Kev kuaj mob sib txawv ntawm cov qog hauv lub ntsws uas tau txheeb xyuas los ntawm kev siv computer tomography (CT) tseem yog ib qho kev sib tw hauv kev kho mob. Ntawm no, peb piav qhia txog lub ntiaj teb metabolilome ntawm 480 cov qauv serum, suav nrog cov neeg noj qab haus huv, cov qog hauv lub ntsws benign, thiab theem I mob qog hauv lub ntsws adenocarcinoma. Cov qog hauv lub ntsws adenocarcinomas muaj cov qauv metabolilomic tshwj xeeb, thaum cov qog benign thiab cov neeg noj qab haus huv muaj kev zoo sib xws hauv cov qauv metabolilomic. Hauv pawg tshawb pom (n = 306), ib pawg ntawm 27 metabolites tau txheeb xyuas kom paub qhov txawv ntawm cov qog benign thiab malignant. AUC ntawm tus qauv discriminant hauv kev lees paub sab hauv (n = 104) thiab kev lees paub sab nraud (n = 111) pawg yog 0.915 thiab 0.945, raws li. Kev tshuaj xyuas txoj kev tau qhia txog kev nce ntxiv ntawm cov metabolites glycolytic cuam tshuam nrog kev txo qis tryptophan hauv cov qog hauv lub ntsws adenocarcinoma serum piv rau cov qog benign thiab cov neeg noj qab haus huv, thiab qhia tias tryptophan uptake txhawb nqa glycolysis hauv cov hlwb mob qog hauv lub ntsws. Peb txoj kev tshawb fawb qhia txog qhov tseem ceeb ntawm cov cim qhia txog cov metabolite biomarkers hauv cov ntshav hauv kev ntsuam xyuas qhov kev pheej hmoo ntawm cov qog hauv lub ntsws uas pom los ntawm CT.
Kev kuaj mob thaum ntxov yog qhov tseem ceeb heev rau kev txhim kho cov neeg mob qog noj ntshav kom muaj sia nyob. Cov txiaj ntsig los ntawm US National Lung Cancer Screening Trial (NLST) thiab European NELSON Study tau qhia tias kev tshuaj ntsuam nrog cov tshuaj suav qis (LDCT) tuaj yeem txo qhov kev tuag ntawm qog noj ntshav hauv cov pab pawg muaj kev pheej hmoo siab1,2,3. Txij li thaum siv LDCT dav dav rau kev tshuaj ntsuam qog noj ntshav hauv lub ntsws, qhov tshwm sim ntawm kev tshawb pom radiographic ntawm cov qog ntshav hauv lub ntsws tsis muaj tsos mob tau txuas ntxiv nce 4. Cov qog ntshav hauv lub ntsws yog txhais tias yog qhov tsis pom tseeb txog li 3 cm hauv txoj kab uas hla 5. Peb ntsib teeb meem hauv kev ntsuam xyuas qhov yuav tshwm sim ntawm kev mob qog noj ntshav thiab kev daws teeb meem nrog cov qog ntshav hauv lub ntsws loj uas pom tsis tseeb ntawm LDCT. Kev txwv ntawm CT tuaj yeem ua rau muaj kev kuaj mob ntau zaus thiab cov txiaj ntsig tsis tseeb, ua rau muaj kev cuam tshuam tsis tsim nyog thiab kev kho mob ntau dhau6. Yog li ntawd, muaj qhov xav tau los tsim cov cim qhia txog kev ntseeg tau thiab muaj txiaj ntsig los txheeb xyuas cov qog ntshav hauv lub ntsws kom raug thaum ntxov thiab sib txawv feem ntau cov qog ntshav zoo thaum pib kuaj pom 7.
Kev tshuaj xyuas cov ntshav (serum, plasma, peripheral ntshav mononuclear cells), suav nrog genomics, proteomics lossis DNA methylation8,9,10, tau ua rau muaj kev txaus siab rau kev tshawb pom cov cim qhia txog kev mob qog noj ntshav hauv lub ntsws. Lub caij no, cov txheej txheem metabolimics ntsuas cov khoom kawg ntawm tes uas cuam tshuam los ntawm kev ua endogenous thiab exogenous thiab yog li ntawd siv los kwv yees qhov pib ntawm kab mob thiab qhov tshwm sim. Kua chromatography-tandem mass spectrometry (LC-MS) yog ib txoj hauv kev siv dav rau kev tshawb fawb metabolomics vim nws qhov kev nkag siab siab thiab ntau yam dynamic, uas tuaj yeem npog cov metabolites nrog cov khoom sib txawv physicochemical11,12,13. Txawm hais tias kev tshuaj xyuas thoob ntiaj teb ntawm plasma / serum tau siv los txheeb xyuas cov cim qhia txog kev mob qog noj ntshav hauv lub ntsws14,15,16,17 thiab kev kho mob zoo,18 serum metabolite classifiers kom paub qhov txawv ntawm cov qog nqaij hlav benign thiab malignant tseem yuav tsum tau kawm ntau. -kev tshawb fawb loj heev.
Adenocarcinoma thiab squamous cell carcinoma yog ob hom tseem ceeb ntawm cov mob qog nqaij hlav tsis yog me me (NSCLC). Ntau yam kev kuaj CT qhia tau tias adenocarcinoma yog hom mob qog nqaij hlav feem ntau1,19,20,21. Hauv kev tshawb fawb no, peb tau siv ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) los ua qhov kev tshuaj xyuas metabolimomics ntawm tag nrho 695 cov qauv serum, suav nrog cov neeg noj qab haus huv, cov qog nqaij hlav benign, thiab CT-nrhiav tau ≤3 cm. Kev tshuaj xyuas rau Theem I mob qog nqaij hlav ntsws. Peb tau txheeb xyuas ib pawg ntawm cov metabolites serum uas sib txawv ntawm cov qog nqaij hlav ntsws los ntawm cov qog nqaij hlav benign thiab cov neeg noj qab haus huv. Kev tshuaj xyuas txoj kev enrichment tau qhia tias tryptophan tsis zoo thiab glucose metabolism yog kev hloov pauv ntau hauv cov qog nqaij hlav ntsws piv rau cov qog nqaij hlav benign thiab cov neeg noj qab haus huv. Thaum kawg, peb tau tsim thiab lees paub qhov kev faib tawm ntawm cov tshuaj lom neeg hauv cov ntshav uas muaj qhov tshwj xeeb thiab rhiab heev kom paub qhov txawv ntawm cov qog nqaij hlav malignant thiab benign uas pom los ntawm LDCT, uas yuav pab tau rau kev kuaj mob sib txawv thaum ntxov thiab kev ntsuas kev pheej hmoo.
Hauv kev tshawb fawb tam sim no, cov qauv ntshav sib phim poj niam txiv neej thiab hnub nyoog tau raug sau rov qab los ntawm 174 tus neeg tswj kev noj qab haus huv, 292 tus neeg mob uas muaj cov qog ntshav hauv lub ntsws tsis muaj mob, thiab 229 tus neeg mob uas muaj theem I mob qog ntshav hauv lub ntsws. Cov yam ntxwv ntawm cov pej xeem ntawm 695 tus neeg tau qhia nyob rau hauv Daim Ntawv Ntxiv 1.
Raws li tau pom hauv Daim Duab 1a, tag nrho muaj 480 cov qauv serum, suav nrog 174 tus neeg noj qab nyob zoo (HC), 170 tus mob qog nqaij hlav tsis muaj qog (BN), thiab 136 tus mob qog nqaij hlav ntsws theem I (LA) tau raug sau ntawm Sun Yat-sen University Cancer Center. Cov neeg tshawb pom rau kev tshawb nrhiav metabolomic profiling siv ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). Raws li tau pom hauv Daim Duab Ntxiv 1, cov metabolites sib txawv ntawm LA thiab HC, LA thiab BN tau raug txheeb xyuas los tsim kom muaj tus qauv kev faib tawm thiab tshawb nrhiav ntxiv txog kev tshuaj xyuas txoj kev sib txawv. 104 cov qauv uas tau sau los ntawm Sun Yat-sen University Cancer Center thiab 111 cov qauv uas tau sau los ntawm ob lub tsev kho mob tau raug kuaj xyuas sab hauv thiab sab nraud, raws li.
a Cov pej xeem kawm hauv pawg neeg tshawb pom uas tau dhau los ntawm kev tshuaj xyuas thoob ntiaj teb serum metabolomics siv ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). b Ib feem tsawg kawg nkaus squares discriminant analysis (PLS-DA) ntawm tag nrho cov metabolome ntawm 480 serum qauv los ntawm pawg neeg tshawb fawb, suav nrog cov neeg tswj kev noj qab haus huv (HC, n = 174), benign nodules (BN, n = 170), thiab theem I lub ntsws adenocarcinoma (Los Angeles, n = 136). +ESI, positive electrospray ionization mode, -ESI, negative electrospray ionization mode. c-e Metabolites nrog cov khoom sib txawv ntau hauv ob pawg muab (two-tailed Wilcoxon signed rank test, false discovery rate adjusted p value, FDR <0.05) tau qhia ua liab (fold change > 1.2) thiab xiav (fold change < 0.83). ) tau qhia rau ntawm daim duab roob hluav taws. f Daim ntawv qhia kub ntawm pawg hierarchical qhia txog qhov sib txawv tseem ceeb ntawm tus lej ntawm cov metabolites uas tau sau tseg ntawm LA thiab BN. Cov ntaub ntawv tau muab los ua cov ntaub ntawv qhov chaw.
Tag nrho cov serum metabolome ntawm 174 HC, 170 BN thiab 136 LA hauv pawg tshawb pom tau raug tshuaj xyuas siv UPLC-HRMS kev tshuaj xyuas. Peb thawj zaug qhia tias cov qauv tswj xyuas zoo (QC) sib sau ua ke nruj nreem ntawm qhov chaw ntawm tus qauv tshuaj xyuas cov khoom tseem ceeb tsis muaj tus saib xyuas (PCA), lees paub qhov ruaj khov ntawm kev ua tau zoo ntawm kev tshawb fawb tam sim no (Daim Duab Ntxiv 2).
Raws li tau pom hauv qhov kev tshuaj xyuas partial least squares-discriminant (PLS-DA) hauv Daim Duab 1 b, peb pom tias muaj qhov sib txawv meej ntawm LA thiab BN, LA thiab HC hauv cov hom electrospray ionization zoo (+ESI) thiab tsis zoo (-ESI). Txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb pom ntawm BN thiab HC hauv +ESI thiab -ESI tej yam kev mob.
Peb pom 382 yam ntxwv sib txawv ntawm LA thiab HC, 231 yam ntxwv sib txawv ntawm LA thiab BN, thiab 95 yam ntxwv sib txawv ntawm BN thiab HC (Wilcoxon kos npe qib xeem, FDR <0.05 thiab ntau qhov kev hloov pauv >1.2 lossis <0.83) (Daim Duab .1c-e). . Cov ncov tau ntxiv sau tseg (Cov Ntaub Ntawv Ntxiv 3) tawm tsam lub ntaub ntawv (mzCloud/HMDB/Chemspider tsev qiv ntawv) los ntawm m/z tus nqi, lub sijhawm khaws cia thiab kev tshawb nrhiav pawg loj spectrum (cov ntsiab lus piav qhia hauv ntu Txoj Kev) 22. Thaum kawg, 33 thiab 38 cov metabolites sau tseg nrog qhov sib txawv tseem ceeb hauv kev nplua mias tau txheeb xyuas rau LA piv rau BN (Daim Duab 1f thiab Cov Lus Qhia Ntxiv 2) thiab LA piv rau HC (Daim Duab Ntxiv 3 thiab Cov Lus Qhia Ntxiv 2), raws li. Qhov sib piv, tsuas yog 3 cov metabolites nrog qhov sib txawv tseem ceeb hauv kev nplua mias tau txheeb xyuas hauv BN thiab HC (Cov Lus Qhia Ntxiv 2), sib xws nrog kev sib tshooj ntawm BN thiab HC hauv PLS-DA. Cov metabolites sib txawv no npog ntau yam biochemicals (Daim Duab Ntxiv 4). Thaum muab ua ke, cov txiaj ntsig no qhia txog kev hloov pauv tseem ceeb hauv cov serum metabolome uas qhia txog kev hloov pauv malignant ntawm mob qog noj ntshav thaum ntxov piv rau cov qog nqaij hlav benign lossis cov neeg noj qab haus huv. Lub caij no, qhov zoo sib xws ntawm cov serum metabolome ntawm BN thiab HC qhia tias cov qog nqaij hlav benign yuav muaj ntau yam ntxwv ntawm cov tib neeg noj qab haus huv. Muab hais tias epidermal growth factor receptor (EGFR) gene mutations yog qhov tshwm sim hauv lub ntsws adenocarcinoma subtype 23, peb nrhiav kev txiav txim siab qhov cuam tshuam ntawm cov tsav tsheb mutations rau ntawm serum metabolome. Tom qab ntawd peb tau tshuaj xyuas tag nrho cov ntaub ntawv metabolilomic ntawm 72 tus neeg mob nrog EGFR xwm txheej hauv pawg ntsws adenocarcinoma. Qhov nthuav yog, peb pom cov ntaub ntawv sib piv ntawm EGFR mutant cov neeg mob (n = 41) thiab EGFR wild-type cov neeg mob (n = 31) hauv PCA kev tshuaj xyuas (Daim Duab Ntxiv 5a). Txawm li cas los xij, peb tau txheeb xyuas 7 lub metabolites uas lawv qhov ntau tau hloov pauv ntau hauv cov neeg mob uas muaj EGFR mutation piv rau cov neeg mob uas muaj hom EGFR qus (t xeem, p <0.05 thiab hloov pauv ntau > 1.2 lossis <0.83) (Daim Duab Ntxiv 5b). Feem ntau ntawm cov metabolites no (5 ntawm 7) yog acylcarnitines, uas ua lub luag haujlwm tseem ceeb hauv cov txheej txheem oxidation fatty acid.
Raws li tau piav qhia hauv cov txheej txheem ua haujlwm uas tau qhia hauv Daim Duab 2 a, cov cim qhia txog kev faib tawm ntawm cov nodule tau txais los ntawm kev siv cov neeg ua haujlwm tsawg kawg nkaus thiab kev xaiv raws li 33 cov metabolites sib txawv uas tau txheeb xyuas hauv LA (n = 136) thiab BN (n = 170). Kev sib xyaw ua ke zoo tshaj plaws ntawm cov hloov pauv (LASSO) - qauv binary logistic regression. Kev lees paub hla kaum npaug tau siv los sim qhov kev ntseeg siab ntawm tus qauv. Kev xaiv hloov pauv thiab kev ua kom tsis raug cai tau kho los ntawm kev nplua rau qhov zoo li qub nrog parameter λ24. Kev tshuaj xyuas thoob ntiaj teb metabolomics tau ua tiav ntxiv ywj pheej hauv kev lees paub sab hauv (n = 104) thiab kev lees paub sab nraud (n = 111) pawg los sim kev ua tau zoo ntawm kev faib tawm ntawm tus qauv discriminant. Yog li ntawd, 27 metabolites hauv cov txheej txheem tshawb pom tau txheeb xyuas tias yog tus qauv discriminant zoo tshaj plaws nrog tus nqi AUC loj tshaj plaws (Daim Duab 2b), ntawm cov uas 9 tau ua haujlwm ntau ntxiv thiab 18 txo qis hauv LA piv rau BN (Daim Duab 2c).
Cov txheej txheem ua haujlwm rau kev tsim cov kab mob ntsws pulmonary nodule classifier, suav nrog kev xaiv cov vaj huam sib luag zoo tshaj plaws ntawm cov metabolites hauv cov txheej txheem tshawb pom siv cov qauv binary logistic regression ntawm kaum-zaug hla-validation thiab kev soj ntsuam kev ua tau zoo hauv cov txheej txheem lees paub sab hauv thiab sab nraud. b Cov ntaub ntawv hla-validation ntawm LASSO regression qauv rau kev xaiv biomarker metabolic. Cov lej muab saum toj no sawv cev rau tus lej nruab nrab ntawm cov biomarkers xaiv ntawm ib qho λ. Cov kab liab dotted sawv cev rau tus nqi AUC nruab nrab ntawm lambda sib xws. Cov kab yuam kev grey sawv cev rau qhov tsawg kawg nkaus thiab siab tshaj plaws AUC tus nqi. Cov kab dotted qhia txog tus qauv zoo tshaj plaws nrog 27 biomarkers xaiv. AUC, thaj chaw nyob rau hauv tus txais kev ua haujlwm tus cwj pwm (ROC) nkhaus. c Quav kev hloov pauv ntawm 27 cov metabolites xaiv hauv pawg LA piv nrog pawg BN hauv pawg tshawb pom. Kab liab - kev ua haujlwm. Kab xiav yog qhov poob qis. d-f Cov kab Receiver tus cwj pwm ua haujlwm (ROC) qhia lub zog ntawm tus qauv discriminant raws li 27 metabolite ua ke hauv cov txheej txheem tshawb pom, sab hauv, thiab sab nraud. Cov ntaub ntawv qhov chaw yog muab rau hauv daim ntawv ntawm cov ntaub ntawv qhov chaw.
Ib qho qauv kwv yees tau tsim los ntawm cov coefficients regression hnyav ntawm 27 metabolites no (Cov Lus Qhia Ntxiv 3). ROC kev tshuaj xyuas raws li 27 metabolites no tau muab thaj chaw hauv qab qhov nkhaus (AUC) tus nqi ntawm 0.933, kev nkag siab ntawm pawg tshawb pom yog 0.868, thiab qhov tshwj xeeb yog 0.859 (Daim Duab 2d). Lub caij no, ntawm 38 cov metabolites sib txawv ntawm LA thiab HC, ib pawg ntawm 16 metabolites tau ua tiav AUC ntawm 0.902 nrog qhov rhiab heev ntawm 0.801 thiab qhov tshwj xeeb ntawm 0.856 hauv kev cais LA los ntawm HC (Daim Duab Ntxiv 6a-c). AUC tus nqi raws li cov kev hloov pauv sib txawv rau cov metabolites sib txawv kuj tau piv. Peb pom tias tus qauv kev faib tawm ua tau zoo tshaj plaws hauv kev cais ntawm LA thiab BN (HC) thaum qib hloov pauv tau teeb tsa rau 1.2 piv rau 1.5 lossis 2.0 (Daim Duab Ntxiv 7a, b). Tus qauv kev faib tawm, raws li 27 pawg metabolite, tau raug lees paub ntxiv hauv cov pawg sab hauv thiab sab nraud. AUC yog 0.915 (kev rhiab heev 0.867, qhov tshwj xeeb 0.811) rau kev lees paub sab hauv thiab 0.945 (kev rhiab heev 0.810, qhov tshwj xeeb 0.979) rau kev lees paub sab nraud (Daim duab 2e, f). Txhawm rau ntsuas qhov ua tau zoo ntawm kev sib koom ua ke, 40 cov qauv los ntawm pawg sab nraud tau raug tshuaj xyuas hauv chav kuaj sab nraud raws li tau piav qhia hauv ntu Txoj Kev. Qhov tseeb ntawm kev faib tawm tau ua tiav AUC ntawm 0.925 (Daim Duab Ntxiv 8). Vim tias mob ntsws squamous cell carcinoma (LUSC) yog hom mob ntsws thib ob uas tsis yog mob me me (NSCLC) tom qab mob ntsws adenocarcinoma (LUAD), peb kuj tau sim qhov muaj peev xwm siv tau ntawm cov qauv metabolic. BN thiab 16 rooj plaub ntawm LUSC. Tus nqi AUC ntawm kev sib cais ntawm LUSC thiab BN yog 0.776 (Daim Duab Ntxiv 9), qhia txog kev muaj peev xwm tsis zoo piv rau kev sib cais ntawm LUAD thiab BN.
Cov kev tshawb fawb tau qhia tias qhov loj ntawm cov nodules ntawm CT duab muaj feem cuam tshuam zoo nrog qhov yuav muaj mob qog noj ntshav thiab tseem yog qhov tseem ceeb ntawm kev kho mob nodule25,26,27. Kev tshuaj xyuas cov ntaub ntawv los ntawm pawg neeg loj ntawm NELSON kev tshuaj ntsuam xyuas qhia tau hais tias qhov kev pheej hmoo ntawm mob qog noj ntshav hauv cov neeg uas muaj cov nodes <5 hli zoo ib yam li cov neeg uas tsis muaj nodes 28. Yog li ntawd, qhov tsawg kawg nkaus uas xav tau kev saib xyuas CT tas li yog 5 hli, raws li British Thoracic Society (BTS) pom zoo, thiab 6 hli, raws li Fleischner Society 29 pom zoo. Txawm li cas los xij, cov nodules loj dua 6 hli thiab tsis muaj cov yam ntxwv zoo, hu ua indeterminate pulmonary nodules (IPN), tseem yog qhov nyuaj loj hauv kev ntsuam xyuas thiab kev tswj hwm hauv kev kho mob30,31. Tom ntej no peb tau tshuaj xyuas seb qhov loj ntawm nodule puas cuam tshuam rau cov cim metabolomic siv cov qauv sib sau ua ke los ntawm kev tshawb pom thiab kev lees paub sab hauv. Tsom mus rau 27 cov cim biomarkers, peb thawj zaug piv rau PCA profiles ntawm HC thiab BN sub-6 hli metabolomes. Peb pom tias feem ntau ntawm cov ntaub ntawv rau HC thiab BN sib tshooj, qhia tau tias cov theem metabolite hauv cov ntshav zoo sib xws hauv ob pawg (Daim Duab 3a). Cov ntawv qhia txog cov yam ntxwv ntawm ntau qhov loj sib txawv tseem khaws cia hauv BN thiab LA (Daim Duab 3b, c), thaum pom kev sib cais ntawm cov qog nqaij hlav phem thiab cov qog nqaij hlav tsis muaj zog hauv 6–20 hli (Daim Duab 3d). Pawg neeg no muaj AUC ntawm 0.927, qhov tshwj xeeb ntawm 0.868, thiab qhov rhiab heev ntawm 0.820 rau kev kwv yees qhov mob qog nqaij hlav ntawm cov qog nqaij hlav ntsuas 6 txog 20 hli (Daim Duab 3e, f). Peb cov txiaj ntsig qhia tau tias tus neeg faib tawm tuaj yeem ntes cov kev hloov pauv metabolic uas tshwm sim los ntawm kev hloov pauv malignant thaum ntxov, tsis hais qhov loj ntawm cov qog nqaij hlav.
kev sib piv ntawm PCA profiles ntawm cov pab pawg tshwj xeeb raws li kev faib tawm metabolic ntawm 27 metabolites. CC thiab BN <6 hli. b BN <6 hli vs BN 6–20 hli. hauv LA 6–20 hli piv rau LA 20–30 hli. g BN 6–20 hli thiab LA 6–20 hli. GC, n = 174; BN <6 hli, n = 153; BN 6–20 hli, n = 91; LA 6–20 hli, n = 89; LA 20–30 hli, n = 77. e Tus txais kev ua haujlwm yam ntxwv (ROC) nkhaus qhia txog kev ua haujlwm qauv discriminant rau cov nodules 6–20 hli. f Cov nqi probability tau suav raws li tus qauv logistic regression rau cov nodules ntsuas 6–20 hli. Cov kab grey dotted sawv cev rau qhov zoo tshaj plaws cutoff nqi (0.455). Cov lej saum toj no sawv cev rau feem pua ntawm cov xwm txheej kwv yees rau Los Angeles. Siv qhov kev xeem t uas muaj ob sab. PCA, kev tshuaj xyuas cov khoom tseem ceeb. Thaj chaw AUC nyob hauv qab kab nkhaus. Cov ntaub ntawv qhov chaw tau muab los ua cov ntaub ntawv qhov chaw.
Plaub tus qauv (hnub nyoog 44–61 xyoo) nrog cov qhov loj me zoo sib xws ntawm lub ntsws (7–9 hli) tau raug xaiv ntxiv los qhia txog kev ua tau zoo ntawm tus qauv kwv yees mob qog noj ntshav (Daim Duab 4a, b). Thaum kuaj thawj zaug, Case 1 tau nthuav tawm ua ib lub qhov khov kho nrog calcification, ib qho tshwj xeeb cuam tshuam nrog kev mob qog noj ntshav, thaum Case 2 tau nthuav tawm ua ib lub qhov khov kho tsis paub meej uas tsis muaj cov yam ntxwv zoo. Peb qhov kev soj ntsuam CT tom qab tau qhia tias cov rooj plaub no tseem ruaj khov dhau 4-xyoo thiab yog li ntawd tau suav tias yog cov qhov khov kho (Daim Duab 4a). Piv nrog kev soj ntsuam kev kho mob ntawm cov CT scans, kev tshuaj xyuas metabolite ib zaug nrog tus qauv classifier tam sim no tau txheeb xyuas cov qhov khov kho no sai thiab raug raws li cov kev txwv probabilistic (Rooj 1). Daim Duab 4b hauv rooj plaub 3 qhia txog lub qhov khov kho nrog cov cim ntawm pleural retraction, uas feem ntau cuam tshuam nrog mob qog noj ntshav32. Case 4 tau nthuav tawm ua ib lub qhov khov kho tsis paub meej uas tsis muaj pov thawj ntawm qhov ua rau mob qog noj ntshav. Tag nrho cov rooj plaub no tau kwv yees tias yog mob qog noj ntshav raws li tus qauv faib tawm (Rooj 1). Kev ntsuam xyuas ntawm lub ntsws adenocarcinoma tau pom tseeb los ntawm kev kuaj mob histopathological tom qab phais mob ntsws (Daim duab 4b). Rau cov txheej txheem lees paub sab nraud, tus neeg faib tawm metabolic tau kwv yees qhov tseeb ob rooj plaub ntawm cov qog ntsws tsis paub meej loj dua 6 hli (Daim duab ntxiv 10).
Cov duab CT ntawm lub qhov rais axial ntawm lub ntsws ntawm ob rooj plaub ntawm cov nodules benign. Hauv rooj plaub 1, CT scan tom qab 4 xyoos qhia tau tias muaj ib lub nodule khov kho ntsuas 7 hli nrog calcification hauv sab xis qis. Hauv rooj plaub 2, CT scan tom qab 5 xyoos qhia tau tias muaj ib lub nodule khov kho, ib nrab khov kho nrog lub cheeb ntawm 7 hli hauv sab xis sab saud. b Qhov rai axial CT cov duab ntawm lub ntsws thiab kev tshawb fawb txog kab mob ntawm ob rooj plaub ntawm theem I adenocarcinoma ua ntej phais lub ntsws. Rooj plaub 3 qhia tau tias muaj ib lub nodule nrog lub cheeb ntawm 8 hli hauv sab xis sab saud nrog pleural retraction. Rooj plaub 4 qhia tau tias muaj ib lub nodule av-iav khov kho ib nrab ntsuas 9 hli hauv sab laug sab saud. Hematoxylin thiab eosin (H&E) staining ntawm cov ntaub so ntswg ntsws uas raug phais (scale bar = 50 μm) qhia txog tus qauv kev loj hlob acinar ntawm lub ntsws adenocarcinoma. Cov xub qhia txog cov nodules pom ntawm CT duab. H&E cov duab yog cov duab sawv cev ntawm ntau (> 3) microscopic teb kuaj xyuas los ntawm tus kws kho mob.
Thaum muab peb cov txiaj ntsig los ua ke qhia tau tias muaj txiaj ntsig zoo ntawm cov cim qhia txog cov metabolite biomarkers hauv kev kuaj mob sib txawv ntawm cov qog hauv lub ntsws, uas yuav ua rau muaj kev cov nyom thaum soj ntsuam CT kev kuaj mob.
Raws li qhov kev tshuaj xyuas metabolite sib txawv uas tau lees paub, peb nrhiav kev txheeb xyuas cov kab mob sib raug zoo ntawm cov kev hloov pauv loj hauv metabolic. KEGG txoj kev tshuaj xyuas kev txhim kho los ntawm MetaboAnalyst tau txheeb xyuas 6 txoj kev hloov pauv tseem ceeb ntawm ob pawg muab (LA vs. HC thiab LA vs. BN, kho p ≤ 0.001, nyhuv > 0.01). Cov kev hloov pauv no tau piav qhia los ntawm kev cuam tshuam hauv pyruvate metabolism, tryptophan metabolism, niacin thiab nicotinamide metabolism, glycolysis, TCA lub voj voog, thiab purine metabolism (Daim duab 5a). Tom qab ntawd peb ua tiav cov metabolomics tsom mus rau kev txheeb xyuas cov kev hloov pauv loj siv kev ntsuas qhov tseeb. Kev txiav txim siab ntawm cov metabolites feem ntau hauv cov kev hloov pauv feem ntau los ntawm triple quadrupole mass spectrometry (QQQ) siv cov qauv metabolite tseeb. Cov yam ntxwv ntawm cov neeg kawm txog kev tshawb fawb txog metabolomics tau suav nrog hauv Daim Ntawv Qhia Ntxiv 4. Sib xws nrog peb cov txiaj ntsig thoob ntiaj teb ntawm metabolomics, kev tshuaj xyuas ntau yam tau lees paub tias hypoxanthine thiab xanthine, pyruvate, thiab lactate tau nce ntxiv hauv LA piv rau BN thiab HC (Daim Duab 5b, c, p <0.05). Txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb hauv cov metabolites no tau pom ntawm BN thiab HC.
KEGG txoj kev tshuaj xyuas kev txhim kho ntawm cov metabolites sib txawv hauv pawg LA piv rau pawg BN thiab HC. Ib qho kev sim Globaltest ob-tailed tau siv, thiab p tus nqi tau kho siv txoj kev Holm-Bonferroni (kho p ≤ 0.001 thiab qhov loj me ntawm cov nyhuv> 0.01). b-d Cov phiaj xwm Violin qhia txog hypoxanthine, xanthine, lactate, pyruvate, thiab tryptophan theem hauv serum HC, BN, thiab LA txiav txim siab los ntawm LC-MS / MS (n = 70 ib pawg). Cov kab dawb thiab dub dotted qhia txog qhov nruab nrab thiab quartile, feem. e Cov phiaj xwm Violin qhia txog kev ua haujlwm Log2TPM (cov ntawv sau ib lab) mRNA kev qhia ntawm SLC7A5 thiab QPRT hauv lub ntsws adenocarcinoma (n = 513) piv rau cov ntaub so ntswg ib txwm muaj (n = 59) hauv cov ntaub ntawv LUAD-TCGA. Lub thawv dawb sawv cev rau qhov sib txawv ntawm interquartile, kab dub kab rov tav hauv nruab nrab sawv cev rau qhov nruab nrab, thiab kab dub ntsug txuas ntxiv los ntawm lub thawv sawv cev rau 95% kev ntseeg siab (CI). f Pearson correlation plot ntawm SLC7A5 thiab GAPDH kev qhia tawm hauv lub ntsws adenocarcinoma (n = 513) thiab cov ntaub so ntswg ib txwm muaj (n = 59) hauv TCGA dataset. Thaj chaw grey sawv cev rau 95% CI. r, Pearson correlation coefficient. g Cov theem tryptophan cellular uas tau kho kom zoo hauv A549 hlwb transfected nrog kev tswj hwm shRNA tsis tshwj xeeb (NC) thiab shSLC7A5 (Sh1, Sh2) txiav txim siab los ntawm LC-MS/MS. Kev tshuaj xyuas txog tsib qhov qauv ywj pheej hauv txhua pawg tau nthuav tawm. h Cov theem cellular ntawm NADt (tag nrho NAD, suav nrog NAD+ thiab NADH) hauv A549 hlwb (NC) thiab SLC7A5 knockdown A549 hlwb (Sh1, Sh2). Kev tshuaj xyuas txog peb qhov qauv ywj pheej hauv txhua pawg tau nthuav tawm. i Kev ua haujlwm glycolytic ntawm A549 hlwb ua ntej thiab tom qab SLC7A5 knockdown tau ntsuas los ntawm extracellular acidification rate (ECAR) (n = 4 biologically ywj pheej kuaj ib pawg). 2-DG,2-deoxy-D-glucose. Two-tailed Student's t test tau siv rau hauv (b-h). Hauv (g-i), cov kab yuam kev sawv cev rau qhov nruab nrab ± SD, txhua qhov kev sim tau ua peb zaug ywj pheej thiab cov txiaj ntsig zoo sib xws. Cov ntaub ntawv qhov chaw tau muab rau hauv daim ntawv ntawm cov ntaub ntawv qhov chaw.
Xav txog qhov cuam tshuam tseem ceeb ntawm kev hloov pauv tryptophan metabolism hauv pawg LA, peb kuj tau soj ntsuam cov serum tryptophan hauv pawg HC, BN, thiab LA siv QQQ. Peb pom tias serum tryptophan tau txo qis hauv LA piv nrog HC lossis BN (p < 0.001, Daim Duab 5d), uas yog sib xws nrog cov kev tshawb pom yav dhau los tias cov tryptophan circulating qis dua hauv cov neeg mob uas muaj mob qog noj ntshav ntsws dua li cov neeg noj qab haus huv los ntawm pawg tswj hwm33,34,35. Lwm txoj kev tshawb fawb siv PET / CT tracer 11C-methyl-L-tryptophan pom tias lub sijhawm khaws cia tryptophan teeb liab hauv cov nqaij mob qog noj ntshav ntsws tau nce ntxiv ntau piv rau cov qhov txhab benign lossis cov nqaij ib txwm36. Peb xav tias qhov txo qis hauv tryptophan hauv LA serum yuav qhia txog kev nqus tryptophan los ntawm cov hlwb qog noj ntshav ntsws.
Nws kuj paub tias qhov khoom kawg ntawm txoj kev kynurenine ntawm tryptophan catabolism yog NAD + 37,38, uas yog ib qho tseem ceeb rau cov tshuaj tiv thaiv ntawm glyceraldehyde-3-phosphate nrog 1,3-bisphosphoglycerate hauv glycolysis39. Thaum cov kev tshawb fawb yav dhau los tau tsom mus rau lub luag haujlwm ntawm tryptophan catabolism hauv kev tswj hwm kev tiv thaiv kab mob, peb nrhiav kev piav qhia txog kev sib cuam tshuam ntawm tryptophan dysregulation thiab glycolytic txoj kev pom hauv kev tshawb fawb tam sim no. Solute transporter tsev neeg 7 tus tswv cuab 5 (SLC7A5) paub tias yog tryptophan transporter43,44,45. Quinolinic acid phosphoribosyltransferase (QPRT) yog ib qho enzyme nyob hauv qab ntawm txoj kev kynurenine uas hloov quinolinic acid rau NAMN46. Kev tshuaj xyuas ntawm LUAD TCGA dataset tau qhia tias ob qho tib si SLC7A5 thiab QPRT tau nce siab hauv cov ntaub so ntswg qog piv rau cov ntaub so ntswg ib txwm muaj (Daim duab 5e). Qhov kev nce no tau pom nyob rau theem I thiab II nrog rau theem III thiab IV ntawm lub ntsws adenocarcinoma (Daim Duab Ntxiv 11), qhia txog kev cuam tshuam thaum ntxov hauv tryptophan metabolism cuam tshuam nrog tumorigenesis.
Tsis tas li ntawd xwb, cov ntaub ntawv LUAD-TCGA tau qhia txog kev sib raug zoo ntawm SLC7A5 thiab GAPDH mRNA kev qhia tawm hauv cov qauv neeg mob qog noj ntshav (r = 0.45, p = 1.55E-26, Daim Duab 5f). Qhov sib txawv, tsis muaj kev sib raug zoo tseem ceeb ntawm cov cim qhia gene hauv cov ntaub so ntswg ib txwm muaj (r = 0.25, p = 0.06, Daim Duab 5f). Kev rhuav tshem ntawm SLC7A5 (Daim Duab Ntxiv 12) hauv A549 hlwb txo qis cov theem tryptophan thiab NAD (H) ntawm tes (Daim Duab 5g, h), ua rau muaj kev ua haujlwm glycolytic tsawg dua raws li ntsuas los ntawm extracellular acidification rate (ECAR) (Daim Duab 1). 5i). Yog li, raws li kev hloov pauv metabolic hauv serum thiab kev kuaj pom hauv vitro, peb xav tias tryptophan metabolism yuav tsim NAD + los ntawm txoj kev kynurenine thiab ua lub luag haujlwm tseem ceeb hauv kev txhawb nqa glycolysis hauv mob qog noj ntshav.
Cov kev tshawb fawb tau qhia tias muaj ntau cov qog nqaij hlav hauv lub ntsws uas tsis paub meej uas LDCT pom tau tias yuav ua rau muaj kev kuaj ntxiv xws li PET-CT, kev kuaj lub ntsws, thiab kev kho mob ntau dhau vim muaj kev kuaj mob tsis tseeb.31 Raws li pom hauv Daim Duab 6, peb txoj kev tshawb fawb tau txheeb xyuas cov metabolites hauv cov ntshav uas muaj txiaj ntsig zoo rau kev kuaj mob uas yuav txhim kho kev faib cov kev pheej hmoo thiab kev tswj hwm tom qab ntawm cov qog nqaij hlav hauv lub ntsws uas CT pom tau.
Cov qog hauv lub ntsws raug soj ntsuam siv cov tshuaj suav hauv computer tsawg (LDCT) nrog cov yam ntxwv duab qhia txog cov ua rau mob qog lossis mob qog. Qhov tshwm sim tsis paub meej ntawm cov qog tuaj yeem ua rau muaj kev mus ntsib kws kho mob ntau zaus, kev cuam tshuam tsis tsim nyog, thiab kev kho mob ntau dhau. Kev suav nrog cov tshuaj cais tawm hauv cov ntshav nrog tus nqi kuaj mob yuav txhim kho kev ntsuam xyuas kev pheej hmoo thiab kev tswj hwm tom qab ntawm cov qog hauv lub ntsws. PET positron emission tomography.
Cov ntaub ntawv los ntawm kev tshawb fawb NLST hauv Tebchaws Meskas thiab kev tshawb fawb European NELSON qhia tias kev tshuaj xyuas cov pab pawg neeg muaj kev pheej hmoo siab nrog kev siv tshuaj tsawg tsawg (LDCT) yuav txo tau kev tuag ntawm mob qog noj ntshav ntsws1,3. Txawm li cas los xij, kev ntsuam xyuas kev pheej hmoo thiab kev tswj hwm kev kho mob tom qab ntawm ntau tus kab mob ntsws uas pom los ntawm LDCT tseem yog qhov nyuaj tshaj plaws. Lub hom phiaj tseem ceeb yog los ua kom zoo dua qhov kev faib tawm kom raug ntawm cov txheej txheem LDCT uas twb muaj lawm los ntawm kev koom ua ke cov cim qhia txog kev ntseeg tau.
Qee cov cim qhia txog cov tshuaj molecular, xws li cov metabolites hauv cov ntshav, tau raug txheeb xyuas los ntawm kev sib piv mob qog noj ntshav hauv lub ntsws nrog cov neeg tswj kev noj qab haus huv15,17. Hauv kev tshawb fawb tam sim no, peb tau tsom mus rau kev siv cov tshuaj ntsuam xyuas serum metabolomics kom paub qhov txawv ntawm cov qog nqaij hlav benign thiab malignant uas pom los ntawm LDCT. Peb piv rau cov tshuaj ntsuam xyuas serum thoob ntiaj teb ntawm kev tswj kev noj qab haus huv (HC), cov qog nqaij hlav benign (BN), thiab theem I mob qog nqaij hlav adenocarcinoma (LA) siv kev tshuaj ntsuam xyuas UPLC-HRMS. Peb pom tias HC thiab BN muaj cov qauv metabolic zoo sib xws, thaum LA tau qhia txog kev hloov pauv tseem ceeb piv rau HC thiab BN. Peb tau txheeb xyuas ob pawg ntawm cov metabolites hauv serum uas sib txawv LA ntawm HC thiab BN.
Cov txheej txheem txheeb xyuas tam sim no raws li LDCT rau cov qog nqaij hlav benign thiab malignant yog raws li qhov loj me, qhov ceev, morphology thiab kev loj hlob ntawm cov qog nqaij hlav dhau sijhawm30. Cov kev tshawb fawb yav dhau los tau qhia tias qhov loj me ntawm cov qog nqaij hlav muaj feem cuam tshuam nrog qhov yuav muaj mob qog nqaij hlav ntsws. Txawm tias nyob rau hauv cov neeg mob uas muaj kev pheej hmoo siab, qhov kev pheej hmoo ntawm kev mob qog nqaij hlav hauv cov qog nqaij hlav <6 hli yog <1%. Qhov kev pheej hmoo ntawm kev mob qog nqaij hlav rau cov qog nqaij hlav ntsuas 6 txog 20 hli yog txij li 8% txog 64%30. Yog li ntawd, Fleischner Society pom zoo kom muaj qhov txiav tawm ntawm 6 hli rau kev saib xyuas CT niaj hnub.29 Txawm li cas los xij, kev ntsuam xyuas kev pheej hmoo thiab kev tswj hwm ntawm cov qog nqaij hlav ntsws tsis paub meej (IPN) loj dua 6 hli tsis tau ua tiav txaus31. Kev tswj hwm tam sim no ntawm kab mob plawv yug los feem ntau yog raws li kev tos ntsoov nrog kev saib xyuas CT ntau zaus.
Raws li qhov kev kuaj mob metabolicom uas tau lees paub, peb tau ua pov thawj thawj zaug qhov kev sib tshooj ntawm cov cim qhia txog kev mob metabolicomic ntawm cov tib neeg noj qab haus huv thiab cov qog nqaij hlav benign <6 hli. Qhov kev zoo sib xws ntawm lub cev yog sib xws nrog cov kev tshawb pom CT yav dhau los uas qhov kev pheej hmoo ntawm kev mob qog nqaij hlav rau cov qog nqaij hlav <6 hli yog qis li rau cov neeg tsis muaj cov qog nqaij hlav.30 Nws yuav tsum tau sau tseg tias peb cov txiaj ntsig kuj qhia tau tias cov qog nqaij hlav benign <6 hli thiab ≥6 hli muaj kev zoo sib xws siab hauv cov qauv metabolicomic, qhia tias qhov kev txhais lus ua haujlwm ntawm benign etiology yog sib xws tsis hais qhov loj me ntawm cov qog nqaij hlav. Yog li, cov vaj huam sib luag kuaj mob niaj hnub no tuaj yeem muab ib qho kev sim ua qhov kev xeem tsis lees paub thaum cov qog nqaij hlav raug kuaj pom thawj zaug ntawm CT thiab tej zaum yuav txo qis kev saib xyuas. Tib lub sijhawm, tib lub vaj huam sib luag ntawm cov cim qhia txog kev mob metabolic tau sib txawv cov qog nqaij hlav malignant ≥6 hli hauv qhov loj me los ntawm cov qog nqaij hlav benign thiab muab cov lus kwv yees tseeb rau IPNs ntawm qhov loj me zoo sib xws thiab cov yam ntxwv tsis meej ntawm cov duab CT. Cov neeg faib tawm ntawm cov qog nqaij hlav no tau ua tiav zoo hauv kev kwv yees qhov mob qog nqaij hlav ntawm cov qog nqaij hlav ≥6 hli nrog AUC ntawm 0.927. Thaum muab ua ke, peb cov txiaj ntsig qhia tau hais tias cov cim qhia txog kev hloov pauv hauv cov ntshav hauv cov ntshav yuav qhia tau meej txog kev hloov pauv ntawm cov metabolism thaum ntxov thiab muaj txiaj ntsig zoo li cov lus qhia txog kev pheej hmoo, tsis hais qhov loj ntawm lub qog.
Qhov tseem ceeb, mob qog nqaij hlav ntsws adenocarcinoma (LUAD) thiab mob qog nqaij hlav ntsws squamous cell carcinoma (LUSC) yog cov hom mob qog nqaij hlav ntsws tsis yog me me (NSCLC). Muab hais tias LUSC muaj feem cuam tshuam nrog kev siv luam yeeb47 thiab LUAD yog qhov histology feem ntau ntawm cov qog nqaij hlav ntsws uas pom ntawm CT screening48, peb tus qauv classifier tau tsim tshwj xeeb rau theem I adenocarcinoma qauv. Wang thiab cov npoj yaig kuj tau tsom mus rau LUAD thiab tau txheeb xyuas cuaj lub cim lipid siv lipidomics los sib txawv mob qog nqaij hlav ntsws theem thaum ntxov los ntawm cov neeg noj qab haus huv17. Peb tau sim tus qauv classifier tam sim no ntawm 16 rooj plaub ntawm theem I LUSC thiab 74 lub qog nqaij hlav benign thiab pom qhov tseeb qis LUSC kwv yees (AUC 0.776), qhia tias LUAD thiab LUSC yuav muaj lawv tus kheej cov cim metabolilomic. Tseeb tiag, LUAD thiab LUSC tau pom tias txawv hauv etiology, keeb kwm biological thiab kev hloov pauv caj ces49. Yog li ntawd, lwm hom histology yuav tsum tau suav nrog rau hauv cov qauv kev cob qhia rau kev kuaj mob qog noj ntshav hauv cov pej xeem hauv cov kev pab cuam kuaj mob.
Ntawm no, peb tau txheeb xyuas rau txoj kev hloov pauv ntau tshaj plaws hauv lub ntsws adenocarcinoma piv rau cov neeg noj qab haus huv thiab cov qog nqaij hlav zoo. Xanthine thiab hypoxanthine yog cov metabolites ntawm txoj kev purine metabolic. Sib xws nrog peb cov txiaj ntsig, cov khoom nruab nrab cuam tshuam nrog purine metabolism tau nce ntxiv hauv cov ntshav lossis cov ntaub so ntswg ntawm cov neeg mob uas muaj lub ntsws adenocarcinoma piv rau cov neeg noj qab haus huv lossis cov neeg mob ntawm theem preinvasive15,50. Cov serum xanthine thiab hypoxanthine theem siab tuaj yeem cuam tshuam txog anabolism xav tau los ntawm cov hlwb qog nqaij hlav sai sai. Dysregulation ntawm glucose metabolism yog ib qho cim paub zoo ntawm kev mob qog nqaij hlav metabolism51. Ntawm no, peb tau pom qhov nce ntxiv ntawm pyruvate thiab lactate hauv pawg LA piv rau pawg HC thiab BN, uas yog sib xws nrog cov ntawv ceeb toom yav dhau los ntawm glycolytic txoj kev tsis zoo hauv cov qauv metabolome serum ntawm cov neeg mob tsis yog me me ntawm lub ntsws cancer (NSCLC) thiab cov neeg noj qab haus huv. cov txiaj ntsig yog sib xws52,53.
Qhov tseem ceeb, peb tau pom qhov sib raug zoo ntawm pyruvate thiab tryptophan metabolism hauv cov ntshav ntawm lub ntsws adenocarcinomas. Cov theem tryptophan hauv cov ntshav tau txo qis hauv pawg LA piv rau pawg HC lossis BN. Qhov nthuav yog, kev tshawb fawb loj yav dhau los siv cov neeg sawv cev yav tom ntej pom tias cov theem qis ntawm cov tryptophan ncig tau cuam tshuam nrog kev pheej hmoo ntawm mob qog noj ntshav hauv lub ntsws 54. Tryptophan yog ib qho amino acid tseem ceeb uas peb tau txais los ntawm cov zaub mov. Peb xaus lus tias cov ntshav tryptophan depletion hauv lub ntsws adenocarcinoma yuav qhia txog kev poob qis sai ntawm cov metabolite no. Nws paub zoo tias cov khoom kawg ntawm tryptophan catabolism ntawm txoj kev kynurenine yog qhov chaw ntawm de novo NAD + synthesis. Vim tias NAD + yog tsim tawm feem ntau los ntawm txoj kev cawmdim, qhov tseem ceeb ntawm NAD + hauv tryptophan metabolism hauv kev noj qab haus huv thiab kab mob tseem yuav tsum tau txiav txim siab46. Peb qhov kev tshuaj xyuas ntawm TCGA database qhia tau hais tias qhov kev qhia tawm ntawm tryptophan transporter solute transporter 7A5 (SLC7A5) tau nce ntxiv ntau hauv lub ntsws adenocarcinoma piv rau cov kev tswj hwm ib txwm thiab tau sib raug zoo nrog qhov kev qhia tawm ntawm glycolytic enzyme GAPDH. Cov kev tshawb fawb yav dhau los tau tsom mus rau lub luag haujlwm ntawm tryptophan catabolism hauv kev tiv thaiv kev tiv thaiv kab mob antitumor40,41,42. Ntawm no peb qhia tau hais tias kev txwv tsis pub tryptophan uptake los ntawm kev knockdown ntawm SLC7A5 hauv cov hlwb mob ntsws ua rau muaj kev txo qis hauv cov theem NAD cellular thiab kev ua haujlwm glycolytic. Hauv kev xaus, peb txoj kev tshawb fawb muab lub hauv paus rau kev hloov pauv hauv cov metabolism hauv cov ntshav uas cuam tshuam nrog kev hloov pauv malignant ntawm lub ntsws adenocarcinoma.
Kev hloov pauv EGFR yog cov kev hloov pauv tsav tsheb feem ntau hauv cov neeg mob uas muaj NSCLC. Hauv peb txoj kev tshawb fawb, peb pom tias cov neeg mob uas muaj kev hloov pauv EGFR (n = 41) muaj cov qauv metabolilomic zoo ib yam li cov neeg mob uas muaj hom EGFR qus (n = 31), txawm hais tias peb pom cov qib serum qis dua ntawm qee tus neeg mob EGFR mutant hauv cov neeg mob acylcarnitine. Lub luag haujlwm tsim ntawm acylcarnitines yog thauj cov pab pawg acyl los ntawm cytoplasm mus rau hauv mitochondrial matrix, ua rau oxidation ntawm cov rog acids los tsim lub zog 55. Sib xws nrog peb cov kev tshawb pom, kev tshawb fawb tsis ntev los no kuj tau txheeb xyuas cov qauv metabolilome zoo sib xws ntawm EGFR mutant thiab EGFR qus-hom qog los ntawm kev tshuaj xyuas cov metabolome thoob ntiaj teb ntawm 102 lub ntsws adenocarcinoma cov ntaub so ntswg qauv 50. Nthuav, cov ntsiab lus acylcarnitine kuj pom muaj nyob rau hauv EGFR mutant pawg. Yog li ntawd, seb qhov kev hloov pauv hauv acylcarnitine qib cuam tshuam txog EGFR-induced metabolic kev hloov pauv thiab cov hauv paus molecular txoj hauv kev yuav tsum tau kawm ntxiv.
Xaus lus, peb txoj kev tshawb fawb tsim kom muaj ib qho kev faib tawm ntawm cov metabolism hauv cov ntshav rau kev kuaj mob sib txawv ntawm cov qog hauv lub ntsws thiab tawm tswv yim rau txoj haujlwm uas tuaj yeem ua kom zoo dua qhov kev ntsuam xyuas kev pheej hmoo thiab pab txhawb kev tswj hwm kev kho mob raws li kev tshuaj ntsuam CT scan.
Txoj kev tshawb fawb no tau txais kev pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Sun Yat-sen University Cancer Hospital, Thawj Lub Tsev Kho Mob Koom Tes ntawm Sun Yat-sen University, thiab Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Zhengzhou University Cancer Hospital. Hauv cov pab pawg tshawb pom thiab kev lees paub sab hauv, 174 sera los ntawm cov tib neeg noj qab haus huv thiab 244 sera los ntawm cov nodules benign tau sau los ntawm cov tib neeg uas tau kuaj mob txhua xyoo ntawm Department of Cancer Control and Prevention, Sun Yat-sen University Cancer Center, thiab 166 benign nodules. serum. Theem I ntsws adenocarcinomas tau sau los ntawm Sun Yat-sen University Cancer Center. Hauv pawg neeg lees paub sab nraud, muaj 48 rooj plaub ntawm benign nodules, 39 rooj plaub ntawm theem I ntsws adenocarcinoma los ntawm Thawj Lub Tsev Kho Mob Koom Tes ntawm Sun Yat-sen University, thiab 24 rooj plaub ntawm theem I ntsws adenocarcinoma los ntawm Zhengzhou Cancer Hospital. Chaw Kho Mob Cancer University Sun Yat-sen kuj tau sau 16 tus neeg mob qog nqaij hlav theem I squamous cell ntsws los sim seb puas muaj peev xwm kuaj mob tau los ntawm tus neeg faib cov tshuaj metabolic (cov yam ntxwv ntawm tus neeg mob tau qhia hauv Daim Ntawv Ntxiv 5). Cov qauv los ntawm pawg neeg tshawb pom thiab pawg neeg lees paub sab hauv tau sau ntawm Lub Ib Hlis 2018 thiab Tsib Hlis 2020. Cov qauv rau pawg neeg lees paub sab nraud tau sau ntawm Lub Yim Hli 2021 thiab Kaum Hli 2022. Txhawm rau kom txo qis kev ntxub ntxaug poj niam txiv neej, kwv yees li tus lej sib npaug ntawm cov txiv neej thiab poj niam tau muab rau txhua pawg neeg. Pab Pawg Tshawb Pom thiab Pab Pawg Tshuaj Xyuas Sab Hauv. Cov poj niam txiv neej koom nrog tau txiav txim siab raws li kev tshaj tawm tus kheej. Kev pom zoo tau txais los ntawm txhua tus neeg koom nrog thiab tsis muaj kev them nyiaj tau muab. Cov neeg mob uas muaj cov qog nqaij hlav zoo yog cov uas muaj qhov qhab nia CT scan ruaj khov ntawm 2 txog 5 xyoos thaum lub sijhawm tshuaj xyuas, tshwj tsis yog 1 tus neeg mob los ntawm cov qauv lees paub sab nraud, uas tau sau ua ntej kev phais thiab kuaj mob los ntawm histopathology. Tsuas yog mob ntsws ntev xwb. Cov neeg mob ntsws adenocarcinoma tau sau ua ntej kev phais ntsws thiab lees paub los ntawm kev kuaj mob pathological. Cov qauv ntshav thaum yoo mov tau sau rau hauv cov raj sib cais serum yam tsis muaj tshuaj anticoagulants. Cov qauv ntshav tau raug clotted rau 1 teev ntawm chav tsev kub thiab tom qab ntawd centrifuged ntawm 2851 × g rau 10 feeb ntawm 4 ° C kom sau cov serum supernatant. Cov serum aliquots tau khov ntawm -80 ° C kom txog thaum rho tawm metabolite. Lub Tsev Haujlwm Saib Xyuas Kev Tiv Thaiv Kab Mob Cancer thiab Kev Tshuaj Xyuas Kev Kho Mob ntawm Sun Yat-sen University Cancer Center tau sau ib lub pas dej ntawm serum los ntawm 100 tus neeg pub dawb noj qab haus huv, suav nrog cov txiv neej thiab poj niam sib npaug hnub nyoog 40 txog 55 xyoo. Cov ntim sib npaug ntawm txhua tus qauv pub dawb tau sib xyaw, lub pas dej tau raug aliquoted thiab khaws cia ntawm -80 ° C. Cov serum sib xyaw tau siv ua cov ntaub ntawv siv rau kev tswj xyuas zoo thiab kev ua qauv ntawm cov ntaub ntawv.
Cov qauv serum thiab cov qauv kuaj tau yaj thiab cov metabolites tau rho tawm siv txoj kev rho tawm ua ke (MTBE/methanol/dej) 56. Hauv luv luv, 50 μl ntawm serum tau sib xyaw nrog 225 μl ntawm cov dej khov-txias methanol thiab 750 μl ntawm cov dej khov-txias methyl tert-butyl ether (MTBE). Do cov khoom sib xyaw thiab incubate rau ntawm dej khov rau 1 teev. Cov qauv tau sib xyaw thiab vortex sib xyaw nrog 188 μl ntawm MS-qib dej uas muaj cov qauv sab hauv (13C-lactate, 13C3-pyruvate, 13C-methionine, thiab 13C6-isoleucine, yuav los ntawm Cambridge Isotope Laboratories). Cov khoom sib xyaw ces tau centrifuged ntawm 15,000 × g rau 10 feeb ntawm 4 ° C, thiab theem qis dua tau hloov mus rau hauv ob lub raj (125 μL txhua) rau LC-MS kev tshuaj xyuas hauv hom zoo thiab tsis zoo. Thaum kawg, cov qauv tau evaporated kom qhuav hauv lub tshuab nqus tsev ceev ceev.
Cov metabolites qhuav tau rov ua dua tshiab hauv 120 μl ntawm 80% acetonitrile, vortexed rau 5 feeb, thiab centrifuged ntawm 15,000 × g rau 10 feeb ntawm 4 ° C. Supernatants tau hloov mus rau hauv lub raj mis iav amber nrog microinserts rau kev tshawb fawb metabolomics. Untargeted metabolomics kev tshuaj xyuas ntawm lub ultra-performance kua chromatography-high-resolution mass spectrometry (UPLC-HRMS) platform. Metabolites tau sib cais siv Dionex Ultimate 3000 UPLC system thiab ACQUITY BEH Amide kem (2.1 × 100 mm, 1.7 μm, Waters). Hauv hom ion zoo, cov theem txawb yog 95% (A) thiab 50% acetonitrile (B), txhua tus muaj 10 mmol / L ammonium acetate thiab 0.1% formic acid. Hauv hom tsis zoo, cov theem txawb A thiab B muaj 95% thiab 50% acetonitrile, feem, ob theem muaj 10 mmol/L ammonium acetate, pH = 9. Qhov kev pab cuam gradient yog raws li nram no: 0–0.5 feeb, 2% B; 0.5–12 feeb, 2–50% B; 12–14 feeb, 50–98% B; 14–16 feeb, 98% B; 16–16.1. feeb, 98 –2% B; 16.1–20 feeb, 2% B. Lub kem tau khaws cia ntawm 40°C thiab cov qauv ntawm 10°C hauv lub autosampler. Tus nqi ntws yog 0.3 ml/min, qhov ntim txhaj tshuaj yog 3 μl. Ib lub Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific) nrog lub electrospray ionization (ESI) qhov chaw tau ua haujlwm hauv hom scan tag nrho thiab txuas nrog hom kev saib xyuas ddMS2 kom sau cov ntaub ntawv ntau. Cov kev teeb tsa MS tau teeb tsa raws li nram no: lub zog tsuag +3.8 kV / - 3.2 kV, capillary kub 320 ° C, roj shielding 40 arb, roj auxiliary 10 arb, probe heater kub 350 ° C, scanning ntau 70–1050 m / h, kev daws teeb meem. 70 000. Cov ntaub ntawv tau txais los ntawm kev siv Xcalibur 4.1 (Thermo Fisher Scientific).
Txhawm rau ntsuam xyuas qhov zoo ntawm cov ntaub ntawv, cov qauv tswj kev zoo (QC) tau tsim los ntawm kev tshem tawm 10 μL aliquots ntawm supernatant los ntawm txhua tus qauv. Rau qhov kev txhaj tshuaj tswj kev zoo tau raug soj ntsuam thaum pib ntawm cov txheej txheem analytical los ntsuam xyuas qhov ruaj khov ntawm UPLC-MS system. Cov qauv tswj kev zoo tom qab ntawd raug qhia tsis tu ncua rau hauv pawg. Tag nrho 11 pawg ntawm cov qauv serum hauv txoj kev tshawb fawb no tau raug soj ntsuam los ntawm LC-MS. Cov aliquots ntawm cov dej sib xyaw serum los ntawm 100 tus neeg pub dawb noj qab haus huv tau siv ua cov ntaub ntawv siv hauv cov pawg sib txawv los saib xyuas cov txheej txheem rho tawm thiab kho rau cov teebmeem batch-to-batch. Kev tshuaj xyuas metabolomics tsis tsom mus rau ntawm pawg neeg tshawb pom, pawg neeg lees paub sab hauv, thiab pawg neeg lees paub sab nraud tau ua tiav ntawm Metabolomics Center ntawm Sun Yat-sen University. Lub chaw soj nstuam sab nraud ntawm Guangdong University of Technology Analysis and Testing Center kuj tau tshuaj xyuas 40 cov qauv los ntawm pawg neeg sab nraud los sim qhov kev ua tau zoo ntawm tus qauv classifier.
Tom qab rho tawm thiab rov ua dua, kev ntsuas qhov tseeb ntawm cov metabolites hauv cov ntshav tau ntsuas siv cov kua chromatography-tandem mass spectrometry (Agilent 6495 triple quadrupole) nrog lub electrospray ionization (ESI) qhov chaw hauv ntau hom kev tshuaj xyuas kev ua haujlwm (MRM). Ib kem ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Dej) tau siv los cais cov metabolites. Lub theem txawb muaj 90% (A) thiab 5% acetonitrile (B) nrog 10 mmol / L ammonium acetate thiab 0.1% ammonia kua. Qhov kev pab cuam gradient yog raws li nram no: 0–1.5 feeb, 0% B; 1.5–6.5 feeb, 0–15% B; 6.5–8 feeb, 15% B; 8–8.5 feeb, 15%–0% B; 8.5–11.5 feeb, 0%B. Lub kem tau khaws cia ntawm 40 ° C thiab cov qauv ntawm 10 ° C hauv lub autosampler. Tus nqi ntws yog 0.3 mL / feeb thiab qhov ntim txhaj tshuaj yog 1 μL. Cov kev teeb tsa MS tau teeb tsa raws li nram no: capillary voltage ± 3.5 kV, nebulizer siab 35 psi, sheath gas flow 12 L / min, sheath gas kub 350 ° C, drying gas kub 250 ° C, thiab drying gas flow 14 l / min. Cov kev hloov pauv MRM ntawm tryptophan, pyruvate, lactate, hypoxanthine thiab xanthine yog 205.0–187.9, 87.0–43.4, 89.0–43.3, 135.0–92.3 thiab 151.0–107.9 feem. Cov ntaub ntawv tau sau los ntawm kev siv Mass Hunter B.07.00 (Agilent Technologies). Rau cov qauv serum, tryptophan, pyruvate, lactate, hypoxanthine, thiab xanthine tau ntsuas siv cov kab calibration ntawm cov txheej txheem sib tov. Rau cov qauv cell, cov ntsiab lus tryptophan tau normalized rau sab hauv tus qauv thiab cov protein cell.
Kev rho tawm qhov siab tshaj plaws (m/z thiab lub sijhawm khaws cia (RT)) tau ua tiav siv Compound Discovery 3.1 thiab TraceFinder 4.0 (Thermo Fisher Scientific). Txhawm rau tshem tawm qhov sib txawv ntawm cov pawg, txhua qhov siab tshaj plaws ntawm cov qauv kuaj tau faib los ntawm qhov siab tshaj plaws ntawm cov khoom siv los ntawm tib pawg kom tau txais qhov muaj ntau ntxiv. Qhov sib txawv ntawm cov qauv sab hauv ua ntej thiab tom qab kev ntsuas tus qauv tau qhia hauv Supplementary Table 6. Qhov sib txawv ntawm ob pawg tau piav qhia los ntawm qhov tshwm sim tsis tseeb (FDR <0.05, Wilcoxon kos npe qib xeem) thiab kev hloov pauv (>1.2 lossis <0.83). Cov ntaub ntawv MS nyoos ntawm cov yam ntxwv rho tawm thiab cov ntaub ntawv MS kho serum tau qhia hauv Supplementary Data 1 thiab Supplementary Data 2, raws li. Kev piav qhia siab tshaj plaws tau ua tiav raws li plaub qib kev txheeb xyuas, suav nrog cov metabolites txheeb xyuas, cov tshuaj sib xyaw ua ke, cov chav kawm sib xyaw ua ke, thiab cov tshuaj tsis paub 22. Raws li kev tshawb nrhiav hauv database hauv Compound Discovery 3.1 (mzCloud, HMDB, Chemspider), cov tshuaj lom neeg nrog MS/MS phim cov qauv lees paub lossis cov lus piav qhia sib phim hauv mzCloud (qhab nia > 85) lossis Chemspider thaum kawg tau xaiv ua cov khoom nruab nrab ntawm cov metabolome sib txawv. Cov lus piav qhia siab tshaj plaws rau txhua qhov feature tau suav nrog hauv Cov Ntaub Ntawv Ntxiv 3. MetaboAnalyst 5.0 tau siv rau kev tshuaj xyuas univariate ntawm cov metabolite ntau ntau. MetaboAnalyst 5.0 kuj tau soj ntsuam KEGG txoj kev tshuaj xyuas kev nplua nuj raws li cov metabolites sib txawv. Kev tshuaj xyuas cov khoom tseem ceeb (PCA) thiab kev tshuaj xyuas qhov sib txawv tsawg kawg nkaus (PLS-DA) tau tshuaj xyuas siv cov pob software ropls (v.1.26.4) nrog stack normalization thiab autoscaling. Tus qauv metabolite biomarker zoo tshaj plaws rau kev kwv yees nodule malignancy tau tsim los ntawm kev siv binary logistic regression nrog qhov tsawg kawg nkaus shrinkage thiab xaiv tus neeg teb xov tooj (LASSO, R pob v.4.1-3). Kev ua tau zoo ntawm tus qauv kev sib txawv hauv cov txheej txheem kuaj pom thiab kev lees paub tau piav qhia los ntawm kev kwv yees AUC raws li ROC kev tshuaj xyuas raws li pob pROC (v.1.18.0.). Qhov kev txiav tawm zoo tshaj plaws tau txais raws li qhov siab tshaj plaws Youden index ntawm tus qauv (kev nkag siab + qhov tshwj xeeb - 1). Cov qauv nrog cov nqi tsawg dua lossis ntau dua qhov txwv yuav raug kwv yees tias yog cov qog nqaij hlav thiab lub ntsws adenocarcinoma, raws li.
Cov qe ntshav A549 (#CCL-185, American Type Culture Collection) tau cog rau hauv F-12K medium uas muaj 10% FBS. Cov kab lus luv luv hairpin RNA (shRNA) uas tsom mus rau SLC7A5 thiab cov kev tswj tsis yog targeting (NC) tau ntxig rau hauv lentiviral vector pLKO.1-puro. Cov kab lus antisense ntawm shSLC7A5 yog raws li nram no: Sh1 (5′-GGAGAAACCTGATGAACAGTT-3′), Sh2 (5′-GCCGTGGACTTCGGGAACTAT-3′). Cov tshuaj tua kab mob rau SLC7A5 (#5347) thiab tubulin (#2148) tau yuav los ntawm Cell Signaling Technology. Cov tshuaj tua kab mob rau SLC7A5 thiab tubulin tau siv ntawm qhov dilution ntawm 1:1000 rau Western blot analysis.
Qhov Seahorse XF Glycolytic Stress Test ntsuas cov theem ntawm extracellular acidification (ECAR). Hauv qhov kev ntsuam xyuas, glucose, oligomycin A, thiab 2-DG tau muab tshuaj sib law liag los kuaj lub peev xwm glycolytic ntawm cov hlwb raws li ntsuas los ntawm ECAR.
Cov qe ntshav A549 uas tau hloov pauv nrog kev tswj tsis yog targeting (NC) thiab shSLC7A5 (Sh1, Sh2) tau plated ib hmos hauv cov tais diav 10 cm uas muaj txoj kab uas hla. Cov metabolites ntawm cov qe ntshav tau rho tawm nrog 1 ml ntawm dej khov txias 80% aqueous methanol. Cov qe ntshav hauv cov kua methanol tau khawb tawm, sau rau hauv lub raj tshiab, thiab centrifuged ntawm 15,000 × g rau 15 feeb ntawm 4°C. Sau 800 µl ntawm supernatant thiab ziab siv lub tshuab nqus tsev ceev ceev. Cov metabolite pellets qhuav tau raug soj ntsuam rau cov theem tryptophan siv LC-MS/MS raws li tau piav qhia saum toj no. Cov theem Cellular NAD(H) hauv A549 qe ntshav (NC thiab shSLC7A5) tau ntsuas siv cov khoom siv NAD+/NADH colorimetric (#K337, BioVision) raws li cov lus qhia ntawm cov chaw tsim khoom. Cov theem Protein tau ntsuas rau txhua tus qauv kom ua kom cov metabolites zoo li qub.
Tsis muaj cov txheej txheem suav lej siv los txiav txim siab qhov loj ntawm cov qauv. Cov kev tshawb fawb metabolomics yav dhau los uas tsom mus rau kev tshawb pom biomarker15,18 tau raug suav hais tias yog cov qauv rau kev txiav txim siab qhov loj me, thiab piv rau cov ntawv ceeb toom no, peb cov qauv txaus. Tsis muaj cov qauv raug tshem tawm ntawm pawg neeg kawm. Cov qauv ntshav tau raug xaiv rau pawg tshawb pom (306 rooj plaub, 74.6%) thiab pawg lees paub sab hauv (104 rooj plaub, 25.4%) rau kev tshawb fawb metabolomics tsis tsom. Peb kuj tau xaiv 70 rooj plaub los ntawm txhua pawg los ntawm cov txheej txheem tshawb pom rau kev tshawb fawb metabolomics tsom. Cov kws tshawb nrhiav tau dig muag rau kev faib pawg thaum lub sijhawm sau thiab tshuaj xyuas cov ntaub ntawv LC-MS. Kev tshuaj xyuas cov ntaub ntawv metabolomics thiab kev sim cell tau piav qhia hauv cov txiaj ntsig, Cov Lus Cim, thiab Cov Txheej Txheem. Kev ntsuas ntawm cellular tryptophan, NADT, thiab glycolytic kev ua haujlwm tau ua peb zaug ywj pheej nrog cov txiaj ntsig zoo sib xws.
Yog xav paub ntxiv txog kev tsim txoj kev tshawb fawb, mus saib Natural Portfolio Report Abstract uas cuam tshuam nrog tsab xov xwm no.
Cov ntaub ntawv MS raw ntawm cov yam ntxwv rho tawm thiab cov ntaub ntawv MS normalized ntawm cov serum siv tau qhia nyob rau hauv Cov Ntaub Ntawv Ntxiv 1 thiab Cov Ntaub Ntawv Ntxiv 2, raws li. Cov lus piav qhia txog qhov siab tshaj plaws rau cov yam ntxwv sib txawv tau nthuav tawm nyob rau hauv Cov Ntaub Ntawv Ntxiv 3. Cov ntaub ntawv LUAD TCGA tuaj yeem rub tawm los ntawm https://portal.gdc.cancer.gov/. Cov ntaub ntawv nkag rau kev kos duab daim duab tau muab rau hauv cov ntaub ntawv qhov chaw. Cov ntaub ntawv qhov chaw tau muab rau tsab xov xwm no.
Pawg Kawm Txog Kev Tshuaj Xyuas Lub Ntsws Hauv Tebchaws, thiab lwm yam. Txo kev tuag ntawm mob qog noj ntshav hauv lub ntsws nrog kev siv tshuaj xoo hluav taws xob tsawg. Northern England. J. Med. 365, 395–409 (2011).
Kramer, BS, Berg, KD, Aberle, DR thiab Prophet, PC Kev tshuaj ntsuam mob qog noj ntshav hauv lub ntsws siv cov koob tshuaj helical CT qis: cov txiaj ntsig los ntawm National Lung Screening Study (NLST). J. Med. Screen 18, 109–111 (2011).
De Koning, HJ, thiab lwm tus. Txo kev tuag los ntawm mob qog noj ntshav hauv lub ntsws nrog kev tshuaj ntsuam CT volumetric hauv kev sim tshuaj randomized. Northern England. J. Med. 382, 503–513 (2020).
Lub sijhawm tshaj tawm: Cuaj hlis-18-2023