ʻO ka ʻike ʻokoʻa o nā nodules pulmonary i ʻike ʻia e ka computed tomography (CT) he paʻakikī i ka hana lapaʻau.Ma ʻaneʻi, ʻike mākou i ka metabolome honua o 480 serum samples, me ka hoʻomalu olakino, benign lung nodules, a me ka pae I lung adenocarcinoma.Hōʻike nā Adenocarcinomas i nā profile metabolomic kūʻokoʻa, ʻoiai ʻo nā nodules benign a me nā kānaka olakino he like kiʻekiʻe i nā profile metabolomic.I loko o ka hui ʻike (n = 306), ua ʻike ʻia kahi pūʻulu o 27 metabolites e hoʻokaʻawale i waena o nā nodules maikaʻi a maikaʻi ʻole.ʻO ka AUC o ka hoʻohālike hoʻokae i loko o ka hōʻoia kūloko (n = 104) a me ka hōʻoia waho (n = 111) pūʻulu he 0.915 a me 0.945.Ua hōʻike ʻia ka loiloi ala i ka hoʻonui ʻana i nā metabolites glycolytic e pili ana me ka hoʻemi ʻana i ka tryptophan i loko o ka serum adenocarcinoma māmā i hoʻohālikelike ʻia me nā nodules maikaʻi a me nā mana olakino, a manaʻo ʻia ʻo tryptophan uptake e hoʻoikaika i ka glycolysis i loko o nā pūnaʻi maʻi kanesa.Hōʻike kā mākou noiʻi i ka waiwai o nā biomarkers metabolite serum i ka loiloi ʻana i ka pilikia o nā nodules pulmonary i ʻike ʻia e CT.
He mea koʻikoʻi ka hoʻomaʻamaʻa mua ʻana e hoʻomaikaʻi i nā helu ola no nā maʻi maʻi kanesa.Ua hōʻike nā hualoaʻa mai ka US National Lung Cancer Screening Trial (NLST) a me ka European NELSON Study i ka nānā ʻana me ka haʻahaʻa haʻahaʻa computed tomography (LDCT) hiki ke hōʻemi nui i ka make maʻi maʻi maʻi maʻi i nā hui kiʻekiʻe 1,2,3.Mai ka hoʻohana nui ʻana o ka LDCT no ka nānā ʻana i ka maʻi kanesa, ua hoʻomau ka piʻi ʻana o nā ʻike radiographic kūikawā o nā nodules asymptomatic pulmonary nodules.Ua wehewehe ʻia nā nodule pulmonary ma ke ʻano he opacities focal a hiki i 3 knm ke anawaena 5.Ke pilikia nei mākou i ka loiloi ʻana i ka nui o nā nodules pulmonary i ʻike ʻia ma ka LDCT.ʻO nā palena o CT hiki ke alakaʻi i nā hoʻokolohua hahai pinepine a me nā hopena maikaʻi ʻole, e alakaʻi ana i ka hana pono ʻole a me ka mālama ʻana6.No laila, pono e hoʻomohala i nā biomarkers hilinaʻi a pono e hoʻomaopopo pono i ka maʻi kanesa o ka maʻi maʻi i ka wā mua a hoʻokaʻawale i nā nodules maikaʻi loa i ka ʻike mua ʻana 7.
ʻO ka hoʻopaʻa ʻana i ka molecular analysis o ke koko (serum, plasma, peripheral blood mononuclear cell), me nā genomics, proteomics a i ʻole DNA methylation8,9,10, ua alakaʻi i ka ulu ʻana o ka hoihoi i ka loaʻa ʻana o nā biomarkers diagnostic no ka maʻi kanesa.Eia nō naʻe, ke ana ʻana o ka metabolomics i nā huahana hope cellular i hoʻopili ʻia e nā hana endogenous a exogenous a no laila ua noi ʻia e wānana i ka hoʻomaka ʻana o ka maʻi a me ka hopena.ʻO Liquid chromatography-tandem mass spectrometry (LC-MS) kahi ala i hoʻohana nui ʻia no nā haʻawina metabolomics ma muli o kona kiʻekiʻe kiʻekiʻe a me ka nui dynamic range, hiki ke uhi i nā metabolites me nā waiwai physicochemical like ʻole11,12,13.ʻOiai ua hoʻohana ʻia ka loiloi metabolomic honua o ka plasma / serum e ʻike i nā biomarkers e pili ana i ka maʻi maʻi maʻi maʻi maʻi diagnostic14,15,16,17 a me ka maikaʻi o ka mālama ʻana, 18 serum metabolite classifiers e hoʻokaʻawale i waena o nā nodules māmā a me ka malignant e aʻo nui ʻia.-ka noiʻi nui.
ʻO Adenocarcinoma a me squamous cell carcinoma nā ʻano subtypes ʻelua o ka maʻi maʻi ʻaʻai liʻiliʻi liʻiliʻi (NSCLC).Hōʻike nā hoʻāʻo hōʻike CT like ʻole ʻo adenocarcinoma ka ʻano histological maʻamau o ka maʻi maʻi maʻi maʻi 1,19,20,21.Ma kēia haʻawina, ua hoʻohana mākou i ka ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) e hana i ka metabolomics analysis ma ka huina o 695 serum samples, me nā mana olakino, benign pulmonary nodules, a me CT-detected ≤3 cm.ʻO ka nānā ʻana no ka Stage I lung adenocarcinoma.Ua ʻike mākou i kahi papa o nā metabolites serum e hoʻokaʻawale i ka adenocarcinoma māmā mai nā nodules maikaʻi a me nā mana olakino.Ua hōʻike ʻia ka nānā ʻana i ka hoʻonui ʻana i ke ala ʻo ka tryptophan a me ka metabolism glucose he mau loli maʻamau i ka adenocarcinoma māmā i hoʻohālikelike ʻia me nā nodules maikaʻi a me nā mana olakino.ʻO ka mea hope loa, ua hoʻokumu mākou a hōʻoia i kahi mea hoʻohālikelike metabolic serum me ka kikoʻī kiʻekiʻe a me ka naʻau e hoʻokaʻawale i waena o nā nodules malignant a me benign pulmonary nodules i ʻike ʻia e LDCT, hiki ke kōkua i ka maʻi ʻokoʻa mua a me ka loiloi pilikia.
Ma ke aʻo ʻana i kēia manawa, ua hōʻiliʻili ʻia nā laʻana serum pili i ke kāne a me ka makahiki mai 174 mau mana olakino, 292 mau maʻi me nā nodules pulmonary benign, a me 229 mau maʻi me ka pae I lung adenocarcinoma.Hōʻike ʻia nā hiʻohiʻona demographic o nā kumuhana 695 ma ka Papa Pākuʻi 1.
E like me ka mea i hōʻike ʻia ma ka Figure 1a, he 480 serum samples, me 174 health control (HC), 170 benign nodules (BN), a me 136 stage I lung adenocarcinoma (LA) samples, i hōʻiliʻili ʻia ma Sun Yat-sen University Cancer Center.ʻO ka ʻimi ʻimi no ka hoʻopili ʻana i ka metabolomic ʻole me ka hoʻohana ʻana i ka ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS).E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 1, ua ʻike ʻia nā metabolites ʻokoʻa ma waena o LA a me HC, LA a me BN e hoʻokumu i kahi kumu hoʻohālike a e ʻimi hou aku i ka nānā ʻana i ke ala ʻokoʻa.ʻO 104 mau laʻana i hōʻiliʻili ʻia e Sun Yat-sen University Cancer Center a me 111 mau hōʻailona i hōʻiliʻili ʻia e ʻelua mau haukapila ʻē aʻe i hoʻopaʻa ʻia i loko a me waho.
He heluna kanaka noiʻi ma ka hui ʻike i loaʻa i ka nānā ʻana i ka serum metabolomics honua me ka hoʻohana ʻana i ka ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS).b ʻAhaʻi hapa liʻiliʻi loa discriminant analysis (PLS-DA) o ka huina metabolome o 480 serum laʻana mai ka pūʻulu noiʻi, me nā mana olakino (HC, n = 174), nā nodules maikaʻi (BN, n = 170), a me ka pae I lung adenocarcinoma (Los Angeles, n = 136).+ ESI, ke ʻano hoʻoheheʻe electrospray maikaʻi, -ESI, ke ʻano hoʻoheheʻe electrospray maikaʻi ʻole.c–e Hōʻike ʻia nā metabolites me ka nui ʻokoʻa i loko o ʻelua pūʻulu i hāʻawi ʻia (hōʻike ʻia ʻelua-tailed Wilcoxon i hoʻopaʻa inoa ʻia, helu kuhi hewa i hoʻololi ʻia i ka p waiwai, FDR <0.05) i hōʻike ʻia i ka ʻulaʻula (fold change > 1.2) a me ka uliuli (fold change <0.83) .) hōʻike ʻia ma ke kiʻi lua pele.f Hierarchical clustering heat map e hōʻike ana i nā ʻokoʻa nui o ka helu o nā metabolites annotated ma waena o LA a me BN.Hāʻawi ʻia ka ʻikepili kumu ma ke ʻano o nā faila ʻikepili kumu.
ʻO ka nui o ka metabolome serum o 174 HC, 170 BN a me 136 LA i loko o ka hui ʻike i kālai ʻia me ka hoʻohana ʻana i ka nānā ʻana UPLC-HRMS.Hōʻike mua mākou i ka hōʻuluʻulu ʻana o ka mana maikaʻi (QC) ma ke kikowaena o kahi kumu hoʻohālike kumu ʻole i mālama ʻia (PCA), e hōʻoia ana i ka paʻa o ka hana o kēia haʻawina (Supplementary Figure 2).
E like me ka mea i hōʻike ʻia ma ka ʻāpana liʻiliʻi liʻiliʻi loa-discriminant analysis (PLS-DA) ma ke Kiʻi 1 b, ua ʻike mākou aia nā ʻokoʻa maopopo ma waena o LA a me BN, LA a me HC i nā ʻano hana electrospray ionization maikaʻi (+ESI) a me nā ʻano ʻino (-ESI). .kaawale.Eia naʻe, ʻaʻohe ʻokoʻa koʻikoʻi i loaʻa ma waena o BN a me HC ma + ESI a me -ESI kūlana.
Ua loaʻa iā mākou he 382 mau hiʻohiʻona ʻokoʻa ma waena o LA a me HC, 231 mau hiʻohiʻona ʻokoʻa ma waena o LA a me BN, a me 95 mau hiʻohiʻona ʻokoʻa ma waena o BN a me HC (Wilcoxon i hoʻopaʻa inoa ʻia, FDR <0.05 a me nā hoʻololi lehulehu> 1.2 a i ʻole <0.83) (Figure .1c-e )..Ua hōʻike hou ʻia nā kiʻekiʻe (Supplementary Data 3) e kūʻē i kahi waihona (mzCloud/HMDB/Chemspider library) e ka waiwai m / z, ka manawa paʻa a me ka huli ʻana i ka spectrum mass fragmentation (nā kikoʻī i wehewehe ʻia ma ka ʻāpana Methods) 22.ʻO ka mea hope loa, ua ʻike ʻia nā metabolites annotated 33 a me 38 me nā ʻokoʻa nui o ka nui no LA versus BN (Figure 1f and Supplementary Table 2) a me LA versus HC (Supplementary Figure 3 and Supplementary Table 2), kēlā me kēia.I ka hoʻohālikelike ʻana, ʻo 3 mau metabolites me nā ʻokoʻa koʻikoʻi i ka nui i ʻike ʻia ma BN a me HC (Supplementary Table 2), e kūlike me ka overlap ma waena o BN a me HC ma PLS-DA.Hoʻopili kēia mau metabolites ʻokoʻa i kahi ākea o nā biochemicals (Supplementary Figure 4).Hoʻohui pū ʻia, hōʻike kēia mau hopena i nā loli nui i ka serum metabolome e hōʻike ana i ka hoʻololi ʻino ʻana o ka maʻi maʻi maʻi maʻi maʻi mua i hoʻohālikelike ʻia me nā nodules māmā a i ʻole nā mea olakino.I kēia manawa, ʻo ka like o ka serum metabolome o BN a me HC e hōʻike ana e hiki i nā nodules pulmonary benign ke kaʻana like i nā ʻano olaola me nā kānaka olakino.Ma muli o ka maʻamau o ka epidermal growth factor receptor (EGFR) gene mutations i loko o ka māmā adenocarcinoma subtype 23, ua ʻimi mākou e hoʻoholo i ka hopena o ka hoʻololi ʻana o ka mea hoʻokele i ka serum metabolome.A laila nānā mākou i ka ʻikepili metabolomic holoʻokoʻa o nā hihia 72 me ke kūlana EGFR i ka pūʻulu adenocarcinoma māmā.ʻO ka mea mahalo, ua ʻike mākou i nā ʻano like like ma waena o nā maʻi maʻi mutant EGFR (n = 41) a me nā maʻi maʻi ʻano hihiu EGFR (n = 31) i ka nānā ʻana PCA (Supplementary Figure 5a).Eia nō naʻe, ua ʻike mākou i nā metabolites 7 nona ka nui i hoʻololi nui ʻia i nā maʻi me ka mutation EGFR i hoʻohālikelike ʻia me nā maʻi me ka wild-type EGFR (t test, p <0.05 a me ka loli hoʻololi> 1.2 a i ʻole <0.83) (Supplementary Figure 5b).ʻO ka hapa nui o kēia mau metabolites (5 mai ka 7) he acylcarnitines, kahi mea nui i nā ala hoʻoheheʻe momona momona.
E like me ka mea i hōʻike ʻia ma ke kaʻina hana i hōʻike ʻia ma ka Figure 2 a, ua loaʻa nā biomarkers no ka hoʻohālikelike nodule me ka hoʻohana ʻana i nā mea hoʻohana liʻiliʻi loa a me ke koho ʻana ma muli o 33 mau metabolites ʻokoʻa i ʻike ʻia ma LA (n = 136) a me BN (n = 170).ʻO ka hui maikaʻi loa o nā mea hoʻololi (LASSO) - binary logistic regression model.Ua hoʻohana ʻia ka hōʻoia cross-validation he ʻumi e hoʻāʻo ai i ka hilinaʻi o ke kumu hoʻohālike.Hoʻololi ʻia ke koho ʻokoʻa a me ka hoʻoponopono ʻana i nā ʻāpana e ka hoʻopaʻi hoʻonui ʻia me ka parameter λ24.Ua hoʻokō hou ʻia ka loiloi metabolomics honua ma ke kūʻokoʻa i ka hōʻoia kūloko (n = 104) a me ka hōʻoia waho (n = 111) mau pūʻulu e hoʻāʻo ai i ka hana hoʻohālikelike ʻana o ke ʻano hoʻohālikelike.ʻO ka hopena, ua ʻike ʻia nā metabolites 27 i ka hoʻonohonoho ʻike ʻia ʻo ke kumu hoʻohālikelike maikaʻi loa me ka waiwai AUC mean nui loa (Fig. 2b), ma waena o 9 i hoʻonui i ka hana a me 18 i hoʻemi i ka hana ma LA i hoʻohālikelike ʻia me BN (Fig. 2c).
Ka holo hana no ke kukulu ana i ka pulmonary nodule classifier, me ke koho ana i ka papa maikai loa o ka serum metabolites i ka mea i ike ia me ka hoohana ana i ke kumu hoolike hoololi logistic binary ma o ka hooiaio kea he umi a me ka loiloi ana i ka hana wanana i loko a me waho hoonohonoho hooia.b Nā helu helu hōʻoia o LASSO regression model no ke koho ʻana i ka biomarker metabolic.ʻO nā helu i hāʻawi ʻia ma luna nei ka helu awelika o nā biomarkers i koho ʻia ma kahi λ i hāʻawi ʻia.Hōʻike ka laina kiko ʻulaʻula i ka waiwai AUC maʻamau ma ka lambda pili.Hōʻike nā pahu kuhi hina hina i ka liʻiliʻi a me ka nui o nā waiwai AUC.Hōʻike ka laina kiko i ke kumu hoʻohālike maikaʻi loa me nā biomarkers koho 27.AUC, wahi ma lalo o ka pihi hana hoʻohana (ROC).c Hoʻololi hoʻololi o 27 metabolites i koho ʻia i ka hui LA i hoʻohālikelike ʻia me ka pūʻulu BN ma ka hui ʻike.kolamu ʻulaʻula - hoʻāla.He emi ke kolamu uliuli.d–f Receiver operating characteristic (ROC) curves e hōʻike ana i ka mana o ke kŘkohu hoʻokae ma muli o 27 metabolite hui pū ʻana i ka ʻike, loko, a me waho.Hāʻawi ʻia ka ʻikepili kumu ma ke ʻano o nā faila ʻikepili kumu.
Ua hana ʻia kahi ʻano wānana e pili ana i nā coefficient regression paona o kēia mau metabolites 27 (Paʻi Hoʻohui 3).ʻO ka loiloi ROC e pili ana i kēia mau metabolites 27 i hāʻawi i kahi wahi ma lalo o ka curve (AUC) waiwai o 0.933, ʻike ʻia ka ʻike hui ʻo 0.868, a ʻo 0.859 ka kikoʻī (Fig. 2d).I kēia manawa, ma waena o ka 38 annotated differential metabolites ma waena o LA a me HC, ua loaʻa kahi set o 16 metabolites i kahi AUC o 0.902 me kahi sensitivity o 0.801 a me kahi kikoʻī o 0.856 i ka hoʻokaʻawale ʻana iā LA mai HC (Supplementary Figure 6a-c).Ua hoʻohālikelike ʻia nā waiwai AUC e pili ana i nā paepae hoʻololi like ʻole no nā metabolites ʻokoʻa.Ua ʻike mākou ua hana maikaʻi ke ʻano hoʻohālikelike i ka hoʻokaʻawale ʻana ma waena o LA a me BN (HC) i ka wā i hoʻonohonoho ʻia ai ka pae hoʻololi i ka 1.2 versus 1.5 a i ʻole 2.0 (Supplementary Figure 7a,b).ʻO ke kumu hoʻohālikelike, e pili ana i nā pūʻulu metabolite 27, ua hōʻoia hou ʻia i nā cohorts i loko a me waho.ʻO ka AUC he 0.915 (ʻike 0.867, kiko'ī 0.811) no ka hōʻoia o loko a me 0.945 (ʻike 0.810, kiko'ī 0.979) no ka hōʻoia waho (Fig. 2e, f).No ka loiloi i ka pono interlaboratory, 40 mau laʻana mai ka cohort waho i kālailai ʻia i loko o kahi keʻena o waho e like me ka wehewehe ʻana ma ka ʻāpana Methods.Ua loaʻa ka pololei o ka hoʻokaʻawale ʻana i kahi AUC o 0.925 (Hei 8).No ka mea, ʻo ka lung squamous cell carcinoma (LUSC) ka lua o ka subtype maʻamau o ka non-small cell lung cancer (NSCLC) ma hope o ka lung adenocarcinoma (LUAD), ua hoʻāʻo pū mākou i ka pono pono kūpono o nā profile metabolic.BN a me 16 mau hihia o LUSC.ʻO ka AUC o ka hoʻokae ma waena o LUSC a me BN he 0.776 (Supplementary Figure 9), e hōʻike ana i ka hiki ʻole ke hoʻohālikelike i ka hoʻokae ma waena o LUAD a me BN.
Ua hōʻike nā haʻawina i ka nui o nā nodules ma nā kiʻi CT e hoʻopili maikaʻi ʻia me ka likelihood o ka maʻi maʻi a hoʻomau i ka hoʻoholo nui o ka nodule treatment25,26,27.ʻO kaʻikepili o kaʻikepili mai ka pūʻulu nui o ka NELSON screening study i hōʻike i ka pilikia o ka maʻiʻino i nā kumuhana me nā nodes <5 mm ua like like me nā kumuhana me kaʻole o nā nodes 28.No laila, ʻo ka liʻiliʻi liʻiliʻi e koi ana i ka nānā ʻana i ka CT maʻamau ʻo 5 mm, e like me ka mea i ʻōlelo ʻia e ka British Thoracic Society (BTS), a me 6 mm, e like me ka ʻōlelo a ka Fleischner Society 29.Eia nō naʻe, ʻoi aku ka nui o nā nodules ma mua o 6 mm a me ka ʻole o nā hiʻohiʻona maikaʻi ʻole, i kapa ʻia ʻo indeterminate pulmonary nodules (IPN), he mea paʻakikī nui i ka loiloi a me ka hoʻokele ʻana i ka hana lapaʻau30,31.Ua nānā hou mākou inā ua hoʻololi ka nui o ka nodule i nā inoa metabolomic me ka hoʻohana ʻana i nā hōʻailona pooled mai ka ʻike a me nā cohorts hōʻoia kūloko.Ke nānā nei i nā biomarkers i hoʻopaʻa ʻia i ka 27, ua hoʻohālikelike mua mākou i nā profiles PCA o HC a me BN sub-6 mm metabolomes.Ua ʻike mākou ʻo ka hapa nui o nā helu ʻikepili no HC a me BN i uhi ʻia, e hōʻike ana ua like nā pae metabolite serum i nā hui ʻelua (Fig. 3a).Ua mālama ʻia nā palapala hiʻohiʻona ma waena o nā pae nui like ʻole ma BN a me LA (Fig. 3b, c), ʻoiai ua ʻike ʻia kahi kaʻawale ma waena o nā nodules malignant a me benign ma ka laulā 6-20 mm (Fig. 3d).He AUC o 0.927 keia cohort, kikoo o 0.868, a me ka sensitivity o 0.820 no ka wanana ana i ka ino o na nodule ana 6 a 20 mm (Fig. 3e, f).Hōʻike kā mākou hopena e hiki i ka classifier ke hopu i nā loli metabolic i hoʻokumu ʻia e ka hoʻololi mua ʻana, me ka nānā ʻole i ka nui o ka nodule.
ad Ka hoʻohālikelike ʻana o nā moʻolelo PCA ma waena o nā pūʻulu i kuhikuhi ʻia e pili ana i kahi papa helu metabolic o 27 metabolites.CC a me BN <6 mm.b BN <6 mm vs BN 6–20 mm.ma LA 6–20 mm me LA 20–30 mm.g BN 6–20 mm a me 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 ʻO ka mea loaʻa ke ʻano hana (ROC) ka pihi e hōʻike ana i ka hana hoʻohālike hoʻokae no nā nodules 6-20 mm.f Ua helu ʻia nā waiwai kūpono ma muli o ke kumu hoʻohālike logistic regression no nā nodule e ana ana i 6-20 mm.Hōʻike ka laina kiko hina hina i ka waiwai ʻoki ʻoki maikaʻi loa (0.455).Hōʻike nā helu i luna i ka pākēneka o nā hihia i manaʻo ʻia no Los Angeles.E ho'ohana i ka ho'ā'o t a Haumāna huelo 'elua.ʻO ka PCA, ka nānāʻana i nā mea nui.AUC wahi ma lalo o ka pihi.Hāʻawi ʻia ka ʻikepili kumu ma ke ʻano o nā faila ʻikepili kumu.
ʻEhā mau hōʻailona (44-61 mau makahiki) me ka nui o ka pulmonary nodule like (7-9 mm) i koho hou ʻia e hōʻike i ka hana o ke kumu hoʻohālikelike i manaʻo ʻia (Fig. 4a, b).Ma ka nānā mua ʻana, hōʻike ʻia ka hihia 1 ma ke ʻano he nodule paʻa me ka calcification, kahi hiʻohiʻona e pili ana i ka maikaʻi, aʻo ka hihia 2 i hōʻike ʻia ma ke ʻano he nodule paʻa ʻole me ka ʻike ʻole o nā hiʻohiʻona maikaʻi.ʻEkolu mau pōʻai o ka hahai ʻana i ka CT scans i hōʻike ʻia ua kūpaʻa kēia mau hihia ma kahi o 4 mau makahiki a no laila ua manaʻo ʻia he nodules benign (Fig. 4a).Ke hoʻohālikelike ʻia me ka loiloi lapaʻau o ka serial CT scans, hoʻokahi-shot serum metabolite analysis me ka hoʻohālike classifier o kēia manawa me ka wikiwiki a ʻike pololei i kēia mau nodules benign e pili ana i nā palena probabilistic (Table 1).Hōʻike ka helu 4b i ka hihia 3 i kahi nodule me nā hōʻailona o ka hoʻihoʻi ʻana i ka pleural, i pili pinepine ʻia me ka malignancy32.ʻO ka hihia 4 i hōʻike ʻia ma ke ʻano he nodule paʻa paʻa ʻole me ka ʻike ʻole o kahi kumu maikaʻi.ʻO kēia mau hihia āpau i wānana ʻia ma ke ʻano he malignant e like me ke kumu hoʻohālike (Papa 1).Ua hōʻike ʻia ka loiloi o ka lung adenocarcinoma e ka histopathological examination ma hope o ka ʻokiʻoki ʻana o ka māmā (Fig. 4b).No ka hoʻonohonoho hōʻoia o waho, ua wānana pololei ka papa helu metabolic ʻelua mau hihia o nā nodules māmā indeterminate ʻoi aku ka nui ma mua o 6 mm (Supplementary Figure 10).
Nā kiʻi CT o ka puka makani axial o nā māmā o ʻelua mau hihia o nā nodules benign.Ma ka hihia 1, ua hōʻike ʻo CT scan ma hope o 4 mau makahiki i kahi nodule paʻa paʻa e ana ana i ka 7 mm me ka calcification ma ka ʻaoʻao lalo ʻākau.I ka hihia 2, ua hōʻike ʻia ka CT scan ma hope o 5 mau makahiki i kahi nodule paʻa paʻa paʻa me ke anawaena o 7 mm ma ka lobe luna ʻākau.b Axial window CT kiʻi o nā māmā a me nā noiʻi pathological e pili ana i ʻelua mau hihia o ka pae I adenocarcinoma ma mua o ka wehe ʻana i ka māmā.Hōʻike ka hihia 3 i kahi nodule me ke anawaena o 8 mm ma ka lobe luna ʻākau me ka hoʻihoʻi ʻana i ka pleural.Ua hōʻike ʻia ka hihia 4 i kahi nodule aniani lepo paʻa me 9 mm ma ka ʻaoʻao hema hema.Hematoxylin a me eosin (H&E) staining o resected lung tissue (scale bar = 50 μm) e hōʻike ana i ke ʻano ulu acinar o ka māmā adenocarcinoma.Hōʻike nā pua i nā nodule i ʻike ʻia ma nā kiʻi CT.ʻO nā kiʻi H&E he mau kiʻi hōʻikeʻike o nā māla microscopic he nui (>3) i nānā ʻia e ka mea maʻi.
Hoʻohui pū ʻia, hōʻike kā mākou mau hopena i ka waiwai kūpono o nā biomarkers metabolite serum i ka ʻike ʻokoʻa o nā nodules pulmonary, hiki ke hoʻohālikelike i ka wā e loiloi ana i ka nānā ʻana o CT.
Ma muli o kahi papa metabolite ʻokoʻa i hōʻoia ʻia, ʻimi mākou e ʻike i nā correlates biological o nā loli metabolic nui.ʻO ka nānāʻana i ka hoʻonuiʻana i ke ala KEGG e MetaboAnalyst iʻikeʻia he 6 mau ala i hoʻololi nuiʻia ma waena o nā huiʻelua (LA vs. HC a me LA vs. BN, hoʻololi p ≤ 0.001, hopena> 0.01).ʻO kēia mau hoʻololi i hōʻike ʻia e nā haunaele i ka pyruvate metabolism, tryptophan metabolism, niacin a me nicotinamide metabolism, glycolysis, TCA cycle, a me ka purine metabolism (Fig. 5a).A laila hana hou mākou i nā metabolomics i manaʻo ʻia e hōʻoia i nā loli nui me ka hoʻohana ʻana i ka helu piha.ʻO ka hoʻoholo ʻana i nā metabolites maʻamau i nā ala i hoʻololi pinepine ʻia e triple quadrupole mass spectrometry (QQQ) me ka hoʻohana ʻana i nā kūlana metabolite maoli.Hoʻokomo ʻia nā hiʻohiʻona demographic o ka metabolomics study target sample i ka Pākuʻi Pākuʻi 4. E like me kā mākou hopena metabolomics honua, ua hōʻoia ka quantitative analysis ua hoʻonui ʻia ka hypoxanthine a me ka xanthine, pyruvate, a me ka lactate ma LA i hoʻohālikelike ʻia me BN a me HC (Fig. 5b, c, p <0.05).Eia naʻe, ʻaʻohe ʻokoʻa koʻikoʻi o kēia mau metabolites i loaʻa ma waena o BN a me HC.
ʻO ka hoʻonui ʻana o ke ala KEGG i nā metabolites ʻokoʻa loa i ka hui LA i hoʻohālikelike ʻia i nā pūʻulu BN a me HC.Ua hoʻohana ʻia kahi Globaltest ʻelua-huelo, a ua hoʻoponopono ʻia nā helu p me ka hoʻohana ʻana i ke ʻano Holm-Bonferroni (hoʻoponopono ʻia p ≤ 0.001 a me ka nui o ka hopena > 0.01).b–d Nā ʻāpana Violin e hōʻike ana i ka hypoxanthine, xanthine, lactate, pyruvate, a me nā pae tryptophan i ka serum HC, BN, a me LA i hoʻoholo ʻia e LC-MS/MS (n = 70 no kēlā me kēia hui).Hōʻike nā laina kiko keʻokeʻo a me ka ʻeleʻele i ka median a me ka quartile.e Violin plot e hōʻike ana i ka Log2TPM maʻamau (transcripts per million) mRNA hōʻike o SLC7A5 a me QPRT i loko o ka adenocarcinoma māmā (n = 513) i hoʻohālikelike ʻia me ka ʻiʻo maʻamau maʻamau (n = 59) i ka ʻikepili LUAD-TCGA.ʻO ka pahu keʻokeʻo e hōʻike ana i ka laulā interquartile, ʻo ka laina ʻeleʻele ʻeleʻele ma ke kikowaena e hōʻike ana i ka median, a ʻo ka laina ʻeleʻele kūpaʻa e hoʻonui ana mai ka pahu e hōʻike ana i ka 95% confidence interval (CI).f Pearson correlation plot of SLC7A5 and GAPDH expression in lung adenocarcinoma (n = 513) a me ka maʻamau lung tissue (n = 59) i ka TCGA dataset.Hōʻike ka ʻāpana hina i ka 95% CI.r, ka huina hoopili Pearson.g Nā pae tryptophan cellular maʻamau i loko o nā pūnaewele A549 i hoʻololi ʻia me ka mana shRNA nonspecific (NC) a me shSLC7A5 (Sh1, Sh2) i hoʻoholo ʻia e LC-MS/MS.Hōʻike ʻia ka ʻikepili helu o ʻelima mau laʻana kūʻokoʻa biologically i kēlā me kēia hui.h Nā pae kelepona o NADt (ka huina NAD, me NAD+ a me NADH) i nā pūnaewele A549 (NC) a me SLC7A5 knockdown A549 cell (Sh1, Sh2).Hōʻike ʻia ka ʻikepili helu o nā laʻana kūʻokoʻa biologically i kēlā me kēia hui.i Ua ana ʻia ka hana Glycolytic o nā pūnaewele A549 ma mua a ma hope o ke kīkēkē ʻana o SLC7A5 e ka extracellular acidification rate (ECAR) (n = 4 biologically kūʻokoʻa hoʻohālike i kēlā me kēia hui).2-DG,2-deoxy-D-glucose.Ua hoʻohana ʻia ka hoʻāʻo t a ka Haumāna huelo ʻelua ma (b–h).Ma (g-i), hōʻike nā pahu kuhi i ka mean ± SD, ua hana ʻia kēlā me kēia hoʻokolohua ʻekolu manawa kūʻokoʻa a ua like nā hopena.Hāʻawi ʻia ka ʻikepili kumu ma ke ʻano o nā faila ʻikepili kumu.
Ke noʻonoʻo nei i ka hopena koʻikoʻi o ka metabolism tryptophan i hoʻololi ʻia i ka hui LA, ua loiloi pū mākou i nā pae tryptophan serum i nā pūʻulu HC, BN, a me LA me ka hoʻohana ʻana i QQQ.Ua ʻike mākou ua hoʻemi ʻia ka serum tryptophan ma LA i hoʻohālikelike ʻia me HC a i ʻole BN (p <0.001, Figure 5d), i kūlike me nā ʻike mua e ʻoi aku ka haʻahaʻa o ka pae tryptophan i nā poʻe maʻi me ka maʻi kanesa ma mua o nā mana olakino mai ka pūʻulu mana33,34 ,35.ʻO kekahi haʻawina e hoʻohana ana i ka PET/CT tracer 11C-methyl-L-tryptophan i ʻike ʻia ua hoʻonui nui ʻia ka manawa paʻa o ka tryptophan hōʻailona i loko o ka ʻiʻo maʻi maʻi maʻi maʻi i hoʻohālikelike ʻia i nā leona maikaʻi a i ʻole ka ʻiʻo maʻamau36.Manaʻo mākou ʻo ka emi ʻana o ka tryptophan i loko o ka serum LA e hōʻike ana i ka hana tryptophan ikaika e nā maʻi maʻi maʻi pūpū.
Ua ʻike ʻia ʻo ka hopena hope o ke ala kynurenine o ka tryptophan catabolism ʻo NAD + 37,38, he mea nui ia no ka hopena o ka glyceraldehyde-3-phosphate me 1,3-bisphosphoglycerate i glycolysis39.ʻOiai ua pili nā haʻawina mua i ka hana o ka tryptophan catabolism i ka hoʻoponopono immune, ua ʻimi mākou e wehewehe i ka interplay ma waena o ka tryptophan dysregulation a me nā ala glycolytic i ʻike ʻia i ka noiʻi o kēia manawa.Ua ʻike ʻia ʻo Solute transporter ʻohana 7 lālā 5 (SLC7A5) he tryptophan transporter43,44,45.ʻO ka quinolinic acid phosphoribosyltransferase (QPRT) he enzyme aia ma lalo o ke ala kynurenine e hoʻololi i ka quinolinic acid i NAMN46.ʻO ka nānāʻana i ka LUAD TCGA dataset i hōʻikeʻia ua hoʻonui nuiʻiaʻo SLC7A5 a me QPRT i loko o kaʻiʻo tumo i hoʻohālikelikeʻia me ke kino maʻamau (Fig. 5e).Ua ʻike ʻia kēia piʻi ʻana ma nā pae I a me II a me nā pae III a me IV o ka adenocarcinoma māmā (Supplementary Figure 11), e hōʻike ana i nā haunaele mua i ka tryptophan metabolism e pili ana me ka tumorigenesis.
Eia hou, ua hōʻike ka ʻikepili LUAD-TCGA i kahi pilina maikaʻi ma waena o SLC7A5 a me GAPDH mRNA hōʻike i nā maʻi maʻi maʻi maʻi (r = 0.45, p = 1.55E-26, Kiʻi 5f).I ka hoʻokaʻawale ʻana, ʻaʻohe pilina koʻikoʻi i ʻike ʻia ma waena o ia mau pūlima gene i loko o ka ʻiʻo māmā maʻamau (r = 0.25, p = 0.06, Kiʻi 5f).Knockdown o SLC7A5 (Supplementary Figure 12) ma A549 cell i hoʻemi nui i ka cellular tryptophan a me NAD (H) pae (Figure 5g, h), ka hopena o ka hana glycolytic attenuated e like me ke ana ʻia e ka extracellular acidification rate (ECAR) (Figure 1).5i).No laila, e pili ana i nā loli metabolic i ka serum a me ka ʻike in vitro, manaʻo mākou e hiki i ka tryptophan metabolism ke hana i ka NAD + ma o ke ala kynurenine a hana nui i ka hoʻolaha ʻana i ka glycolysis i ka maʻi kanesa.
Ua hōʻike nā haʻawina i ka nui o nā nodules pulmonary indeterminate i ʻike ʻia e LDCT hiki ke alakaʻi i ka pono no ka hoʻāʻo hou ʻana e like me PET-CT, biopsy lung, a me ka overtreatment ma muli o kahi hōʻailona maikaʻi ʻole o ka malignancy.31 E like me ka mea i hōʻike ʻia ma ka Figure 6, ʻIke ʻia kā mākou noiʻi ʻana i kahi papa o nā metabolites serum me ka waiwai diagnostic hiki ke hoʻomaikaʻi i ka stratification pilikia a me ka hoʻokele hope ʻana o nā nodules pulmonary i ʻike ʻia e CT.
Hoʻopili ʻia nā nodules pulmonary me ka hoʻohana ʻana i ka helu helu haʻahaʻa haʻahaʻa (LDCT) me nā hiʻohiʻona kiʻi e hōʻike ana i nā kumu maikaʻi a ʻino paha.ʻO ka hopena maopopo ʻole o nā nodules hiki ke alakaʻi i ka hele pinepine ʻana, nā hana pono ʻole, a me ka hoʻomaʻamaʻa.ʻO ka hoʻokomo ʻia ʻana o nā papa helu serum metabolic me ka waiwai diagnostic hiki ke hoʻomaikaʻi i ka loiloi pilikia a me ka mālama ʻana i nā nodules pulmonary.PET positron emission tomography.
Hōʻike ka ʻikepili mai ka US NLST study a me ka European NELSON study i ka nānā ʻana i nā hui kiʻekiʻe me ka haʻahaʻa haʻahaʻa computed tomography (LDCT) hiki ke hōʻemi i ka make maʻi maʻi maʻi lung1,3.Eia nō naʻe, ʻo ka loiloi pilikia a me ka hoʻokele hoʻokele ma hope o nā helu nui o nā nodules pulmonary incidental i ʻike ʻia e LDCT e mau ana ka paʻakikī loa.ʻO ka pahuhopu nui, ʻo ia ka hoʻopaʻa ʻana i ka hoʻokaʻawale pololei ʻana o nā protocols e pili ana i ka LDCT ma o ka hoʻokomo ʻana i nā biomarkers hilinaʻi.
Ua ʻike ʻia kekahi mau biomarkers molecular, e like me nā metabolites koko, ma ka hoʻohālikelike ʻana i ka maʻi maʻi maʻi me nā mana olakino15,17.Ma ka noiʻi o kēia manawa, ua kālele mākou i ka hoʻohana ʻana i ka serum metabolomics analysis e hoʻokaʻawale i waena o nā nodules pulmonary benign a malignant i ʻike ʻia e LDCT.Ua hoʻohālikelike mākou i ka honua serum metabolome o ka mana olakino (HC), benign lung nodules (BN), a me ka stage I lung adenocarcinoma (LA) samples me ka hoʻohana ʻana i ka nānā ʻana UPLC-HRMS.Ua ʻike mākou ua like ka HC a me BN i nā profile metabolic like, akā ua hōʻike ʻo LA i nā loli nui i hoʻohālikelike ʻia me HC a me BN.Ua ʻike mākou i ʻelua pūʻulu o nā metabolites serum e hoʻokaʻawale iā LA mai HC a me BN.
ʻO ka papahana ʻike LDCT i kēia manawa no nā nodules maikaʻi a me nā nodules ma muli o ka nui, density, morphology a me ka ulu ʻana o nā nodules i ka manawa30.Ua hōʻike ʻia nā haʻawina mua i ka nui o nā nodules e pili kokoke ana i ka hiki ke loaʻa i ka maʻi kanesa.ʻOiai i nā poʻe maʻi kiʻekiʻe, ʻo ka pilikia o ka maʻi maʻi ma nā nodes <6 mm he <1%.ʻO ka pilikia o ka maʻi maʻi no nā nodule e ana ana 6 a 20 mm mai 8% a 64%30.No laila, paipai ka Fleischner Society i ke anawaena oki o 6 mm no ka hahai mau CT.29 Eia naʻe, ʻaʻole i hoʻokō pono ʻia ka loiloi pilikia a me ka hoʻokele ʻana i nā nodules pulmonary indeterminate (IPN) ʻoi aku ka nui ma mua o 6 mm 31.ʻO ka mālama ʻana i ka maʻi naʻau congenital i kēia manawa e hoʻokumu ʻia ma ke kali makaʻala me ka nānā pinepine ʻana o CT.
Ma muli o ka metabolome i hōʻoia ʻia, ua hōʻike mākou no ka manawa mua i ka uhi ʻana o nā pūlima metabolomic ma waena o nā kānaka olakino a me nā nodules benign <6 mm.Ua kūlike ka like o ke olaola me nā ʻike CT mua e like me ka haʻahaʻa o ka maʻi ʻino no nā nodules <6 mm e like me nā kumuhana me ka ʻole o nā nodes.30 Pono e hoʻomaopopo ʻia e hōʻike pū ana kā mākou hopena i nā nodules benign <6 mm a me ≥6 mm he kiʻekiʻe. like i loko o ka metabolomic profiles, e hōʻike ana i ka wehewehe hana o ka benign etiology e kūlike me ka nānā ʻole i ka nui o ka nodule.No laila, hiki i nā panela serum metabolite diagnostic hou ke hāʻawi i hoʻokahi assay ma ke ʻano he hoʻāʻo ʻana i ka wā i ʻike mua ʻia nā nodules ma CT a hiki ke hōʻemi i ka nānā ʻana.I ka manawa like, ua ʻike ka papa like o nā biomarkers metabolic i nā nodules malignant ≥6 mm i ka nui mai nā nodules benign a hāʻawi i nā wānana pololei no nā IPN o ka nui like a me nā hiʻohiʻona morphological ambiguous ma nā kiʻi CT.Ua hana maikaʻi kēia mea hoʻohālikelike metabolism serum i ka wānana ʻana i ka maʻi o nā nodules ≥6 mm me kahi AUC o 0.927.Hoʻohui pū ʻia, hōʻike kā mākou mau hopena i nā hōʻailona serum metabolomic kūʻokoʻa e hōʻike pono i nā loli metabolic i hoʻokomo ʻia i ka maʻi maʻi a loaʻa ka waiwai kūpono e like me nā mea wānana pilikia, kūʻokoʻa i ka nui o ka nodule.
ʻO ka mea nui, ʻo ka lung adenocarcinoma (LUAD) a me ka squamous cell carcinoma (LUSC) nā ʻano nui o ka maʻi maʻi ʻaʻai liʻiliʻi liʻiliʻi (NSCLC).Ma muli o ka pili nui ʻana o LUSC me ka hoʻohana ʻana i ka paka47 a ʻo LUAD ka mōʻaukala maʻamau o nā nodules māmā i ʻike ʻia ma CT screening48, ua kūkulu pono ʻia kā mākou kumu hoʻohālike no ka pae I adenocarcinoma samples.Ua nānā pū ʻo Wang a me nā hoa hana iā LUAD a ʻike i ʻeiwa mau pūlima lipid e hoʻohana ana i ka lipidomics e hoʻokaʻawale i ka maʻi maʻi maʻi maʻi mua o ka poʻe olakino17.Ua hoʻāʻo mākou i ke kumu hoʻohālikelike o kēia manawa ma 16 mau hihia o ka pae I LUSC a me 74 benign nodules a ʻike i ka haʻahaʻa haʻahaʻa wānana LUSC (AUC 0.776), e hōʻike ana e loaʻa paha iā LUAD a me LUSC kā lākou mau inoa metabolomic ponoʻī.ʻOiaʻiʻo, ua hōʻike ʻia ʻo LUAD a me LUSC he ʻokoʻa i ka etiology, ke kumu ola a me nā aberrations genetic49.No laila, pono e hoʻokomo ʻia nā ʻano histology ʻē aʻe i loko o nā kumu hoʻohālike no ka ʻike ʻana i ka maʻi maʻi maʻi maʻi maʻi ma nā papahana screening.
Maʻaneʻi, ua ʻike mākou i nā ala ala ʻeono i hoʻololi pinepine ʻia i ka adenocarcinoma māmā i hoʻohālikelike ʻia me nā mana olakino a me nā nodules maikaʻi.ʻO ka Xanthine a me ka hypoxanthine nā metabolites maʻamau o ka purine metabolic pathway.E like me kā mākou hopena, ua hoʻonui nui ʻia nā mea waena e pili ana me ka purine metabolism i ka serum a i ʻole nā kikoʻī o nā maʻi me ka adenocarcinoma māmā i hoʻohālikelike ʻia me nā mana olakino a i ʻole nā mea maʻi ma ka pae preinvasive15,50.Hiki ke hōʻike ʻia nā pae kiʻekiʻe o ka serum xanthine a me ka hypoxanthine i ka anabolism e koi ʻia e ka hoʻonui wikiwiki ʻana i nā maʻi kanesa.ʻO ka dysregulation o ka glucose metabolism kahi hōʻailona kaulana o ka maʻi kanesa metabolism51.Maʻaneʻi, uaʻike mākou i ka piʻi nuiʻana o ka pyruvate a me ka lactate i ka hui LA i hoʻohālikelikeʻia me ka hui HC a me BN, i kūlike me nā hōʻike mua o nā meaʻinoʻole o ke ala glycolytic i loko o ka serum metabolome profiles o nā maʻi maʻi maʻi maʻiʻole (NSCLC) a nā mana olakino.ua kūlike nā hopena52,53.
ʻO ka mea nui, ua ʻike mākou i ka hoʻopili ʻana ma waena o ka pyruvate a me ka tryptophan metabolism i ka serum o nā adenocarcinomas lung.Ua hōʻemi ʻia nā pae serum tryptophan i ka hui LA i hoʻohālikelike ʻia me ka hui HC a i ʻole BN.ʻO ka mea e mahalo ai, ua ʻike ʻia kahi haʻawina nui ma mua e hoʻohana ana i kahi cohort prospective e pili ana nā haʻahaʻa haʻahaʻa o ka tryptophan e holo nei me ka piʻi nui ʻana o ka maʻi kanesa 54.ʻO Tryptophan kahi waikawa amino koʻikoʻi i loaʻa iā mākou mai ka meaʻai.Manaʻo mākou ʻo ka pau ʻana o ka tryptophan serum i ka adenocarcinoma māmā e hōʻike ana i ka hoʻopau wikiwiki ʻana o kēia metabolite.Ua ʻike ʻia ʻo ka hopena hope o ka tryptophan catabolism ma o ke ala kynurenine ke kumu o ka de novo NAD + synthesis.No ka mea ua hana mua ʻia ʻo NAD + ma o ke ala hoʻopakele, ʻo ke koʻikoʻi o NAD + i ka tryptophan metabolism i ke olakino a me nā maʻi e hoʻoholo ʻia46.ʻO kā mākou hōʻike ʻana i ka waihona TCGA ua hōʻike ʻia ua hoʻonui nui ʻia ka ʻōlelo o ka tryptophan transporter solute transporter 7A5 (SLC7A5) i loko o ka adenocarcinoma māmā i hoʻohālikelike ʻia me nā mana maʻamau a ua hoʻopili maikaʻi ʻia me ka hōʻike o ka glycolytic enzyme GAPDH.ʻO nā haʻawina mua i kālele nui ʻia i ka hana o ka tryptophan catabolism i ke kāohi ʻana i ka pane antitumor immune40,41,42.Ma aneʻi mākou e hōʻike nei i ka pale ʻana i ka tryptophan uptake e ka knockdown o SLC7A5 i loko o nā maʻi maʻi maʻi pūpū e hopena i ka emi ʻana o nā pae NAD cellular a me kahi attenuation concomitant o ka hana glycolytic.I ka hōʻuluʻulu ʻana, hāʻawi kā mākou noiʻi i kahi kumu ola no nā loli i ka metabolism serum e pili ana me ka hoʻololi ʻino ʻana o ka adenocarcinoma māmā.
ʻO nā hoʻololi EGFR ka hoʻololi maʻamau o ka mea hoʻokele i nā maʻi me NSCLC.Ma kā mākou noiʻi ʻana, ʻike mākou ua loaʻa nā maʻi me ka mutation EGFR (n = 41) i nā ʻano metabolomic holoʻokoʻa e like me nā maʻi me ka EGFR ʻano ʻāhiu (n = 31), ʻoiai ua ʻike mākou i ka emi ʻana o nā pae serum o kekahi mau maʻi mutant EGFR i nā maʻi acylcarnitine.ʻO ka hana i hoʻokumuʻia o acylcarnitines,ʻo ia ka laweʻana i nā hui acyl mai ka cytoplasm i loko o ka matrix mitochondrial, e alakaʻi ana i ka oxidation o nā momona momona e hana i ka ikehu 55.E like me kā mākou ʻike, ua ʻike ʻia kahi noiʻi hou i nā ʻano metabolome like ma waena o EGFR mutant a me EGFR wild-type tumors ma ka nānā ʻana i ka metabolome honua o 102 lung adenocarcinoma tissue samples50.ʻO ka mea mahalo, ua loaʻa pū ka ʻike acylcarnitine i ka hui mutant EGFR.No laila, inā paha e hōʻike ana nā hoʻololi i nā pae acylcarnitine i nā hoʻololi metabolic i hoʻokomo ʻia e EGFR a me nā ala molekala lalo e pono ke aʻo hou.
I ka hopena, hoʻokumu kā mākou noiʻi ʻana i kahi papa helu serum metabolic no ka maʻi ʻokoʻa o nā nodules pulmonary a hoʻolālā i kahi kaʻina hana e hiki ke koho i ka loiloi pilikia a hoʻomaʻamaʻa i ka hoʻokele lapaʻau e pili ana i ka nānā ʻana i ka CT scan.
Ua ʻae ʻia kēia haʻawina e ke Kōmike Ethics o Sun Yat-sen University Cancer Hospital, ka First Affiliated Hospital o Sun Yat-sen University, a me ke Kōmike Ethics o Zhengzhou University Cancer Hospital.Ma ka ʻike a me nā pūʻulu hōʻoia kūloko, ua hōʻiliʻili ʻia ka 174 sera mai nā poʻe olakino a me 244 sera mai nā nodules benign mai nā poʻe e hana ana i nā hoʻokolohua olakino makahiki ma ke Keʻena ʻo Cancer Control and Prevention, Sun Yat-sen University Cancer Center, a me 166 benign nodules.serum.Ua hōʻiliʻili ʻia nā adenocarcinoma ma ka pae I mai Sun Yat-sen University Cancer Center.I loko o ka hui hōʻoia o waho, aia he 48 mau hihia o nā nodules maikaʻi, 39 mau hihia o ka pae I lung adenocarcinoma mai ka Halemai Hoʻohui Mua o Sun Yat-sen University, a me 24 mau hihia o ka pae I lung adenocarcinoma mai Zhengzhou Cancer Hospital.Ua hōʻiliʻili pū ʻo Sun Yat-sen University Cancer Center i 16 mau hihia o ka pae I squamous cell lung cancer no ka hoʻāʻo ʻana i ka hiki ke diagnostic o ka papa helu metabolic i hoʻokumu ʻia (hōʻike ʻia nā ʻano maʻi ma ka Pākuʻi 5).Ua hōʻiliʻili ʻia nā laʻana mai ka cohort ʻike a me ka cohort hōʻoia kūloko ma waena o Ianuali 2018 a me Mei 2020. Ua hōʻiliʻili ʻia nā laʻana no ka hui hōʻoia waho ma waena o ʻAukake 2021 a me ʻOkakopa 2022. pūʻulu.Hui ʻIke a me ka Pūʻulu Nānā Kuloko.Hoʻoholo ʻia ke kāne komo ma muli o ka hōʻike pilikino.Ua loaʻa ka ʻae ʻike mai nā mea komo āpau a ʻaʻohe uku i hāʻawi ʻia.ʻO nā kumuhana me nā nodules maikaʻi ka poʻe me ka helu CT scan paʻa ma 2 a 5 mau makahiki i ka manawa o ka nānā ʻana, koe wale nō ka hihia 1 mai ka hōʻoia hōʻoia waho, i hōʻiliʻili ʻia ma mua o ka hana a ʻike ʻia e ka histopathology.Koe nae ka bronchitis mau.Ua hōʻiliʻili ʻia nā maʻi adenocarcinoma ma mua o ka wehe ʻana o ka māmā a hōʻoia ʻia e ka maʻi pathological.Ua hōʻiliʻili ʻia nā laʻana koko hoʻokēʻai i nā paipu hoʻokaʻawale serum me ka ʻole o nā anticoagulants.Hoʻopili ʻia nā koko koko no 1 hola ma ka lumi wela a laila centrifuged ma 2851 × g no 10 mau minuke ma 4 ° C e hōʻiliʻili i ka serum supernatant.Ua hoʻopaʻa ʻia nā serum aliquots ma -80 ° C a hiki i ka lawe ʻana i ka metabolite.Ua hōʻiliʻili ke Keʻena ʻOihana Kāohi a me ka Nānā Lapaʻau o ka Sun Yat-sen University Cancer Center i kahi wai o ka serum mai 100 mau mea hāʻawi olakino, me ka helu like o nā kāne a me nā wahine mai 40 a 55 mau makahiki.Ua hui pū ʻia ka nui like o kēlā me kēia hāpana hāʻawi, ua hoʻokaʻawale ʻia ka ʻauʻau a mālama ʻia ma -80°C.Ua hoʻohana ʻia ka hui ʻana o ka serum ma ke ʻano he mea kuhikuhi no ka mālama ʻana i ka maikaʻi a me ka standardization data.
Ua hoʻoheheʻe ʻia ka serum kuhikuhi a me nā laʻana hoʻāʻo a lawe ʻia nā metabolites me ka hoʻohana ʻana i kahi ala hoʻohui hui (MTBE/methanol/wai) 56.ʻO ka pōkole, ua hui pū ʻia ka 50 μl o ka serum me 225 μl o ka methanol hau anuanu a me 750 μl o ka methyl tert-butyl ether (MTBE).E hoʻoulu i ka hui a hoʻokomo i ka hau no 1 hola.A laila hui ʻia nā laʻana a hui pū ʻia ka vortex me 188 μl o ka wai MS-grade i loaʻa nā kūlana kūloko (13C-lactate, 13C3-pyruvate, 13C-methionine, a me 13C6-isoleucine, i kūʻai ʻia mai Cambridge Isotope Laboratories).A laila ua centrifuged ka hui ʻana ma 15,000 × g no 10 min ma 4 ° C, a ua hoʻololi ʻia ka pae haʻahaʻa i ʻelua mau paipu (125 μL kēlā me kēia) no ka nānā ʻana o LC-MS ma nā ʻano maikaʻi a maikaʻi ʻole.ʻO ka hope, ua hoʻoheheʻe ʻia ka hāpana a maloʻo i loko o kahi concentrator māmā holo wikiwiki.
Hoʻohui hou ʻia nā metabolites maloʻo i 120 μl o 80% acetonitrile, vortexed no 5 min, a centrifuged ma 15,000 × g no 10 min ma 4 ° C.Ua hoʻoili ʻia nā supernatants i loko o nā hue aniani amber me nā microinserts no nā haʻawina metabolomics.Ka nānā ʻana i ka metabolomics ʻaʻole i hoʻopaʻa ʻia ma kahi ʻano ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) platform.Ua hoʻokaʻawale ʻia nā metabolites me ka ʻōnaehana Dionex Ultimate 3000 UPLC a me kahi kolamu ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Wai).Ma ke ʻano ion maikaʻi, ʻo 95% (A) a me 50% acetonitrile (B) nā pae kelepona, ʻo kēlā me kēia me 10 mmol/L ammonium acetate a me 0.1% formic acid.Ma keʻano maikaʻiʻole, loaʻa ka 95% a me ka 50% acetonitrile i nā māhele kelepona A me B, i kēlā me kēia, aia nā māheleʻelua i ka 10 mmol / L ammonium acetate, pH = 9. ʻO ka papahana gradient penei: 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. Ua mālama ʻia ke kolamu ma 40 ° C a me ka hāpana ma 10 ° C i ka autosampler.ʻO 0.3 ml / min ka kahe kahe, ʻo ka nui o ka injection he 3 μl.Ua hoʻohana ʻia kahi Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific) me kahi kumu electrospray ionization (ESI) i ke ʻano scan piha a hui pū ʻia me ke ʻano kiaʻi ddMS2 e hōʻiliʻili i ka nui o ka ʻikepili.Ua hoʻonohonoho ʻia nā ʻāpana MS penei: spray voltage +3.8 kV/- 3.2 kV, capillary temperature 320°C, shielding gas 40 arb, auxiliary gas 10 arb, probe heat heat 350°C, scanning range 70–1050 m/h, olelo hooholo.70 000. Ua loaʻa ka ʻikepili me Xcalibur 4.1 (Thermo Fisher Scientific).
No ka loiloi ʻana i ka maikaʻi o ka ʻikepili, ua hoʻokumu ʻia nā mea hoʻohālike i hoʻohui ʻia i ka mana maikaʻi (QC) ma ka wehe ʻana i 10 μL aliquots o ka supernatant mai kēlā me kēia hāpana.Ua kālailai ʻia ʻeono mau ʻokoʻa hōʻailona mana maikaʻi ma ka hoʻomaka ʻana o ke kaʻina analytical e nānā i ka paʻa o ka ʻōnaehana UPLC-MS.Hoʻokomo ʻia nā laʻana hoʻomalu maikaʻi i kēlā me kēia manawa i ka pūʻulu.ʻO nā pūʻulu 11 a pau o nā laʻana serum i loko o kēia haʻawina i nānā ʻia e LC-MS.Ua hoʻohana ʻia nā Aliquots o kahi hui wai serum mai 100 mau mea hāʻawi olakino ma ke ʻano he kumu kuhikuhi i kēlā me kēia pūʻulu e nānā i ke kaʻina hana ʻoki a hoʻoponopono no nā hopena batch-to-batch.ʻO ka loiloi metabolomics untargeted o ka cohort ʻike, ka cohort validation i loko, a me ka cohort hōʻoia waho i hana ʻia ma ka Metabolomics Center o ke Kulanui o Sun Yat-sen.ʻO ka hale hana waho o Guangdong University of Technology Analysis and Testing Center i kālailai ʻia he 40 mau laʻana mai ka hui waho e hoʻāʻo i ka hana o ke kumu hoʻohālike.
Ma hope o ka unuhi ʻana a me ka hoʻoponopono hou ʻana, ua ana ʻia ka nui piha o nā metabolites serum me ka hoʻohana ʻana i ka ultra-high performance liquid chromatography-tandem mass spectrometry (Agilent 6495 triple quadrupole) me kahi kumu electrospray ionization (ESI) i ke ʻano he nui o ka nānā ʻana i ka hopena (MRM).Ua hoʻohana ʻia kahi kolamu ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Wai) e hoʻokaʻawale i nā metabolites.ʻO ka māhele kelepona he 90% (A) a me 5% acetonitrile (B) me 10 mmol / L ammonium acetate a me 0.1% ammonia solution.Penei ka papahana gradient: 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.Ua mālama ʻia ke kolamu ma 40 °C a me ka hāpana ma 10 °C i ka autosampler.ʻO 0.3 mL / min ka kahe kahe a ʻo ka nui o ka injection he 1 μL.Ua hoʻonohonoho ʻia nā ʻāpana MS penei: capillary voltage ± 3.5 kV, nebulizer pressure 35 psi, sheath gas kahe 12 L / min, sheath gas wela 350 ° C, maloʻo kinoea wela 250 ° C, a maloʻo kinoea kahe 14 l / min.ʻO nā hoʻololi MRM o tryptophan, pyruvate, lactate, hypoxanthine a me xanthine he 205.0–187.9, 87.0–43.4, 89.0–43.3, 135.0–92.3 a me 151.0–107.9 pakahi.Ua hōʻiliʻili ʻia ka ʻikepili me ka hoʻohana ʻana iā Mass Hunter B.07.00 (Agilent Technologies).No nā laʻana serum, ua helu ʻia ka tryptophan, pyruvate, lactate, hypoxanthine, a me xanthine me ka hoʻohana ʻana i nā pihi calibration o nā hoʻonā hui ʻana maʻamau.No nā laʻana kelepona, ua hoʻohālikelike ʻia ka ʻike tryptophan i ka maʻamau o loko a me ka nui o ka protein cell.
Ua hoʻohana ʻia ka hoʻohana ʻana i ka Compound Discovery 3.1 a me TraceFinder 4.0 (Thermo Fisher Scientific).No ka hoʻopau ʻana i nā ʻokoʻa like ʻole ma waena o nā pūʻulu, ua puʻunaue ʻia kēlā me kēia peak hiʻohiʻona o ka hāpana hoʻāʻo e ka piko hiʻona o ka mea kuhikuhi mai ka pūʻulu like e loaʻa ai ka nui pili.Hōʻike ʻia nā ʻokoʻa ma waena o nā pūʻulu ʻelua e ka helu ʻike wahaheʻe (FDR<0.05, Wilcoxon hōʻoia kūlana) a me ka hoʻololi ʻana (>1.2 a i ʻole <0.83).Hōʻike ʻia ka ʻikepili MS Raw o nā hiʻohiʻona i unuhi ʻia a me ka ʻikepili MS i hoʻoponopono ʻia i ka serum i hōʻike ʻia ma ka ʻikepili Hoʻohui 1 a me ka ʻikepili Hoʻohui 2, kēlā me kēia.Ua hana ʻia ka hōʻike kiʻekiʻe ma muli o nā pae i wehewehe ʻia o ka ʻike, me nā metabolites i ʻike ʻia, nā pūhui i hoʻopaʻa ʻia i hoʻopaʻa ʻia, nā papa pūhui i hōʻike ʻia, a me nā pūhui ʻike ʻole 22.Ma muli o ka ʻimi ʻikepili ma Compound Discovery 3.1 (mzCloud, HMDB, Chemspider), ua koho hope ʻia nā pūhui olaola me MS/MS kūpono i nā kūlana i hoʻopaʻa ʻia a i ʻole nā hōʻike hoʻohālikelike pololei ma mzCloud (helu > 85) a i ʻole Chemspider i koho hope ʻia ma waena o ka metabolome ʻokoʻa.Hoʻokomo ʻia nā hōʻike kiʻekiʻe no kēlā me kēia hiʻohiʻona i ka ʻikepili hou 3. Ua hoʻohana ʻia ka MetaboAnalyst 5.0 no ka loiloi univariate o ka nui o ka metabolite maʻamau.Ua loiloi pū ʻo MetaboAnalyst 5.0 i ka nānā ʻana i ka hoʻonui ʻana i ke ala KEGG ma muli o nā metabolites ʻokoʻa loa.Ua kālailai ʻia ka nānā ʻana i nā ʻāpana kumu (PCA) a me ka nānā ʻana i nā ʻāpana liʻiliʻi liʻiliʻi loa (PLS-DA) me ka hoʻohana ʻana i ka pūʻolo polokalamu ropls (v.1.26.4) me ka stack normalization a me ka autoscaling.Ua hana ʻia ke kumu hoʻohālike biomarker metabolite maikaʻi loa no ka wānana ʻana i ka maʻi nodule me ka hoʻohana ʻana i ka regression logistic binary me ka liʻiliʻi loa a me ka mea koho (LASSO, R package v.4.1-3).Ua hōʻike ʻia ka hana ʻana o ka hoʻohālike hoʻokaʻawale i ka ʻike a me ka hōʻoia ʻana ma ka helu ʻana i ka AUC ma muli o ka loiloi ROC e like me ka pROC package (v.1.18.0.).Ua loaʻa ka ʻoki kūpono kūpono ma muli o ka ʻoi loa ʻo Youden index o ke kumu hoʻohālike (sensitivity + specificity - 1).ʻO nā laʻana me nā waiwai liʻiliʻi a ʻoi aku paha ma mua o ka paepae e wānana ʻia ʻo ia nā nodules maikaʻi a me ka adenocarcinoma māmā, kēlā me kēia.
Ua hoʻoulu ʻia nā pūnaewele A549 (#CCL-185, American Type Culture Collection) i loko o ke kikowaena F-12K me 10% FBS.Ua hoʻokomo ʻia nā ʻāpana lauoho pōkole RNA (shRNA) e kuhikuhi ana iā SLC7A5 a me kahi mana nontargeting (NC) i loko o ka lentiviral vector pLKO.1-puro.ʻO nā kaʻina antisense o shSLC7A5 penei: Sh1 (5′-GGAGAAACCTGATGAACAGTT-3′), Sh2 (5′-GCCGTGGACTTCGGGAACTAT-3′).Ua kūʻai ʻia nā antibody i ka SLC7A5 (#5347) a me ka tubulin (#2148) mai Cell Signaling Technology.Ua hoʻohana ʻia nā antibodies i ka SLC7A5 a me ka tubulin ma kahi dilution o 1: 1000 no ka nānā ʻana o Western blot.
ʻO ka Seahorse XF Glycolytic Stress Test e ana i nā pae hoʻokaʻina extracellular (ECAR).I ka ho'āʻo, glucose, oligomycin A, a me 2-DG i lawelawe ʻia e hoʻāʻo i ka mana glycolytic cellular e like me ke ana ʻana e ECAR.
Ua hoʻopili ʻia nā pūnaewele A549 me ka mana non-targeting control (NC) a me shSLC7A5 (Sh1, Sh2) i ka pō i loko o nā kīʻaha 10 cm anawaena.Ua unuhi ʻia nā metabolites pūnaewele me 1 ml o ka methanol wai anuanu 80%.Ua ʻoki ʻia nā cell i loko o ka solution methanol, hōʻiliʻili ʻia i loko o kahi paipu hou, a centrifuged ma 15,000 × g no 15 min ma 4°C.E hōʻiliʻili i 800 µl o ka supernatant a maloʻo me ka hoʻohana ʻana i kahi concentrator māmā holo wikiwiki.Hoʻopili ʻia nā pellets metabolite maloʻo no nā pae tryptophan e hoʻohana ana i ka LC-MS/MS e like me ka mea i hōʻike ʻia ma luna.Ua ana ʻia nā pae kelepona NAD (H) i nā pūnaewele A549 (NC a me shSLC7A5) me ka hoʻohana ʻana i kahi kit colorimetric NAD +/NADH (#K337, BioVision) e like me nā ʻōlelo a ka mea hana.Ua ana ʻia nā pae protein no kēlā me kēia laʻana e hoʻomaʻamaʻa i ka nui o nā metabolites.
ʻAʻole i hoʻohana ʻia nā ʻano helu helu no ka hoʻoholo mua ʻana i ka nui hāpana.ʻO nā haʻawina metabolomics mua e pili ana i ka biomarker discovery15,18 ua manaʻo ʻia he mau hōʻailona no ka hoʻoholo ʻana i ka nui, a hoʻohālikelike ʻia i kēia mau hōʻike, ua lawa kā mākou laʻana.ʻAʻohe laʻana i hoʻokaʻawale ʻia mai ka pūʻulu haʻawina.Ua hāʻawi wale ʻia nā ʻāpana serum i kahi hui ʻike (306 mau hihia, 74.6%) a me kahi hui hōʻoia kūloko (104 mau hihia, 25.4%) no nā haʻawina metabolomics ʻole.Ua koho maʻalahi mākou i nā hihia 70 mai kēlā me kēia hui mai ka ʻike i hoʻonohonoho ʻia no nā haʻawina metabolomics.Ua makapō ka poʻe noiʻi i ka hana pūʻulu i ka wā o ka hōʻiliʻili a me ka nānā ʻana o ka ʻikepili LC-MS.Hōʻike ʻia nā ʻikepili helu o ka ʻikepili metabolomics a me nā hoʻokolohua cell ma nā ʻāpana o nā hopena, nā moʻolelo kiʻi, a me nā ʻano.ʻO ka helu ʻana o ka tryptophan cellular, NADT, a me ka hana glycolytic i hana ʻia ʻekolu manawa kūʻokoʻa me nā hopena like.
No ka ʻike hou aku e pili ana i ka hoʻolālā haʻawina, e ʻike i ka Natural Portfolio Report Abstract pili me kēia ʻatikala.
Hōʻike ʻia ka ʻikepili MS maka o nā hiʻohiʻona i unuhi ʻia a me ka ʻikepili MS maʻamau o ka serum reference i hōʻike ʻia i ka ʻikepili Hoʻohui 1 a me ka ʻikepili Hoʻohui 2, kēlā me kēia.Hōʻike ʻia nā hōʻike kiʻekiʻe no nā hiʻohiʻona ʻokoʻa ma ka ʻikepili hou 3. Hiki ke hoʻoiho ʻia ka ʻikepili LUAD TCGA mai https://portal.gdc.cancer.gov/.Hāʻawi ʻia ka ʻikepili hoʻokomo no ka hoʻolālā ʻana i ka pakuhi ma ka ʻikepili kumu.Hāʻawi ʻia ka ʻikepili kumu no kēia ʻatikala.
National Lung Screening Study Group, etc. E hoemi ana i ka make ma'i ma'i ma'i me ka helu ha'aha'a.ʻEnelani ʻĀkau.J. Med.365, 395–409 (2011).
Kramer, BS, Berg, KD, Aberle, DR and Prophet, PC Lung cancer screening using low-dose helical CT: nā hualoaʻa mai ka National Lung Screening Study (NLST).J. Med.Papa 18, 109–111 (2011).
De Koning, HJ, et al.Ho'emi i ka make 'ana o ka ma'i ma'i 'a'ai me ka nānā 'ana i ka volumetric CT ma kahi ho'ā'o randomized.ʻEnelani ʻĀkau.J. Med.382, 503–513 (2020).
Ka manawa hoʻouna: Sep-18-2023