Gusuzuma udukoko tw’ibihaha twagaragajwe hakoreshejwe ikoranabuhanga rya "computed tomography" (CT) biracyari ikibazo mu buvuzi. Aha, turagaragaza metabolome ku isi yose y’ibipimo 480 byo mu maraso, harimo ibipimo by’ubuzima bwiza, udukoko tw’ibihaha tudakora neza, na adenocarcinoma y’ibihaha yo mu cyiciro cya mbere. Adenocarcinoma zigaragaza imiterere yihariye ya metabolome, mu gihe udukoko tw’ubuzima bwiza n’abantu bafite ubuzima bwiza bafite isano rikomeye mu miterere ya metabolome. Mu itsinda ryavumbuwe (n = 306), hagaragaye itsinda rya metabolome 27 kugira ngo hatandukanye udukoko tw’ubuzima bwiza n’utw’uburozi. AUC y’icyitegererezo cy’itandukaniro mu igenzura ry’imbere (n = 104) n’igenzura ry’inyuma (n = 111) yari 0.915 na 0.945, uko bikurikirana. Isesengura ry’inzira ryagaragaje ubwiyongere bwa metabolome za glycolytic zijyanye no kugabanuka kwa tryptophan mu maraso ya adenocarcinoma y’ibihaha ugereranije n’udukoko tw’ubuzima bwiza, kandi byagaragaje ko tryptophan ifata itera glycolysis mu turemangingo twa kanseri y’ibihaha. Ubushakashatsi bwacu bwerekanye akamaro k'ibipimo bya metabolite mu maraso mu gusuzuma ibyago byo kugaragara kw'uturemangingo tw'ibihaha na CT.
Gusuzuma hakiri kare ni ingenzi cyane kugira ngo umubare w’abarwayi ba kanseri urusheho kurokoka. Ibisubizo byavuye mu isuzuma ry’igihugu rya Amerika ryo gusuzuma kanseri y’ibihaha (NLST) n’inyigo ya NELSON y’i Burayi byagaragaje ko gusuzuma hakoreshejwe uburyo bwo gupima kanseri y’ibihaha (LDCT) buciriritse bishobora kugabanya cyane impfu za kanseri y’ibihaha mu matsinda afite ibyago byinshi1,2,3. Kuva aho LDCT ikoreshejwe cyane mu gusuzuma kanseri y’ibihaha, umubare w’ibipimo bya radiography ku buryo butunguranye byakomeje kwiyongera 4. Ibipimo bya kanseri y’ibihaha bisobanurwa nk’ibice bito bito bigera kuri cm 3 z’umurambararo 5. Duhura n’ingorane mu gusuzuma amahirwe yo kurwara kanseri no guhangana n’umubare munini w’ibipimo bya kanseri y’ibihaha byagaragaye ku buryo butunguranye kuri LDCT. Imbogamizi za CT zishobora gutuma habaho isuzuma rikurikiranwa kenshi n’ibisubizo bitari byo, bigatera ubufasha butari ngombwa no kuvurwa cyane 6. Kubwibyo, hakenewe gushyiraho ibimenyetso byizewe kandi by’ingirakamaro kugira ngo hamenyekane neza kanseri y’ibihaha mu ntangiriro no gutandukanya ibipimo byinshi bya kanseri y’ibihaha mu gihe cyo kubibona bwa mbere 7.
Isesengura ryimbitse ry’amaraso (serum, plasma, uturemangingo tw’amaraso two mu maraso), harimo na genomics, proteomics cyangwa methylation ya ADN8,9,10, byatumye abantu benshi bashishikazwa no kuvumbura ibimenyetso by’indwara ya kanseri y’ibihaha. Hagati aho, uburyo bwa metabolomic bupima umusaruro w’uturemangingo uterwa n’ibikorwa bya endogenous n’ibyo hanze, bityo bigakoreshwa mu guhanura itangira ry’indwara n’ingaruka zayo. Liquid chromatography-tandem mass spectrometry (LC-MS) ni uburyo bukoreshwa cyane mu bushakashatsi bwa metabolomics bitewe n’uburyo bunini bwo kumva no gukora ibintu bitandukanye, bushobora gupima metabolomics zifite imiterere itandukanye ya fizikiki11,12,13. Nubwo isesengura ry’imiterere ya plasma/serum ku isi ryakoreshejwe mu kumenya ibimenyetso by’indwara bifitanye isano no gusuzuma kanseri y’ibihaha14,15,16,17 n’ubushobozi bwo kuvura, 18 serum metabolite classifiers kugira ngo hamenyekane hagati y’uturemangingo tw’ibihaha tutagira ingaruka mbi n’utundi turemangingo turwaye biracyari kwigwaho cyane. -ubushakashatsi bukomeye.
Adenokasinoma na squamous cell carcinoma ni ubwoko bubiri bw'ingenzi bwa kanseri y'ibihaha itari nto (NSCLC). Ibizamini bitandukanye bya CT bigaragaza ko adenokasinoma ari yo kanseri y'ibihaha ikunze kugaragara cyane mu buryo bwa histological 1,19,20,21. Muri ubu bushakashatsi, twakoresheje ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) kugira ngo dukora isesengura rya metabolomics ku bipimo 695 bya serum, harimo ibipimo by'ubuzima, utudomo tw'ibihaha, na CT-detected ≤3 cm. Gusuzuma Stage I lung adenokasinoma. Twabonye itsinda rya metabolite mu serum zitandukanya adenokasinoma y'ibihaha n'utudomo tw'ubuzima n'ibipimo by'ubuzima. Isesengura rya Pathway ryiyongereye ryagaragaje ko tryptophan idasanzwe na glucose metabolism ari impinduka zikunze kubaho mu bipimo by'ubudomo bw'ibihaha ugereranije n'utudomo tw'ubuzima n'ibipimo by'ubuzima. Amaherezo, twashyizeho kandi twemeza uburyo bwo gushyira mu byiciro imiterere y’amaraso mu maraso bufite ubushobozi bwo gutandukanya utunyangingo tw’ibihaha twanduye n’utw’ibidakomeye twagaragaye na LDCT, bishobora gufasha mu gusuzuma hakiri kare no gusuzuma ibyago.
Mu bushakashatsi buri gukorwa, ingero z’amaraso zijyanye n’igitsina n’imyaka zakusanyijwe mu buryo bw’ibanze ku bantu 174 bazima, abarwayi 292 bafite udukoko tw’ibihaha tudakomeye, n’abarwayi 229 bafite kanseri y’ibihaha yo mu cyiciro cya mbere. Imiterere y’abaturage 695 bakoreweho ubushakashatsi igaragara mu mbonerahamwe y’inyongera ya 1.
Nkuko bigaragara ku Ishusho ya 1a, ingero 480 z'amaraso mu maraso, harimo 174 z'ubuvuzi bwiza (HC), 170 z'udukoko tudakomeye (BN), n'ingero 136 z'indwara y'ibihaha mu cyiciro cya mbere (stage I lung adenocarcinoma) mu bitaro bya Sun Yat-sen. Itsinda ry'abantu bavumbuye uburyo bwo gusuzuma imiterere y'amaraso mu maraso hakoreshejwe ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). Nkuko bigaragara ku Ishusho y'inyongera ya 1, hagaragajwe ingero zitandukanye hagati ya LA na HC, LA na BN kugira ngo hashyirweho icyitegererezo cyo gushyira mu byiciro no gukomeza gusesengura inzira zitandukanye. Ingero 104 zakusanyijwe n'Ikigo cya Kanseri cya Sun Yat-sen n'ingero 111 zakusanyijwe n'ibindi bitaro bibiri zagenzuwe imbere n'inyuma, uko bikurikirana.
a Abantu biga mu itsinda ry’abavumbuye bakorewe isesengura rya metabolomike mu maraso ku isi hose hakoreshejwe ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). b Isesengura rya "Partial least squares discriminant analysis" (PLS-DA) ry’ibipimo byose bya serum 480 byavuye mu itsinda ry’abashakashatsi, harimo n’ibipimo by’ubuzima (HC, n = 174), utudomo tudakomeye (BN, n = 170), na stage I lung adenocarcinoma (Los Angeles, n = 136). +ESI, positive electrospray ionization mode, -ESI, negative electrospray ionization mode. c–e Metabolites zifite ubwinshi butandukanye cyane mu matsinda abiri yatanzwe (ikizamini cya Wilcoxon cyashyizweho umukono n’imirizo ibiri, agaciro k’isuzuma ry’impimbano ryahinduwe, FDR <0.05) bigaragazwa mu ibara ritukura (fold change > 1.2) n’ubururu (fold change < 0.83). ) bigaragazwa ku gishushanyo cy’ikirunga. Ikarita y'ubushyuhe bw'amatsinda yerekana itandukaniro rigaragara mu mubare wa metabolite zavuzwe hagati ya LA na BN. Amakuru y'isoko atangwa mu buryo bw'amadosiye y'amakuru y'isoko.
Metabolome yose ya serum ya 174 HC, 170 BN na 136 LA mu itsinda ryavumbuwe yasesenguwe hakoreshejwe isesengura rya UPLC-HRMS. Twabanje kwerekana ko ingero zo kugenzura ubuziranenge (QC) zihurira cyane hagati mu isesengura ry’ibice by’ingenzi bitagenzuwe (PCA), byemeza ko imikorere y’ubushakashatsi buriho ubu ihamye (Ishusho y’inyongera ya 2).
Nkuko bigaragazwa mu isesengura rya partial least squares-discriminant analysis (PLS-DA) mu Ishusho ya 1 b, twasanze hari itandukaniro rigaragara hagati ya LA na BN, LA na HC mu buryo bwa electrospray ionization bwa positif (+ESI) na negative (−ESI). Ariko, nta tandukaniro rinini ryabonetse hagati ya BN na HC mu miterere ya +ESI na -ESI.
Twabonye imiterere 382 itandukanye hagati ya LA na HC, imiterere 231 itandukanye hagati ya LA na BN, n'imiterere 95 itandukanye hagati ya BN na HC (ikizamini cya Wilcoxon cyashyizweho umukono, FDR <0.05 n'impinduka nyinshi >1.2 cyangwa <0.83) (Ishusho .1c-e). . Imisozo yashyizwe ahagaragara (Amakuru y'inyongera 3) ukurikije ububiko bw'amakuru (mzCloud/HMDB/Chemspider library) ukurikije agaciro ka m/z, igihe cyo kubika no gushakisha imiterere y'amatsinda (ibisobanuro birambuye byasobanuwe mu gice cy'Uburyo) 22. Amaherezo, imiterere 33 na 38 yavuzweho ibinyabutabire bifite itandukaniro rikomeye mu bwinshi byagaragaye kuri LA ugereranije na BN (Ishusho ya 1f n'Imbonerahamwe y'inyongera 2) na LA ugereranije na HC (Ishusho y'inyongera 3 n'Imbonerahamwe y'inyongera 2), uko bikurikirana. Mu buryo bunyuranye, hagaragaye metabolite 3 gusa zifite itandukaniro rikomeye mu bwinshi muri BN na HC (Imbonerahamwe y'inyongera ya 2), ihuye n'uko BN na HC bihurirana muri PLS-DA. Izi metabolite zitandukanye zikubiyemo ubwoko bwinshi bwa biochemicals (Ishusho y'inyongera ya 4). Iyo bishyizwe hamwe, ibi byavuyemo bigaragaza impinduka zikomeye muri metabolite mu maraso igaragaza impinduka mbi ya kanseri y'ibihaha yo mu cyiciro cya mbere ugereranije n'udukoko tw'ibihaha cyangwa abantu bafite ubuzima bwiza. Hagati aho, isano ya metabolite mu maraso ya BN na HC igaragaza ko udukoko tw'ibihaha dushobora gusangira imiterere myinshi ya biyolojiya n'abantu bazima. Bitewe n'uko impinduka za gene za epidermal growth factor receptor (EGFR) zikunze kugaragara muri subtype ya 23 ya adenocarcinoma y'ibihaha, twashatse kumenya ingaruka z'impinduka za driver kuri metabolite mu maraso. Hanyuma twasesenguye imiterere rusange ya metabolite y'abantu 72 bafite EGFR mu itsinda rya adenocarcinoma y'ibihaha. Igishimishije ni uko twabonye imiterere isa hagati y’abarwayi ba EGFR mutant (n = 41) n’abarwayi ba EGFR wild-type (n = 31) mu isesengura rya PCA (Ishusho y’inyongera ya 5a). Ariko, twabonye metabolites 7 zahindutse cyane ku barwayi bafite EGFR mutation ugereranije n’abarwayi ba EGFR wild-type (ikizamini cya t, p < 0.05 na fold change > 1.2 cyangwa < 0.83) (Ishusho y’inyongera ya 5b). Inyinshi muri izi metabolites (5 kuri 7) ni acylcarnitines, zigira uruhare runini mu nzira zo gutwika aside irike.
Nkuko bigaragazwa mu mikorere igaragara ku Ishusho ya 2 a, ibipimo by'ibipimo byo gushyira mu byiciro bya nodule byabonetse hakoreshejwe uburyo bwo gutandukanya buto cyane hamwe n'uburyo bwo guhitamo hashingiwe kuri metabolite 33 zitandukanye zabonetse muri LA (n = 136) na BN (n = 170). Uburyo bwiza bwo guhuza ibipimo (LASSO) - moderi yo gusubiramo ibintu mu buryo bw'ibinyabutabire. Gusuzuma inshuro icumi byakoreshejwe mu gusuzuma uburyo bwo kwemeza ko moderi ari iy'ukuri. Guhitamo ibintu bitandukanye no kugena uburyo bwo kugena ibipimo bihindurwa hakoreshejwe igihano cyo kongera ubushobozi hamwe na parameter λ24. Isesengura rya metabolomics ku isi ryakozwe ku giti cyaryo mu matsinda yo kugenzura imbere (n = 104) no kwemeza ibintu mu buryo bwo hanze (n = 111) kugira ngo hagenzurwe imikorere y'itsinda ry'icyitegererezo cy'icyitegererezo. Kubera iyo mpamvu, metabolite 27 mu itsinda ry'ivumburwa byagaragaye ko ari moderi nziza cyane ifite agaciro ka AUC gakomeye (Ishusho ya 2b), muri yo 9 yagize ibikorwa byinshi kandi ibikorwa 18 byagabanutse muri LA ugereranije na BN (Ishusho ya 2c).
Imikorere yo kubaka uburyo bwo gushyira mu byiciro ibinure by'ibihaha, harimo guhitamo itsinda ryiza rya metabolite zo mu maraso mu itsinda ryavumbuwe hakoreshejwe icyitegererezo cya binary logistic regression binyuze mu kwemeza inshuro icumi no gusuzuma imikorere iteganyijwe mu itsinda rya verisiyo y'imbere n'iy'inyuma. b Imibare yo kwemeza icyerekezo cya LASSO regression yo guhitamo biomarker ya metabolic. Imibare yatanzwe haruguru igaragaza umubare mpuzandengo w'ibipimo byatoranijwe kuri λ runaka. Umurongo utukura w'utudomo ugaragaza agaciro mpuzandengo ka AUC kuri lambda ijyanye nayo. Utudomo tw'ibara ry'umukara tugaragaza agaciro ka AUC gato n'akatarenga. Umurongo w'utudomo ugaragaza icyitegererezo cyiza hamwe na biomarker 27 zatoranijwe. AUC, agace kari munsi y'umurongo ugaragaza imikorere ya receiver (ROC). c Impinduka za metabolite 27 zatoranijwe mu itsinda rya LA ugereranije n'itsinda rya BN mu itsinda ryavumbuwe. Inkingi itukura - gukora. Inkingi y'ubururu ni ukugabanuka. d–f Imirongo igaragaza imikorere ya receiver (ROC) igaragaza imbaraga z'icyitegererezo kigaragaza ishingiye ku mikoranire 27 ya metabolite mu itsinda ryavumbuwe, imbere, no hanze. Amakuru atangwa mu buryo bwa dosiye z'amakuru.
Icyitegererezo cyo guhanura cyakozwe hashingiwe ku bipimo byo gusubira inyuma by’izi metabolite 27 (Imbonerahamwe y’inyongera ya 3). Isesengura rya ROC rishingiye kuri izi metabolite 27 ryatanze agace kari munsi y’agaciro ka curve (AUC) ka 0.933, ubushobozi bwo kumenya itsinda ry’ivumbura bwari 0.868, naho umwihariko wari 0.859 (Ishusho ya 2d). Hagati aho, muri metabolite 38 zagaragajwe hagati ya LA na HC, itsinda rya metabolite 16 ryageze kuri AUC ya 0.902 ifite ubushobozi bwo kumenya 0.801 n’umwihariko wa 0.856 mu gutandukanya LA na HC (Ishusho y’inyongera ya 6a-c). Agaciro ka AUC gashingiye ku bipimo bitandukanye by’impinduka ku bipimo bitandukanye bya metabolite na ko kagereranyijwe. Twasanze icyitegererezo cyo gushyira mu byiciro cyakoze neza mu gutandukanya LA na BN (HC) igihe urwego rw’impinduka ku bipimo rwari rushyizwe kuri 1.2 ugereranije na 1.5 cyangwa 2.0 (Ishusho y’inyongera ya 7a,b). Icyitegererezo cyo gushyira mu byiciro, gishingiye ku matsinda 27 ya metabolite, cyarushijeho kwemezwa mu matsinda y’imbere n’iy’inyuma. AUC yari 0.915 (ubwivumbure 0.867, umwihariko 0.811) mu kwemeza imbere na 0.945 (ubwivumbure 0.810, umwihariko 0.979) mu kwemeza hanze (Ishusho ya 2e, f). Kugira ngo harebwe imikorere y’amabara atandukanye, ingero 40 zavuye mu matsinda y’inyuma zasesenguwe muri laboratwari yo hanze nk’uko byasobanuwe mu gice cy’Uburyo. Ubuziranenge bwo gushyira mu byiciro bwageze kuri AUC ya 0.925 (Ishusho y’inyongera ya 8). Kubera ko kanseri y’ibihaha ya squamous cell carcinoma (LUSC) ari yo ya kabiri ikunze kugaragara cyane muri kanseri y’ibihaha itari nto (NSCLC) nyuma ya adenocarcinoma y’ibihaha (LUAD), twanagerageje akamaro k’imikorere y’imikorere y’umubiri. BN n’abantu 16 barwaye LUSC. Ingano y’ivangura hagati ya LUSC na BN yari 0.776 (Ishusho y’inyongera ya 9), igaragaza ubushobozi buke ugereranije n’ivangura hagati ya LUAD na BN.
Ubushakashatsi bwagaragaje ko ingano y'utudomo ku mashusho ya CT ifitanye isano ryiza n'amahirwe yo kurwara kanseri kandi iracyari ikintu gikomeye kigena uburyo bwo kuvura utudomo25,26,27. Isesengura ry'amakuru yavuye mu itsinda rinini ry'ubushakashatsi bwa NELSON ryagaragaje ko ibyago byo kurwara kanseri ku bantu bafite utudomo turi munsi ya mm 5 byari bisa n'iby'abantu badafite utudomo28. Kubwibyo, ingano nto isaba gukurikiranwa buri gihe kwa CT ni mm 5, nk'uko byasabwe n'Umuryango w'Abongereza w'Ibituza (BTS), na mm 6, nk'uko byasabwe n'Umuryango wa Fleischner 29. Ariko, utudomo turenze mm 6 kandi tudafite ibimenyetso bifatika, byitwa utudomo tw'ibihaha tudasobanutse (IPN), turacyari ikibazo gikomeye mu isuzuma no gucunga mu buvuzi30,31. Twasuzumye niba ingano y'utudomo yagize ingaruka ku bimenyetso bya metabolome hakoreshejwe ingero zakusanyijwe mu matsinda yavumbuwe n'ay'abagenzura imbere. Twibanda ku bipimo 27 byemewe, twabanje kugereranya imiterere ya PCA ya metabolome ya HC na BN sub-6 mm. Twasanze amakuru menshi ya HC na BN ahuye, bigaragaza ko urugero rwa metabolite mu maraso rwari rusa mu matsinda yombi (Ishusho ya 3a). Amakarita y’imiterere mu byiciro bitandukanye by’ingano yagumye kubikwa muri BN na LA (Ishusho ya 3b, c), mu gihe hagaragaye itandukaniro hagati y’udusimba twangiza n’utwo tudakomeye mu rugero rwa mm 6-20 (Ishusho ya 3d). Iri tsinda ryari rifite AUC ya 0.927, umwihariko wa 0.868, n’ubushobozi bwo kumenya ingaruka za kanseri y’udusimba kuva kuri mm 6 kugeza kuri 20 (Ishusho ya 3e, f). Ibisubizo byacu bigaragaza ko urwego rushobora kubona impinduka mu mikorere y’umubiri ziterwa n’impinduka za kanseri mu ntangiriro, hatitawe ku bunini bw’udusimba.
Igereranya ry'imiterere ya PCA hagati y'amatsinda yagenwe hashingiwe ku gipimo cya metabolike cya metabolike 27. CC na BN < 6 mm. b BN < 6 mm ugereranije na BN 6–20 mm. muri LA 6–20 mm ugereranije na LA 20–30 mm. g BN 6–20 mm na LA 6–20 mm. GC, n = 174; BN < 6 mm, n = 153; BN 6–20 mm, n = 91; LA 6–20 mm, n = 89; LA 20–30 mm, n = 77. e Umurongo ugaragaza imikorere ya Receiver operating (ROC) ugaragaza imikorere itandukanye ya moderi ku tudomo twa mm 6–20. f Agaciro k'ubushobozi kabazwe hashingiwe ku gipimo cya logistic regression kuri tudomo twa mm 6–20. Umurongo w'imvi ugaragaza agaciro ka cutoff (0.455). Imibare iri hejuru igaragaza ijanisha ry'imanza ziteganijwe muri Los Angeles. Koresha ikizamini cya t cy’umunyeshuri gifite imirizo ibiri. PCA, isesengura ry’ibice by’ingenzi. Agace ka AUC kari munsi y’umurongo. Amakuru y’isoko atangwa mu buryo bwa dosiye z’amakuru y’isoko.
Ingero enye (z'imyaka 44-61) zifite ingano isa y'utudomo tw'ibihaha (7-9 mm) zatoranyijwe kugira ngo zigaragaze imikorere y'icyitegererezo cyatanzwe cyo guhanura kanseri (Ishusho ya 4a, b). Mu isuzuma rya mbere, Urubanza rwa 1 rwagaragaye nk'utudomo dukomeye dufite calcium, ikintu gifitanye isano n'ubudahangarwa, mu gihe Urubanza rwa 2 rwagaragaye nk'utudomo dukomeye duto tudafite ibimenyetso bigaragara by'ubudahangarwa. Inshuro eshatu zo gupima CT zakurikiranye zagaragaje ko izi ndwara zakomeje kuba nziza mu gihe cy'imyaka 4, bityo zifatwa nk'utudomo tudakomeye (Ishusho ya 4a). Ugereranije n'isuzuma rya muganga rya CT scans, isesengura rya metabolite y'amaraso rimwe hamwe n'icyitegererezo cy'ubu buryo bwo gupima indwara bwagaragaje vuba kandi neza izi tudomo duto hashingiwe ku mbogamizi zishobora kubaho (Imbonerahamwe ya 1). Ishusho ya 4b mu rubanza rwa 3 igaragaza akadomo gafite ibimenyetso byo gutakaza pleural, akenshi gafitanye isano na kanseri32. Urubanza rwa 4 rwagaragaye nk'utudomo dukomeye duto tudafite ibimenyetso by'impamvu idakomeye. Izi ndwara zose zavuzwe ko ari mbi hakurikijwe icyitegererezo cy'itsinda (Imbonerahamwe ya 1). Isuzuma rya adenocarcinoma y'ibihaha ryagaragajwe n'isuzuma ry'indwara nyuma yo kubagwa ibihaha (Ishusho ya 4b). Ku isuzuma ry'inyuma, umuhanga mu gusesengura imikorere y'umubiri yavuze neza ko hari udukoko tubiri tw'ibihaha tudafite uburebure burenga mm 6 (Ishusho y'inyongera ya 10).
Amashusho ya CT y'idirishya ry'imirongo y'ibihaha ry'abantu babiri bafite utudomo tudakomeye. Mu rubanza rwa 1, CT scan nyuma y'imyaka 4 yagaragaje akadomo gakomeye gafite uburebure bwa mm 7 hamwe na calciumification mu gice cyo hasi cy'iburyo. Mu rubanza rwa 2, CT scan nyuma y'imyaka 5 yagaragaje akadomo gakomeye gafite umurambararo wa mm 7 mu gice cyo hejuru cy'iburyo. b Amashusho ya CT y'idirishya ry'imirongo n'inyigo zijyanye n'indwara z'abantu babiri barwaye kanseri y'impatwe mbere yo gucibwa ibihaha. Urubanza rwa 3 rwagaragaje akadomo gafite umurambararo wa mm 8 mu gice cyo hejuru cy'iburyo hamwe no gukurura pleural. Urubanza rwa 4 rwagaragaje akadomo gakomeye gafite uburebure bwa mm 9 mu gice cyo hejuru cy'ibumoso. Hematoxylin na eosin (H&E) ibara ry'ibihaha byakuweho (urugero = 50 μm) bigaragaza imiterere y'ikura rya kanseri y'ibihaha. Imyambi igaragaza utudomo twabonetse ku mashusho ya CT. Amashusho ya H&E ni amashusho agaragaza imirima myinshi (>3) mikorobe yasuzumwe n'umuganga w'indwara.
Ibyo twakoze byose hamwe bigaragaza akamaro gashoboka k’ibipimo bya metabolite mu maraso mu gusuzuma udukoko tw’ibihaha, ibyo bikaba bishobora guteza ibibazo mu gusuzuma isuzuma rya CT.
Dushingiye ku gice cy’isesengura ry’imiterere y’umubiri cyemewe, twashatse kumenya isano iri hagati y’impinduka zikomeye mu mikorere y’umubiri. Isesengura rya KEGG ryakozwe na MetaboAnalyst ryagaragaje inzira 6 zisanzwe zahindutse cyane hagati y’amatsinda abiri yatanzwe (LA vs. HC na LA vs. BN, ryahinduwe p ≤ 0.001, ingaruka > 0.01). Izi mpinduka zaranzwe n’ihungabana mu mikorere y’umubiri ya pyruvate, imikorere ya tryptophan, imikorere ya niacin na nicotinamide, glycolysis, TCA cycle, na purine metabolism (Ishusho ya 5a). Hanyuma twakoze isuzuma rya metabolomics rigamije kwemeza impinduka zikomeye dukoresheje isuzuma ry’imiterere y’umubiri. Kumenya metabolomics zisanzwe mu nzira zisanzwe zahinduwe hakoreshejwe spectrometry ya triple quadrupole mass (QQQ) dukoresheje amahame nyayo ya metabolite. Imiterere y'abaturage mu bushakashatsi bwa metabolomics ikubiye mu mbonerahamwe y'inyongera ya 4. Dukurikije ibyavuye mu bushakashatsi ku isi, isesengura ry'imibare ryemeje ko hypoxanthine na xanthine, pyruvate, na lactate byiyongereye muri LA ugereranije na BN na HC (Ishusho ya 5b, c, p <0.05). Ariko, nta tandukaniro rigaragara muri izi metabolomics ryabonetse hagati ya BN na HC.
Isesengura rya KEGG ry’uburyo bwo kongera ubushobozi bw’umubiri mu itsinda rya LA ugereranije n’amatsinda ya BN na HC. Hakoreshejwe ikizamini cya Globaltest gifite imirizo ibiri, maze agaciro ka p karahindurwa hakoreshejwe uburyo bwa Holm-Bonferroni (p yahinduwe ≤ 0.001 n’ingano y’ingaruka > 0.01). b–d Ibishushanyo bya Violin bigaragaza urugero rwa hypoxanthine, xanthine, lactate, pyruvate, na tryptophan muri serum HC, BN, na LA bigenwa na LC-MS/MS (n = 70 kuri buri tsinda). Imirongo y’utudomo tw’umweru n’umukara igaragaza imiterere y’igipimo cya median na quartile, uko bikurikirana. e Ishusho ya Violin igaragaza Log2TPM (inyandiko kuri miliyoni) ya mRNA ya SLC7A5 na QPRT muri adenocarcinoma y’ibihaha (n = 513) ugereranije n’ingingo z’ibihaha zisanzwe (n = 59) muri LUAD-TCGA dataset. Agasanduku k'umweru kagaragaza urwego rwa interquartile, umurongo w'umukara utambitse hagati ugaragaza urwego rw'igipimo, naho umurongo w'umukara uhagaze uva mu gasanduku ugaragaza igihe cy'icyizere cya 95% (CI). f Igishushanyo cya Pearson cy'itandukaniro rya SLC7A5 na GAPDH mu ndwara y'ibihaha (n = 513) n'ingingo zisanzwe z'ibihaha (n = 59) mu makuru ya TCGA. Agace k'umukara kagaragaza 95% CI. r, coefficient ya Pearson. g Ingano ya tryptophan isanzwe mu turemangingo twa A549 yahinduwe hakoreshejwe uburyo butari bwihariye bwo kugenzura shRNA (NC) na shSLC7A5 (Sh1, Sh2) byagenwe na LC-MS/MS. Isesengura ry'ingero eshanu zigenga ibinyabuzima muri buri tsinda riragaragazwa. h Ingano ya NADt mu turemangingo (NAD yose, harimo na NAD+ na NADH) mu turemangingo twa A549 (NC) na SLC7A5 knockdown A549 (Sh1, Sh2). Isesengura ry'ingero eshatu zigenga ibinyabuzima muri buri tsinda riragaragazwa. Imikorere ya glycolytic y’uturemangingo twa A549 mbere na nyuma yo gukubita SLC7A5 yapimwe hakoreshejwe igipimo cya aside extracellular (ECAR) (n = ingero 4 zigenga ibinyabuzima kuri buri tsinda). 2-DG, 2-deoxy-D-glucose. Ikizamini cya t cy’umunyeshuri ufite imirizo ibiri cyakoreshejwe muri (b–h). Muri (g–i), utubari tw’amakosa tugaragaza impuzandengo ya ± SD, buri gerageza ryakozwe inshuro eshatu ku giti cyaryo kandi ibisubizo byari bisa. Amakuru y’isoko atangwa mu buryo bwa dosiye y’isoko.
Dukurikije ingaruka zikomeye z’impinduka mu mikorere ya tryptophan mu itsinda rya LA, twanasuzumye urugero rwa tryptophan mu maraso mu matsinda ya HC, BN, na LA dukoresheje QQQ. Twasanze tryptophan mu maraso yagabanutse muri LA ugereranije na HC cyangwa BN (p < 0.001, Ishusho ya 5d), ibi bikaba bihuye n’ibyavumbuwe mbere ko urugero rwa tryptophan ruzenguruka mu maraso ari hasi mu barwayi ba kanseri y’ibihaha ugereranyije n’abashinzwe kugenzura ubuzima bwiza bo mu itsinda ryagenzuwe33,34,35. Ubundi bushakashatsi bwakoresheje PET/CT tracer 11C-methyl-L-tryptophan bwagaragaje ko igihe cyo kubika ikimenyetso cya tryptophan mu maraso y’ibihaha cyiyongereye cyane ugereranije n’ibisebe bitagira ingaruka mbi cyangwa ingingo zisanzwe36. Dutekereza ko kugabanuka kwa tryptophan mu maraso ya LA bishobora kugaragaza ko uturemangingo twa kanseri y’ibihaha twinjira muri tryptophan.
Birazwi kandi ko umusaruro wa nyuma w'inzira ya kynurenine ya tryptophan catabolism ari NAD+37,38, ikaba ari substrate y'ingenzi mu gukora kwa glyceraldehyde-3-phosphate hamwe na 1,3-bisphosphoglycerate muri glycolysis39. Nubwo ubushakashatsi bwabanje bwibanze ku ruhare rwa tryptophan catabolism mu mikorere y'ubudahangarwa bw'umubiri, twashatse gusobanura imikoranire iri hagati ya tryptophan dysregulation n'inzira za glycolytic zagaragaye muri ubu bushakashatsi. Solute transporter family 7 member 5 (SLC7A5) izwiho kuba tryptophan transporter43,44,45. Quinolinic acid phosphoribosyltransferase (QPRT) ni enzyme iri munsi y'inzira ya kynurenine ihindura aside quinolinic muri NAMN46. Isuzuma ry'amakuru ya LUAD TCGA ryagaragaje ko SLC7A5 na QPRT byombi byazamutse cyane mu tunyangingo twa kanseri ugereranije n'utunyangingo dusanzwe (Ishusho ya 5e). Uku kwiyongera kwagaragaye mu cyiciro cya mbere n'icya kabiri ndetse no mu cyiciro cya gatatu n'icya kane cy'indwara y'ibihaha (Ishusho y'inyongera ya 11), bigaragaza ko hakiri kare habayeho ibibazo mu mikorere ya tryptophan bifitanye isano no kubyara kanseri.
Byongeye kandi, amakuru ya LUAD-TCGA yagaragaje isano iri hagati ya SLC7A5 na GAPDH mRNA mu ngero z'abarwayi ba kanseri (r = 0.45, p = 1.55E-26, Ishusho ya 5f). Mu buryo bunyuranye, nta sano igaragara yabonetse hagati y'ibimenyetso nk'ibi bya gene mu ngingo zisanzwe z'ibihaha (r = 0.25, p = 0.06, Ishusho ya 5f). Gutakaza SLC7A5 (Ishusho y'inyongera ya 12) mu turemangingo twa A549 byagabanyije cyane urugero rwa tryptophan na NAD (H) mu turemangingo (Ishusho ya 5g, h), bigatuma imikorere ya glycolytic igabanuka nk'uko bipimwa n'igipimo cya aside extracellular (ECAR) (Ishusho ya 1). 5i). Bityo, dushingiye ku mpinduka mu mikorere ya metabolic mu maraso no mu gupima muri vitro, dutekereza ko tryptophan metabolism ishobora gukora NAD+ binyuze mu nzira ya kynurenine kandi ikagira uruhare runini mu guteza imbere glycolysis muri kanseri y'ibihaha.
Ubushakashatsi bwagaragaje ko umubare munini w'uturemangingo tw'ibihaha tutazwi twabonetse na LDCT dushobora gutuma hakenerwa ibizamini by'inyongera nka PET-CT, biopsy y'ibihaha, no kuvurwa cyane bitewe n'isuzuma ry'indwara ya kanseri.31 Nkuko bigaragara ku Ishusho ya 6, ubushakashatsi bwacu bwagaragaje itsinda ry'ibintu bishobora gupima mu maraso bifite agaciro gashoboka ko byakwitabwaho bishobora kunoza uburyo bwo gutandukanya ingaruka no gucunga uturemangingo tw'ibihaha twabonetse na CT.
Udukoko tw’ibihaha dupimwa hakoreshejwe uburyo bwo gupima indwara (LDCT) bufite ubushobozi bwo gupima indwara bugaragaza impamvu zitari mbi cyangwa mbi. Ingaruka zitazwi z’utu tukoko zishobora gutuma umuntu akurikiranwa kenshi, agafatwa ingamba zitari ngombwa, ndetse akanavurwa cyane. Gushyiramo imiti igabanya ubukana bw’amaraso mu maraso ifite agaciro ko gusuzuma indwara bishobora kunoza isuzuma ry’ibyago no gucunga udukoko tw’ibihaha nyuma yaho. PET positron emission tomography.
Amakuru yavuye mu bushakashatsi bwa NLST bwo muri Amerika n'ubushakashatsi bwa NELSON bw'i Burayi agaragaza ko gusuzuma amatsinda afite ibyago byinshi hakoreshejwe uburyo bwo gupima bwa LDCT (LDCT) bukoresha dose nkeya bishobora kugabanya impfu za kanseri y'ibihaha1,3. Ariko, gusuzuma ibyago no gukurikirana nyuma y'aho umubare munini w'uturemangingo tw'ibihaha twagaragaye na LDCT biracyari ingorabahizi cyane. Intego nyamukuru ni ugushyira mu byiciro bikwiye protocole zishingiye kuri LDCT zisanzweho hifashishijwe biomarkers yizewe.
Bimwe mu bipimo bya molekile, nka metabolites zo mu maraso, byagaragaye hagaragazwa kanseri y'ibihaha n'ibipimo by'ubuzima bwiza15,17. Mu bushakashatsi buriho, twibanze ku ikoreshwa ry'isesengura rya metabolites zo mu maraso kugira ngo dutandukanye utudomo tw'ibihaha tudahumanye n'utw'indwara mbi twagaragaye mu buryo butunguranye na LDCT. Twagereranije metabolites zo mu maraso ku isi yose zo mu maraso zigenzura ubuzima bwiza (HC), utudomo tw'ibihaha tudakomeye (BN), n'ibipimo bya adenocarcinoma y'ibihaha ku cyiciro cya mbere (LA) dukoresheje isesengura rya UPLC-HRMS. Twasanze HC na BN bifite imiterere isa ya metabolites, mu gihe LA yagaragaje impinduka zikomeye ugereranije na HC na BN. Twasanze ibice bibiri bya metabolites zo mu maraso zitandukanya LA na HC na BN.
Gahunda yo kumenya udukoko tudakomeye n’utubi duto ishingiye kuri LDCT ishingiye ahanini ku bunini, ubucucike, imiterere n’umuvuduko w’udukoko uko igihe kigenda gihita30. Ubushakashatsi bwakozwe mbere bwagaragaje ko ingano y’udukoko ifitanye isano rya hafi n’amahirwe yo kurwara kanseri y’ibihaha. Ndetse no ku barwayi bafite ibyago byinshi byo kurwara kanseri mu dukoko turi munsi ya mm 6 ni <1%. Ingaruka zo kurwara kanseri ku dukoko dufite uburebure bwa mm 6 kugeza kuri 20 ziri hagati ya 8% na 64% na 30. Kubwibyo, Umuryango wa Fleischner usaba ko umurambararo wa mm 6 ushyirwa mu byiciro kugira ngo hakurikiranwe CT buri gihe. 29 Ariko, isuzuma n’imicungire y’udukoko duto tw’ibihaha (IPN) turenze mm 6 ntirakorwa neza 31. Ubusanzwe ubuvuzi bw’indwara z’umutima busanzwe bushingiye ku gutegereza witonze hamwe no gukurikirana CT kenshi.
Dushingiye kuri metabolome yemewe, twerekanye ku nshuro ya mbere isano y’ibimenyetso bya metabolome hagati y’abantu bazima n’udusimba tudafite ingaruka nziza <6 mm. Ukuri ku binyabuzima bihuye n’ibyavumbuwe mbere na CT ko ibyago byo kurwara kanseri y’udusimba tudafite ingaruka nziza ari bike nk’uko bimeze ku bantu badafite udusimba.30 Byagombye kumenyekana ko ibyavuye mu bushakashatsi bwacu bigaragaza ko udusimba tudafite ingaruka nziza <6 mm na ≥6 mm dufite isano ikomeye mu miterere ya metabolome, bigaragaza ko ibisobanuro by’imikorere y’impamvu ya tekiniki idahinduka hatitawe ku bunini bw’udusimba. Bityo, ibipimo bya metabolite ya serum igezweho bishobora gutanga ikizamini kimwe nk’ikizamini cyo kudakora iyo udusimba tubonetse bwa mbere kuri CT kandi bishobora kugabanya gukurikirana inshuro nyinshi. Muri icyo gihe, itsinda rimwe ry’ibipimo bya metabolike ryatandukanije utusimba tudafite ingaruka nziza dufite ≥6 mm n’udusimba tudafite ingaruka nziza kandi ryatanze ibizamini nyabyo bya IPN zingana n’imiterere idasobanutse neza ku mashusho ya CT. Iyi serum metabolism classifier yitwaye neza mu guhanura kanseri y’udusimba ≥6 mm hamwe na AUC ya 0.927. Ubushakashatsi bwacu bwerekana ko ibimenyetso byihariye bya metabolomike mu maraso bishobora kugaragaza by’umwihariko impinduka za mbere mu mikorere y’umubiri ziterwa n’ikibyimba kandi bikaba bifite akamaro nk’ibigaragaza ibyago, hatitawe ku bunini bw’uturemangingo.
Ikigaragara ni uko kanseri y’ibihaha (LUAD) na kanseri y’uturemangingo twa squamous cell (LUSC) ari byo bwoko bw’ingenzi bwa kanseri y’ibihaha itari nto (NSCLC). Bitewe n’uko LUSC ifitanye isano rikomeye n’ikoreshwa ry’itabi47 kandi LUAD ikaba ari yo ikunze kugaragara cyane mu turemangingo tw’ibihaha twabonetse mu gusuzuma CT48, icyitegererezo cyacu cyo mu cyiciro cya mbere cy’uturemangingo cyakozwe by’umwihariko ku bipimo bya adenocarcinoma yo mu cyiciro cya mbere. Wang na bagenzi be kandi bibanze kuri LUAD maze bamenya ibimenyetso icyenda bya lipide bakoresheje lipidomike kugira ngo batandukanye kanseri y’ibihaha yo mu cyiciro cya mbere n’abantu bazima17. Twagerageje icyitegererezo cy’ubu cy’uturemangingo ku bantu 16 ba LUSC yo mu cyiciro cya mbere n’uturemangingo 74 tudakomeye kandi twabonye ko LUSC yari ifite ubushobozi bwo guhanura neza (AUC 0.776), bigaragaza ko LUAD na LUSC zishobora kuba zifite ibimenyetso byazo bwite bya metabolomike. Mu by’ukuri, LUAD na LUSC byagaragaye ko bitandukanye mu mpamvu zikomoka ku ndwara, inkomoko y’ibinyabuzima n’impinduka mu miterere y’uturemangingo49. Kubwibyo, ubundi bwoko bwa histology bugomba gushyirwa mu mahugurwa yo gupima kanseri y'ibihaha hifashishijwe abaturage muri gahunda zo gupima.
Aha, twabonye inzira esheshatu zikunze guhinduka cyane muri adenocarcinoma y'ibihaha ugereranije n'izigenzura ubuzima bwiza n'udusimba tudakomeye. Xanthine na hypoxanthine ni metabolite zisanzwe mu nzira ya purine metabolism. Dukurikije ibyavuye mu bushakashatsi bwacu, ibice bifitanye isano na purine metabolism byiyongereye cyane mu maraso cyangwa mu ngingo z'abarwayi bafite adenocarcinoma y'ibihaha ugereranije n'abagenzura ubuzima bwiza cyangwa abarwayi bari mu cyiciro cya mbere cyo kuva mu maraso15,50. Urugero rwo hejuru rwa xanthine na hypoxanthine mu maraso rushobora kugaragaza anabolism isabwa n'uturemangingo twa kanseri twiyongera vuba. Kugabanuka kwa glucose metabolism ni ikimenyetso kizwi cyane cy'isesengura rya kanseri51. Aha, twabonye ubwiyongere bunini bwa pyruvate na lactate mu itsinda rya LA ugereranije n'itsinda rya HC na BN, ibi bikaba bihuye n'ibyatangajwe mbere ku buryo glycolytic pathway idakora neza mu maraso ya metabolome mu maraso y'abarwayi ba kanseri y'ibihaha itari nto (NSCLC) n'abagenzura ubuzima bwiza. Ibisubizo birahuye52,53.
Icy'ingenzi ni uko twabonye isano iri hagati ya metabolism ya pyruvate na tryptophan mu maraso y'ibihaha. Igipimo cya tryptophan mu maraso cyagabanutse mu itsinda rya LA ugereranije n'itsinda rya HC cyangwa BN. Igishimishije ni uko ubushakashatsi bwakozwe mbere bwakoresheje itsinda ry'abantu bateganya ko tryptophan nke zigenda zikwirakwira mu maraso bwagaragaje ko urugero ruto rwa tryptophan rugenda rutera ibyago byinshi byo kurwara kanseri y'ibihaha. Tryptophan ni aside amine y'ingenzi tubona mu biribwa gusa. Twanzuye ko kugabanuka kwa tryptophan mu maraso y'ibihaha bishobora kugaragaza kugabanuka vuba kw'iyi metabolite. Birazwi neza ko umusaruro wa tryptophan catabolism binyuze mu nzira ya kynurenine ari wo soko y'ikorwa rya NAD+ nshya. Kubera ko NAD+ ikorwa ahanini binyuze mu nzira yo gukiza, akamaro ka NAD+ mu metabolite ya tryptophan mu buzima no mu ndwara karacyari ngombwa kumenyekana46. Isesengura ryacu rya TCGA ryagaragaje ko imikorere ya tryptophan transporter solute transporter 7A5 (SLC7A5) yiyongereye cyane muri adenocarcinoma y'ibihaha ugereranije n'uburyo busanzwe bwo kugenzura kandi byari bifitanye isano ryiza n'imikorere ya enzyme ya glycolytic GAPDH. Ubushakashatsi bwabanje bwibanze ahanini ku ruhare rwa tryptophan catabolism mu guhagarika ubudahangarwa bw'umubiri bwo kurwanya kanseri40,41,42. Aha tugaragaza ko gukumira tryptophan ikwirakwira no guhosha SLC7A5 mu turemangingo twa kanseri y'ibihaha bituma urugero rwa NAD mu turemangingo rugabanuka ndetse no kugabanya imikorere ya glycolytic. Muri make, ubushakashatsi bwacu butanga ishingiro ry'ibinyabuzima ku mpinduka mu mikorere y'amaraso zijyanye n'impinduka mbi za adenocarcinoma y'ibihaha.
Impinduka za EGFR ni zo zikunze gutera impinduka mu barwayi ba NSCLC. Mu bushakashatsi bwacu, twasanze abarwayi bafite impinduka za EGFR (n = 41) bari bafite imiterere ya metabolome muri rusange isa n’iy’abarwayi bafite EGFR yo mu bwoko bwa wild-type (n = 31), nubwo twasanze urugero rwa serum rwagabanutse rwa bamwe mu barwayi ba EGFR mutant mu barwayi ba acylcarnitine mu maraso. Inshingano ya acylcarnitines ni ugutwara amatsinda ya acyl kuva kuri cytoplasme ikajya muri mitochondrial matrix, bigatuma aside fatty acids ikora ingufu 55. Dukurikije ibyo twabonye, ubushakashatsi buherutse kandi bwagaragaje imiterere ya metabolome isa hagati ya EGFR mutant na EGFR wild-type tumors isesengura metabolome yo ku isi yose y’ibice 102 by’ibihaha bya adenocarcinoma50. Igishimishije ni uko ingano ya acylcarnitine nayo yabonetse mu itsinda rya EGFR mutant. Kubwibyo, niba impinduka mu rugero rwa acylcarnitine zigaragaza impinduka mu imikorere ya EGFR n’inzira za molekile zishobora kuba zikenewe kwigwaho byinshi.
Mu gusoza, ubushakashatsi bwacu bwashyizeho uburyo bwo gupima imikorere y’amaraso mu maraso kugira ngo hamenyekane itandukaniro ry’uturemangingo tw’ibihaha, kandi butanga uburyo bwo gukora bushobora kunoza isuzuma ry’ibyago no koroshya imicungire y’indwara hashingiwe ku gusuzuma CT scan.
Ubu bushakashatsi bwemejwe na Komite Ishinzwe Imyitwarire y’Ibitaro bya Kanseri bya Kaminuza ya Sun Yat-sen, Ibitaro bya Mbere bifitanye isano bya Kaminuza ya Sun Yat-sen, na Komite Ishinzwe Imyitwarire y’Ibitaro bya Kanseri bya Kaminuza ya Zhengzhou. Mu matsinda yavumbuwe n’ay’igenzura ry’imbere mu mubiri, hatoranijwe abantu 174 barwaye indwara z’imitsi n’abantu 244 barwaye indwara z’imitsi n ... Ikigo cya Kaminuza ya Sun Yat-sen Cancer Center cyakusanyije kandi abantu 16 barwaye kanseri y’ibihaha yo mu cyiciro cya mbere kugira ngo bapime ubushobozi bwo gusuzuma imiterere y’umubiri (imiterere y’umurwayi igaragara mu mbonerahamwe y’inyongera ya 5). Ingero z’itsinda ry’abavumbuye n’itsinda ry’abavumbuye zakusanyijwe hagati ya Mutarama 2018 na Gicurasi 2020. Ingero z’itsinda ry’abavumbuye zakusanyijwe hagati ya Kanama 2021 na Ukwakira 2022. Kugira ngo bigabanye ivangura rishingiye ku gitsina, umubare w’abagabo n’abagore ungana washyizwe kuri buri tsinda. Itsinda ry’abavumbuye n’itsinda ry’abasuzuma imbere. Igitsina cy’abitabiriye cyagenwe hashingiwe ku raporo y’umuntu ku giti cye. Ubwemeracyaha bwatanzwe n’abitabiriye bose kandi nta ndishyi zatanzwe. Abarwaye kanseri y’ibihaha ni abafite amanota ahamye ya CT scan ku myaka 2 kugeza kuri 5 mu gihe cy’isesengura, uretse umuntu 1 wavuye mu cyiciro cy’abavumbuye inyuma, cyakusanyijwe mbere yo kubagwa no gupimwa histopathology. Uretse indwara ya bronchitis idakira. Abarwayi ba adenocarcinoma y’ibihaha bakusanyijwe mbere yo gucibwa ibihaha kandi byemezwa n’indwara. Ingero z'amaraso yo kwiyiriza ubusa zakusanyijwe mu miyoboro yo gutandukanya serum nta miti igabanya amaraso. Ingero z'amaraso zafashwe isaha imwe ku bushyuhe bw'icyumba hanyuma zishyirwa kuri centrifuge kuri 2851 × g mu gihe cy'iminota 10 kuri 4°C kugira ngo hakusanywe serum supernatant. Ingano z'amaraso ya serum zashyizwe muri -80°C kugeza igihe hakuwemo metabolite. Ishami rishinzwe gukumira kanseri n'isuzuma ry'ubuvuzi ry'ikigo cya kaminuza cya Sun Yat-sen Cancer Center ryakusanyije serum yavuye mu batanga serum 100 bazima, barimo abagabo n'abagore bari hagati y'imyaka 40 na 55. Ingano zingana za buri ngero z'abatanga serum zaravanzwe, serum yavuyemo irakurwaho irabikwa kuri -80°C. Uruvange rw'amaraso rwakoreshejwe nk'igikoresho cyo kugenzura ubuziranenge no gupima amakuru.
Ingero za serum n'ingero z'ibizamini byarashongeshejwe, hanyuma hakurwamo ibintu bya metabolite hakoreshejwe uburyo bwo gukurura (MTBE/methanol/amazi) 56. Muri make, 50 μl za serum zavanzwe na 225 μl za methanol ikonje cyane na 750 μl za methyl tert-butyl ether ikonje cyane (MTBE). Kangura uruvange hanyuma ushyire ku rubura isaha 1. Ingero zavanzwe hanyuma vortex ivangwa na 188 μl z'amazi yo mu rwego rwa MS arimo ibipimo by'imbere (13C-lactate, 13C3-pyruvate, 13C-methionine, na 13C6-isoleucine, byaguzwe muri Laboratories za Cambridge Isotope). Hanyuma uruvange rwashyizwe kuri centrifuge kuri 15.000 × g mu gihe cy'iminota 10 kuri 4 °C, hanyuma igice cyo hasi cyimurirwa mu miyoboro ibiri (125 μL buri imwe) kugira ngo hasesengurwe LC-MS mu buryo bwiza n'ububi. Amaherezo, icyitegererezo cyarahindutse umwuka kikamera mu cyuma gikonjesha umwuka cyihuta cyane.
Ibinyabutabire byumye byongeye gushyirwa mu mililitiro 120 za acetonitrile ya 80%, bishyirwa mu mwobo mu gihe cy'iminota 5, hanyuma bishyirwa mu buryo bwa centrifuge kuri 15.000 × g mu gihe cy'iminota 10 kuri 4°C. Supernatants zoherejwe mu ducupa tw'ikirahure cy'umuhondo hamwe n'ibikoresho bito kugira ngo bikoreshwe mu bushakashatsi bwa metabolomics. Isesengura rya metabolomics ritagamije intego ku buryo bwa ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). Metabolites zatandukanyijwe hakoreshejwe sisitemu ya Dionex Ultimate 3000 UPLC na ACQUITY BEH Amide column (2.1 × 100 mm, 1.7 μm, Waters). Mu buryo bwa positive ion, ibyiciro byo kugenda byari 95% (A) na 50% acetonitrile (B), buri kimwe kirimo 10 mmol / L ammonium acetate na 0.1% formic acid. Mu buryo bwa negative, ibyiciro bya mobile phase A na B byari bifite 95% na 50% acetonitrile, uko bikurikirana, ibyiciro byombi byari bifite 10 mmol/L ammonium acetate, pH = 9. Gahunda ya gradient yari ikurikira: 0–0.5 min, 2% B; 0.5–12 min, 2–50% B; 12–14 min, 50–98% B; 14–16 min, 98% B; 16–16.1 min, 98–2% B; 16.1–20 min, 2% B. Inkingi yagumye kuri 40°C naho icyitegererezo kuri 10°C muri autosampler. Igipimo cy'amazi cyari 0.3 ml/min, ingano y'igishinge yari 3 μl. Spektrometero y’ubusa ya Q-Exactive Orbitrap (Thermo Fisher Scientific) ifite isoko ya electrospray ionization (ESI) yakoreshejwe mu buryo bwa scan yuzuye kandi ihuzwa n’uburyo bwo gukurikirana bwa ddMS2 kugira ngo hakusanywe amakuru menshi. Ibipimo bya MS byashyizweho ku buryo bukurikira: voltage ya spray +3.8 kV/- 3.2 kV, ubushyuhe bwa capillary 320°C, gaze yo kurinda 40 arb, gaze yunganira 10 arb, ubushyuhe bwa probe heater 350°C, intera yo gushakisha 70–1050 m/h, resolution. 70 000. Amakuru yabonetse hakoreshejwe Xcalibur 4.1 (Thermo Fisher Scientific).
Kugira ngo hasuzumwe ubuziranenge bw'amakuru, ingero zashyizwe hamwe zo kugenzura ubuziranenge (QC) zakozwe hakurwamo 10 μL ya supernatant muri buri cyitegererezo. Ingero esheshatu zo kugenzura ubuziranenge zasesenguwe mu ntangiriro y'uruhererekane rw'isesengura kugira ngo hamenyekane ubuziranenge bw'uburyo bwa UPLC-MS. Ingero zo kugenzura ubuziranenge zishyirwa mu itsinda rimwe na rimwe. Ingero zose 11 z'ingero za serum muri ubu bushakashatsi zasesenguwe na LC-MS. Ingero z'imvange ya serum pool y'abantu 100 bazima zakoreshejwe nk'ibikoresho byo kwifashisha mu byiciro bitandukanye kugira ngo hagenzurwe uburyo bwo gukuramo no guhindura ingaruka za batch-to-batch. Isesengura rya metabolomics ritagamije kugaragara ku itsinda ry'abavumbuye, itsinda ry'abagenzura imbere, n'itsinda ry'abagenzura hanze ryakorewe mu Kigo cya Metabolomics cya Kaminuza ya Sun Yat-sen. Laboratwari yo hanze ya Kaminuza ya Guangdong y'Ikoranabuhanga mu Isesengura n'Ibizamini nayo yasesenguye ingero 40 zo hanze kugira ngo isuzume imikorere y'icyitegererezo cy'isesengura.
Nyuma yo gukuramo no kongera gushyira mu bikorwa, hapimwe umubare wuzuye wa metabolite zo mu maraso hakoreshejwe uburyo bwa ultra-performance high liquid chromatography-tandem mass spectrometry (Agilent 6495 triple quadrupole) hamwe n'isoko rya electrospray ionization (ESI) muri mode multiple reaction monitoring (MRM). Hakoreshejwe ACQUITY BEH Amide column (2.1 × 100 mm, 1.7 μm, Waters) mu gutandukanya metabolite. Icyiciro cyo kugenda cyari kigizwe na 90% (A) na 5% acetonitrile (B) hamwe na 10 mmol/L ammonium acetate na 0.1% ammonia solution. Gahunda ya gradient yari iyi ikurikira: 0–1.5 min, 0% B; 1.5–6.5 min, 0–15% B; 6.5–8 min, 15% B; 8–8.5 min, 15%–0% B; 8.5–11.5 min, 0% B. Inkingi yagumanye ubushyuhe kuri 40 °C n'icyitegererezo kuri 10 °C muri autosampler. Igipimo cy'amazi cyari 0.3 mL/min naho ingano y'ingufu yari 1 μL. Ibipimo bya MS byashyizweho ku buryo bukurikira: voltage ya capillary ± 3.5 kV, umuvuduko wa nebulizer 35 psi, sheath gaz flow 12 L/min, sheath gaz flow 350 °C, suspension gaz flow 250 °C, na suspension gaz flow 14 l/min. Impinduka za MRM za tryptophan, pyruvate, lactate, hypoxanthine na xanthine zari 205.0–187.9, 87.0–43.4, 89.0–43.3, 135.0–92.3 na 151.0–107. 9 uko bikurikirana. Amakuru yakusanyijwe hakoreshejwe Mass Hunter B.07.00 (Agilent Technologies). Ku bipimo byo mu maraso, tryptophan, pyruvate, lactate, hypoxanthine, na xanthine byapimwe hakoreshejwe uburyo bwo gupima imvange isanzwe. Ku bipimo by'uturemangingo, ingano ya tryptophan yagereranyijwe n'ingano y'imbere mu mubiri n'ingano ya poroteyine mu turemangingo.
Gukuramo ibipimo by’umuvuduko (m/z n’igihe cyo kubika (RT)) byakozwe hakoreshejwe Compound Discovery 3.1 na TraceFinder 4.0 (Thermo Fisher Scientific). Kugira ngo hakurweho itandukaniro rishoboka hagati y’ibice, buri gice cy’icyitegererezo cy’ikizamini cyagabanijwemo igipimo cy’icyitegererezo cy’ibikoresho by’ibanze bivuye mu itsinda rimwe kugira ngo haboneke ubwinshi bungana. Itandukaniro risanzwe ry’ibipimo by’imbere mbere na nyuma yo gupima rigaragazwa mu mbonerahamwe y’inyongera ya 6. Itandukaniro riri hagati y’amatsinda yombi ryagaragajwe n’igipimo cy’impimbano cyo kuvumbura (FDR<0.05, ikizamini cya Wilcoxon cyashyizweho umukono) n’impinduka (>1.2 cyangwa <0.83). Amakuru ya MS y’ibintu byakuwemo n’amakuru ya MS yahinduwe na serum agaragazwa mu makuru y’inyongera ya 1 n’amakuru y’inyongera ya 2, uko bikurikirana. Incamake y’icyitegererezo yakozwe hashingiwe ku nzego enye zagenwe zo kumenya, harimo metabolites zabonetse, ibintu byavuzwe, ubwoko bw’ibipimo, n’ibintu bitazwi 22. Hashingiwe ku bushakashatsi bwakorewe kuri database muri Compound Discovery 3.1 (mzCloud, HMDB, Chemspider), ibinyabutabire bifite ibipimo byemejwe na MS/MS cyangwa ibisobanuro bihuye neza muri mzCloud (amanota > 85) cyangwa Chemspider byatoranyijwe nk'ibihuza hagati ya metabolome inyuranye. Ibisobanuro by'ingenzi kuri buri kintu biri muri Supplementary Data 3. MetaboAnalyst 5.0 yakoreshejwe mu isesengura ryihariye ry'ubwinshi bwa metabolite busanzwe. MetaboAnalyst 5.0 yanasuzumye isesengura rya KEGG rishingiye kuri metabolite zitandukanye cyane. Isesengura ry'ibice by'ingenzi (PCA) na partial least squares discriminant analysis (PLS-DA) byasesenguwe hakoreshejwe porogaramu ya ropls (v.1.26.4) hamwe na stack normalization na autoscaling. Modeli nziza ya metabolite biomarker yo guhanura kanseri y'udukoko yakozwe hakoreshejwe binary logistic regression hamwe na least absolute shrinkage and selection operator (LASSO, R package v.4.1-3). Imikorere y'icyitegererezo cy'itandukaniro mu matsinda yo gupima no kwemeza yagaragajwe no gupima AUC hashingiwe ku isesengura rya ROC hakurikijwe paki ya pROC (v.1.18.0.). Igabanuka ry'amahirwe meza ryabonetse hashingiwe ku gipimo ntarengwa cya Youden cy'icyitegererezo (ubwitonzi + umwihariko - 1). Ingero zifite agaciro kari munsi cyangwa kanini y'umubare w'ibipimo zizagenwa nk'udusimba tudakomeye na adenocarcinoma y'ibihaha, uko bikurikirana.
Uturemangingo twa A549 (#CCL-185, American Type Culture Collection) twahinzwe mu buryo bwa F-12K burimo 10% FBS. Uturemangingo twa RNA (shRNA) dufata SLC7A5 n'agakoresho ko kugenzura katagamije (NC) byashyizwe muri vector ya lentiviral pLKO.1-puro. Uturemangingo twa shSLC7A5 two kurwanya ihungabana ni ibi bikurikira: Sh1 (5′-GGAGAAACCTGAACAGTT-3′), Sh2 (5′-GCCGTGGACTTCGGGAACTAT-3′). Uturemangingo twa SLC7A5 (#5347) na tubulin (#2148) twaguzwe muri Cell Signaling Technology. Uturemangingo twa SLC7A5 na tubulin twakoreshejwe ku gipimo cya 1:1000 mu isesengura rya Western blot.
Ikizamini cya Seahorse XF Glycolytic Stress gipima urugero rwa aside iri hanze y'umubiri (ECAR). Mu isuzuma, isukari, oligomycin A, na 2-DG byatanzwe bikurikiranye kugira ngo hamenyekane ubushobozi bwa glycolytic bw'uturemangingo nk'uko byapimwe na ECAR.
Uturemangingo twa A549 twashyizwemo ibikoresho byo kugenzura bidashingiye ku ntego (NC) na shSLC7A5 (Sh1, Sh2) byashyizwe mu masahani ya cm 10 y'umurambararo. Ibikoresho bya metabolite byakuwemo ml 1 ya methanol y'amazi ikonje cyane ya 80%. Uturemangingo twa methanol twakuwemo, dushyirwa mu muyoboro mushya, hanyuma dushyirwa kuri centrifuge kuri 15.000 × g mu gihe cy'iminota 15 kuri 4°C. Gukusanya 800 µl ya supernatant hanyuma biyumutse hakoreshejwe icyuma gipima ubushyuhe. Uduce twa metabolite twumye twasesenguwe kugira ngo harebwe urugero rwa tryptophan hakoreshejwe LC-MS/MS nk'uko byasobanuwe haruguru. Urugero rwa NAD (H) mu turemangingo twa A549 (NC na shSLC7A5) rwapimwe hakoreshejwe uburyo bwa NAD+/NADH colorimetric kit (#K337, BioVision) hakurikijwe amabwiriza y'uwakoze. Urugero rwa poroteyine rwapimwe kuri buri gipimo kugira ngo ingano ya metabolite irusheho kumera neza.
Nta buryo bw'ibarurishamibare bwakoreshejwe kugira ngo hamenyekane ingano y'icyitegererezo. Inyigo za metabolomike zabanje zari zigamije kuvumbura biomarker15,18 zafashwe nk'ibipimo ngenderwaho byo kumenya ingano, kandi ugereranije n'izi raporo, icyitegererezo cyacu cyari gihagije. Nta ngero zakuwe mu itsinda ry'ubushakashatsi. Ingero za serum zashyizwe mu itsinda ry'ubushakashatsi ku buryo butunguranye (abantu 306, 74.6%) n'itsinda ry'igenzura ry'imbere (abantu 104, 25.4%) ku bushakashatsi bwa metabolomike butari bugamije. Twatoranyije kandi abantu 70 muri buri tsinda mu buryo butunguranye mu itsinda ry'ubushakashatsi ku bushakashatsi bwa metabolomike bugamije. Abashakashatsi bahumishijwe amaso ku ishyirwa mu matsinda mu gihe cyo gukusanya no gusesengura amakuru ya LC-MS. Isesengura ry'ibarurishamibare ry'amakuru ya metabolomike n'igerageza ry'uturemangingo risobanurwa mu bice by'ibisubizo, Imiterere y'Ishusho, n'Uburyo. Isuzuma rya tryptophan y'uturemangingo, NADT, n'ibikorwa bya glycolytic ryakozwe inshuro eshatu ku giti cyaryo, hagatangwa ibisubizo bimwe.
Kugira ngo umenye byinshi ku bijyanye n'igishushanyo mbonera cy'ubushakashatsi, reba Incamake ya Raporo y'Ubushakashatsi bw'Ibintu Bisanzwe ijyanye n'iyi nkuru.
Amakuru ya MS y’ibintu byakuwemo hamwe n’amakuru ya MS asanzwe ya serumu y’inyongera agaragara muri Supplementary Data 1 na Supplementary Data 2, uko bikurikirana. Ibisobanuro by’ingenzi ku bintu bitandukanye bigaragazwa muri Supplementary Data 3. Amakuru ya LUAD TCGA ashobora gukurwa kuri https://portal.gdc.cancer.gov/. Amakuru yinjiye yo gushushanya imbonerahamwe aboneka mu makuru y’inyongera. Amakuru y’inyongera atangwa kuri iyi ngingo.
Itsinda ry'Igihugu ry'Ubushakashatsi ku Bihaha, n'ibindi. Kugabanya impfu ziterwa na kanseri y'ibihaha hakoreshejwe uburyo bwo gupima ku gipimo gito. Ubwongereza bw'Amajyaruguru. J. Med. 365, 395–409 (2011).
Kramer, BS, Berg, KD, Aberle, DR na Prophet, Gupima kanseri y'ibihaha hakoreshejwe CT yo mu bwoko bwa helical CT: ibyavuye mu bushakashatsi bw'igihugu bwo gusuzuma ibihaha (NLST). J. Med. Screen 18, 109–111 (2011).
De Koning, HJ, n'abandi. Kugabanya impfu ziterwa na kanseri y'ibihaha hakoreshejwe uburyo bwa CT mu igerageza ryakozwe ku buryo butunguranye. Amajyaruguru y'Ubwongereza. J. Med. 382, 503–513 (2020).
Igihe cyo kohereza ubutumwa: 18 Nzeri 2023