweemolekyuli ukuze kuthintelwe ubukho bezinguquko ezingaqhelekanga ezifana ne-PALB2, RAD51C kunye ne-D, phakathi kwezinye. Iindidi ezingabalulekanga zifunyenwe kwi-7% yabantu apho i-BRCA2 VUS yayibonakala khona. Kwanale miphumo ihambelana nobungqina obukhoyo.28,41,42
Xa sasihlalutya ukusasazwa kwe-BC molecular subtypes kubafazi abaguqukileyo be-BRCA1/2, siqinisekisile unxulumano olwaziwayo phakathi kwe-TNBC kunye ne-BRCA1 PV (58.8%) kunye naphakathi kwe-luminal B BC kunye ne-BRCA2 PV (55.6%).16,43 Ii-luminal A kunye ne-HER2+ tumors kwi-BRCA1 kunye ne-BRCA2 PV carriers ziyahambelana nedatha yoncwadi ekhoyo.16,43
Emva koko sigxila kuhlobo kunye nendawo ye-BRCA1/2 PV. Kwiqela lethu, eyona BRCA1 PV ixhaphakileyo yayiyi-c.5035_5039delCTAAT. Nangona u-Incorvaia et al. Abazange bayichaze le nguqulelo kwiqela labo laseSicilian, abanye ababhali bayixele njenge-germline BRCA1 PV.34 Kufunyenwe ii-PV ezininzi ze-BRCA1 kwiqela lethu – umz. c.181T>G, c.514del, c.3253dupA kunye ne-c.5266dupC – eziye zabonwa eSicily.28 Kwezi, iinguqulelo ezimbini zabasunguli be-BRCA1 (c.181T>G kunye ne-c.5266dupC) zifumaneka rhoqo kumaJuda ase-Ashkenazi aseMpuma naseMbindini Yurophu (ePoland, eCzech), eSlovenia, e-Austria, eHungary, eBelarusian naseJamani), 44,45 kwaye, e-United States naseArgentina, kutshanje ichazwe njenge-"recurrent germline variant" kwizigulana zase-Itali ezine-BC kunye ne-OC. Uhlobo lwe-34c.514del lwaluchongiwe ngaphambili kwizigulana ezi-8 zomhlaza webele ezivela kumantla eSicily ePalermo naseMessina. Okubangel 'umdla kukuba, nkqu no-Incorvaia et al. ifumene uhlobo lwe-c.3253dupA kwezinye iintsapho eCatania.28 Ii-PV ze-BRCA2 ezimele kakhulu yi-c.428dup, c.5851_5854delAGTT kunye nohlobo lwe-intronic c.8487+1G>A, ezixelwe ngokweenkcukacha 28 kwisigulana ePalermo esine-c.428dup, c.5851_5854delAGTT PV ibonwe kwiintsapho ezikumntla-ntshona weSicily, ikakhulu kwimimandla yaseTrapani nasePalermo, ngelixa i-c.5851_5854delAGTT PV ibonwe kwiintsapho ezikumntla-ntshona weSicily. Uhlobo lwe-8487+1G>A luqheleke kakhulu kubantu abavela eMessina, ePalermo, naseCaltanissetta.28 URebbeck et al. ngaphambili ichaze utshintsho lwe-c.5851_5854delAGTT eColombia.37 Enye i-BRCA2 PV, c.631+1G>A, ifunyenwe kwizigulane ze-BC kunye ne-OC ezivela eSicily (Agrigento, Siracusa kunye neRagusa).28 Okuphawulekayo kukuba, sibonile ukubakho kweendidi ezimbini ze-BRCA2 (BRCA2 c.631G>A kunye c.7008-2A>T) kwisigulana esinye, esasicinga ukuba sahlulwe kwimo ye-cis, njengoko bekuxelwe ngaphambili ngolo hlobo.34,46 Ezi nguqu ze-BRCA2 ziqhele ukubonwa kummandla wase-Itali kwaye zifunyenwe ukuba zingenisa ii-codons zokuma ngaphambi kwexesha, zichaphazela i-messenger RNA splicing kwaye zibangela ukuba iproteni ye-BRCA2 ingaphumeleli.47,48
Sikwadwelise ii-PV ze-BRCA1 kunye ne-BRCA2 kwiindawo ze-OCCR kunye ne-BCCR ezibonisa ukuba kukho iiproteni kunye neejini. Ezi ndawo zichazwe nguRebbeck et al. njengeendawo ezinobungozi bokukhula komhlaza we-ovarian kunye nebele, ngokwahlukeneyo.49 Nangona kunjalo, ubungqina malunga nobudlelwane phakathi kwendawo yeentlobo ze-germline kunye nomngcipheko womhlaza webele okanye we-ovarian buhlala buphikisana.28,50-52 Kubemi bethu, ii-PV ze-BRCA1 zazikwindawo ye-BCCR, ngelixa ii-PV ze-BRCA2 zazikwindawo ye-OCCR. Nangona kunjalo, asikwazanga ukufumana naluphi na unxulumano phakathi kweendawo ze-OCCR kunye ne-BCCR ezibonisa ukuba kukho iinguqu kwi-BC kunye neempawu ze-BC. Oku kusenokuba kungenxa yenani elincinci lezigulana ezine-mutations ye-BRCA1/2. Ukusuka kwimbono ye-protein domain, ii-PV ze-BRCA1 zisasazwa kuyo yonke iproteni, kwaye utshintsho lwe-BRCA2 lufumaneka ngokukhethekileyo kwindawo ye-BRC repeat.
Okokugqibela, sidibanise iimpawu ze-BC clinicopathological ne-BRCA1/2 PV. Ngenxa yenani elincinci lezigulana ezibandakanyiweyo, sifumene ulwalamano olubalulekileyo phakathi kwe-Ki-67 kunye nomgangatho wesimila. Nangona uvavanyo kunye nokutolikwa kwe-Ki-67 kusaphikisana kancinci, kuyacaca ukuba amazinga aphezulu okwanda anxulunyaniswa nomngcipheko okhulayo wokuphinda isifo sibuye kunye nokuncipha kokusinda. Ukuza kuthi ga ngoku, umda wokwahlula phakathi kwe-Ki-67 "ephezulu" kunye "nephantsi" yi-20%. Nangona kunjalo, lo mda awusebenzi kwinani lezigulane zethu zokuguqulwa kwe-BRCA1/2, ezinexabiso eliphakathi le-Ki-67 le-25%. Olu tyekelo lwamazinga aphezulu e-Ki-67 lunokuchazwa kukuxhaphaka kwamaqela ethu e-luminal B kunye ne-TNBC, apho kwakukho iithumba ezimbalwa ze-luminal A. Nangona kunjalo, ubungqina obuthile bubonakala bubonisa ukuba umda ophezulu we-Ki-67 (25-30%) unokwahlula ngcono izigulana ngokwesimo sazo.53,54 Kwiziphumo zohlalutyo lwethu, ulwalamano olubalulekileyo alumangalisi. Lwenzeka phakathi kwe-Ki-67 ephezulu kunye namanqanaba kunye nobukho be-BRCA1 I-PV. Enyanisweni, iithumba ezinxulumene ne-BRCA1 ziqhelekile kwi-TNBC kwaye zibonisa iimpawu ezibukhali ngakumbi.16,17
Ukuqukumbela, olu phononongo lubonelela ngengxelo malunga nesimo sokuguquka kwe-BRCA1/2 kwiqela le-BC elivela empuma yeSicily. Ngokubanzi, iziphumo zethu ziyahambelana nobungqina obukhoyo, kokubini ngokwezinga lokwanda kwe-mutation kunye neempawu zeklinikhi kwi-BC. Izifundo ezingaphezulu kumaqela amakhulu ezigulane ze-BRCA1/2-mutant BC, njengokusebenzisa uhlalutyo lwe-multigenome expanded mutational, ziyafuneka ukuvavanya ubukho be-PVs ezahlukileyo kwaye ezingaqhelekanga kune-BRCA1/2. Oku kuya kuvumela ukuchongwa kunye nolawulo olufanelekileyo lwenani elikhulayo labantu abasengozini enkulu yomhlaza ngenxa ye-genetic mutations.
Siqinisekisile ukuba izigulana zisayine imvume yokukhupha iisampulu zazo zethumba ngokungaziwa ngeenjongo zophando. Zonke izigulana zisayine imvume ebhaliweyo ngokweSibhengezo saseHelsinki. Ngokwemigaqo-nkqubo ye-AOU Policlinico “G.Rodolico – S.Marco”, olu phononongo aluzange lujongwe kuphononongo lokuziphatha kuba uhlalutyo lwe-BRCA1/2 lwenziwe ngokwemisebenzi yezonyango kwaye zonke izigulana zinike imvume ebhaliweyo yolwazi. Izigulana nazo ziyavuma ukusetyenziswa kwedatha yazo ngeenjongo zophando.
Siyambulela uNjingalwazi uPaolo Vigneri ngoncedo lwakhe ekunyamekeleni izigulane zomhlaza webele njengoko kuceliwe yiKomiti yoBulungisa.
UFederica Martorana uxela ngembeko evela ku-Istituto Gentili, u-Eli Lilly, uNovartis, uPfizer. Abanye ababhali bathi akukho ngxabano yomdla kulo msebenzi.
1. Sung H, Ferlay J, Siegel RL, et al. Izibalo zoMhlaza weHlabathi ka-2020: I-GLOBOCAN iqikelela ukwenzeka kunye nokufa kwabantu abayi-36 abaneemhlaza kumazwe ayi-185 kwihlabathi liphela. CA Cancer J Clin.2021;71(3):209-249.doi: 10.3322/caac.21660