Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD073278/checksum.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD073278/XB05936DA_ctrl20_E2_1_32479.d.zipftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD073278/search.zipftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD073278/XB05936DA_cre21_E3_1_32481.d.zipprimaryOK200zhuhongrui@mail.ustc.edu.cnli wangMass SpectrometryBottom-up proteomicsMouseGlioblastomahttps://www.ebi.ac.uk/pride/archive/projects/PXD073278BrainAst Cre-On- receiver mice loaded with GL261 sender glioma cells and Astrocyte specific secretion protein mice (Aldh1l1-CreERT; Rosa26 TurboIDV5) loaded with GL261 glioma were treated subcutaneously with 500 μL of a 2-mg/mL biotin solution (Beyotime) once a day for five consecutive days. The tumor tissues were lysed in 1 ml urea lysis buffer (8 M urea, 10 mM Tris, 100 mM NaH2PO4, pH 8.5) containing protease and phosphatase inhibitors, without EDTA. Then, lysates were lysed with 30% ultrasound for 3 min (amplitude 30 W, switch pulse for 5 seconds), shaken at 4℃ for 30 min, and centrifuged at 12000 rpm for 10 min to obtain a supernatant rich in biotinylated proteins. At least 50 μg of the supernatant should be used for western blot before biotin beads enrichment. The remaining supernatant was added with 80~100 μL RIPA pre-washed Biotinylation Beads (Beyotime) and incubated at 4℃ overnight. The next day, the sample was centrifuged at 1200 rpm for 2 min to obtain beads rich in biotin protein. Then, beads were orderly washed on ice with 1 ml RIPA lysis buffer (containing protease phosphatase inhibitor) for a total of 5 min, with 1 ml of 1 M KCl for 8 min, rapidly washed in 1 ml of 0.1 M Na2CO3, 1 ml of 2 M urea [prepared with 10 mM Tris HCl (pH 8.0)] and twice with 1 ml of RIPA lysis buffer for 5 min. After steps completed, the biotin protein beads were obtained, and 20 μl of beads was took for western blot with 4 to 12% polyacrylamide gel or silver staining detection. The remaining beads were used for subsequent protein mass spectrometry identification.PrideNot availableThe samples were added appropriate amount of TEAB to adjust pH at 8.0. 5 μL of suspension was used for SDS-PAGE testing. And remaining samples were performed for digestion. The protein solution was reduced with 5 mM dithiothreitol for 30 min at 56℃ and alkylated with 11 mM iodoacetamide for 15 min at RT in darkness. The samples were then diluted by adding 200 mM TEAB to urea concentration less than 2 M. Next, trypsin (1:50 at trypsin-to-protein mass ratio) was added for the first digestion overnight and 1:100 for a second 4 h - digestion. Finally, the peptides were desalted by Strata X SPE column and separated on avanquish neo UPLC system (Thermo Fisher Scientific). The separated peptides were analyzed in Orbitrap Exploris 480 with a nano-electrospray ion source. Precursors and fragments were analyzed at the Orbitrap detector. The full MS scan resolution was set to 60000 for a scan range of 350-1400 m/z. The MS/MS scan was fixed first mass as 120.0 m/z at a resolution of 15000. The HCD fragmentation was performed at a normalized collision energy of 27%. Automatic gain control target was set at 1E6, with a maximum injection time of 22 ms. The DIA data were processed using DIA-NN search engine. Tandem mass spectra were searched against Mus_musculus_10090_SP_20231220.fasta concatenated with reverse decoy database. Trypsin/P was specified as cleavage enzyme allowing up to 1 missing cleavage. Excision on N-term Met and carbamidomethyl on Cys were specified as fixed modification. FDR was adjusted to < 1%. Gene ontology analysis, KEGG Pathway annotation, subcellular localization annotation, and protein-protein interaction network were performed for data analysis.ProteomicsHongrui ZhuPARTIALFirst Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of ChinaMus Musculus (mouse)Not availablewli1@mail.ustc.edu.cnCancer (b/d-hpp)Human proteome projectBiology/disease-driven human proteome project (b/d-hpp)University of Science and Technology of ChinaChinafalseMetabolic licensing of quiescent glioblastoma activation and immune evasion via astrocyte-mitochondria shuttleThe reaction of quiescent tumor cell subpopulations at the invasive margin of glioblastoma (GBM) might be cleared by immune surveillance toward death or expand through immune escape, which can drive therapeutic resistance, tumor dissemination, and recurrence through phenotypic plasticity. However, the process and mechanisms underlying their activation remain largely unclear. This study revealed that astrocytes awaken quiescent GBM cells to cycling and immune-evasive phenotypes. Astrocytic mitochondria are indispensable in regulating the phenotypic changes. Specifically, the SHMT2 enzyme in astrocytic mitochondria catalyzes one-carbon metabolic products, which significantly promote synthetic metabolic activation and reactivation of quiescent GBM cells to immune evasion through intercellular mitochondrial transfer. Tumor cells that acquire astrocytic mitochondria enhanced one-carbon metabolism drives global hyperactive anabolic metabolism through YTHDC1-mediated transcriptional pause release of ribosomal and focal adhesion signaling genes, supporting rapid proliferation and conferring a potent immune-evasive phenotype. Therapeutically, pharmacologic or genetic inhibiting the astrocytic SHMT2 or blocking mitochondrial transfer markedly suppressed quiescent GBM cell activation and tumor progression. These findings elucidate a novel mechanism by which astrocytes in the tumor microenvironment drive GBM malignancy through metabolic symbiosis.2026-07-302026-01-20PXD073278NEWT:6945NEWT:184922NEWT:6703NEWT:3555NEWT:71647NEWT:2NEWT:157546NEWT:474186NEWT:35554NEWT:38942NEWT:307972NEWT:32046NEWT:1496NEWT:544496NEWT:2102NEWT:2042546NEWT:259447NEWT:45351NEWT:445974NEWT:43179NEWT:4513NEWT:180454NEWT:5722NEWT:1247NEWT:6938NEWT:1129NEWT:376741NEWT:55153NCBITaxon:10407NEWT:1736309NEWT:309800NEWT:281395NEWT:10360NEWT:1211601NEWT:876138NEWT:47664NEWT:317548NEWT:3654NEWT:237561NEWT:5833NEWT:6928NEWT:10036NEWT:36745NEWT:498019NEWT:1351NEWT:1438992NEWT:1352NEWT:2649997NEWT:272563NEWT:1147161NEWT:224326NEWT:1333499NCBITaxon:79857NEWT:1096976NEWT:82688NEWT:95648NEWT:3885NEWT:3888NEWT:5821NEWT:1589NEWT:44139NEWT:135622NCBITaxon:4896NEWT:6915NEWT:3649NEWT:101510NEWT:28903NEWT:3880NEWT:272559NEWT:28909NEWT:515849NEWT:3641NEWT:383379NEWT:466585NEWT:10029NEWT:913645NEWT:1000589NEWT:85963NEWT:85962NEWT:143361NEWT:317447NEWT:4688NEWT:7955NEWT:7959NEWT:2261NEWT:31156NEWT:398580NEWT:4442NEWT:4565NEWT:1264690NEWT:515619NEWT:192875NEWT:34305NEWT:59729NEWT:2164133NCBITaxon:183674NEWT:1316931NEWT:224308NEWT:167387NEWT:84645NEWT:44586NEWT:626528NEWT:3347NEWT:511145NEWT:139927NEWT:4558NEWT:209285NEWT:5888NEWT:211586NEWT:747078NEWT:1282NEWT:1283NEWT:431241NEWT:931281NEWT:4550NEWT:1000561NEWT:2133NEWT:294381NEWT:197NEWT:1390363NEWT:77133NEWT:288705NEWT:29176NEWT:145481NCBITaxon:79824NEWT:4787NCBITaxon:4563NEWT:5755NEWT:44689NEWT:3218NEWT:5759NEWT:1736231NEWT:556182NEWT:1270NEWT:374990NEWT:1392NEWT:498217NEWT:156471NEWT:2242NEWT:4784NEWT:11320NEWT:360106NEWT:156476NEWT:286NEWT:391619NEWT:360104NEWT:246196NEWT:287NEWT:246197NEWT:10117NEWT:10239NEWT:10116NEWT:1280NEWT:1735272NEWT:83334NEWT:83332NEWT:44685NEWT:317513NEWT:161934NEWT:1148NEWT:580240NEWT:5508NEWT:294128NEWT:5507NEWT:11676NEWT:55571NEWT:35500NEWT:1140NEWT:100226NEWT:4530NEWT:1143NEWT:4896NEWT:75058NEWT:13616NEWT:1390NEWT:11557NEWT:1094343NEWT:1336795NEWT:172NEWT:644042NEWT:296543NEWT:316435NEWT:42157NEWT:1773NEWT:1895NEWT:1182590NEWT:3712NEWT:82380NEWT:1034304NEWT:105023NEWT:866628NEWT:215358NEWT:935293NEWT:64152NEWT:4924NEWT:749200NEWT:375146NEWT:9378NEWT:990346NEWT:145953NEWT:257309NEWT:100816NEWT:263NEWT:230741NEWT:52283NEWT:284812NEWT:8175NCBITaxon:1313NEWT:43330NEWT:1603293NEWT:408169NEWT:44544NEWT:47946NEWT:4911NEWT:645463NEWT:3702NEWT:129249NEWT:1077286NEWT:943129NEWT:243277NEWT:990119NEWT:7067NEWT:2850NEWT:1111708NEWT:408172NEWT:227321NEWT:408170NEWT:493760NEWT:106590NEWT:260710NEWT:257313NEWT:3827NEWT:400772NEWT:3708NEWT:128161NEWT:332648NEWT:930945NEWT:106592NEWT:536231NEWT:46704NEWT:1436733NEWT:460519NEWT:1187947NEWT:572307NEWT:1432138NEWT:269796NEWT:10312NEWT:1424507NCBITaxon:1773NEWT:1194599NEWT:272844NEWT:54571NEWT:9598NEWT:8030NEWT:9483NEWT:1513458NCBITaxon:40559NEWT:1639NEWT:84588NEWT:188229NEWT:3818NEWT:480NEWT:4909NEWT:67767NEWT:759272NEWT:432359NEWT:46835NEWT:1182263NEWT:109757NEWT:943146NEWT:2711NEWT:300852NEWT:537457NEWT:1502NEWT:376686NEWT:95486NEWT:9103NEWT:508771NEWT:1883446NEWT:29159NEWT:253NEWT:10306NCBITaxon:2759NEWT:1233435NEWT:93061NEWT:8022NEWT:145943NCBITaxon:4932NEWT:595536NEWT:240906NEWT:593117NEWT:89920NEWT:29158NEWT:644223NEWT:3635NEWT:5811NEWT:235443NEWT:180923NEWT:108458NEWT:272623NEWT:272624NEWT:411483NEWT:884019NEWT:198215NEWT:411490NEWT:983964NEWT:118499NEWT:169963NEWT:32644NEWT:527796NEWT:225117NEWT:746128NEWT:499175NEWT:109779NEWT:43665NEWT:1266464NEWT:1715989NCBITaxon:4751NEWT:476272NEWT:3747NEWT:195051NEWT:3988NEWT:367830NEWT:1255228NEWT:178616NEWT:649908NEWT:410289NEWT:373153NEWT:375451NEWT:352472NEWT:357NEWT:1071661NEWT:360094NEWT:470NEWT:41364NEWT:1313NEWT:411469NEWT:84023NEWT:1679NEWT:559292NEWT:39491NCBITaxon:5811NEWT:29491NEWT:411464NEWT:411460NEWT:2014887NEWT:2762NEWT:1174673NEWT:1328426NEWT:562NEWT:411470NEWT:33952NEWT:2094720NCBITaxon:2697049NEWT:571256NCBITaxon:138NEWT:40483NEWT:28038NEWT:1663NEWT:1423NEWT:4932NEWT:3603NEWT:2759NEWT:3847NEWT:38293NEWT:1428NEWT:327159NEWT:178876NEWT:1660NEWT:327160NEWT:573NEWT:9031NEWT:1872122NEWT:7091NEWT:108931NEWT:241368NEWT:1055524NEWT:42528NEWT:190802NEWT:9778NEWT:150475NEWT:303NEWT:9417NEWT:7111NEWT:347515NEWT:1216979NEWT:7237NEWT:9534NEWT:5180NEWT:256737NEWT:9541NEWT:115104NEWT:1121114NEWT:663NEWT:1081927NEWT:1238993NEWT:67825NEWT:187878NEWT:185579NEWT:941442NEWT:220668NEWT:13076NEWT:1821314NEWT:1249668NEWT:7108NEWT:40479NEWT:317NEWT:9646NEWT:5286NEWT:7227NEWT:7469NEWT:885318NEWT:9402NEWT:9644NEWT:415540NEWT:550NEWT:675060NEWT:4081NEWT:334542NEWT:273068NEWT:313625NEWT:554NEWT:27592NEWT:98334NEWT:426428NEWT:451516NEWT:63459NEWT:1276815NEWT:36185NEWT:588858NEWT:1225786NEWT:9639NCBITaxon:11071NEWT:242231NEWT:7574NEWT:1715256NEWT:197221NEWT:7215NEWT:575412NEWT:929793NEWT:29204NEWT:28112NEWT:2172103NEWT:6494NEWT:7460NEWT:6491NEWT:507601NEWT:434271NEWT:747679NEWT:643680NCBITaxon:6157NEWT:42752NEWT:746360NEWT:6239NEWT:89764NEWT:470150NEWT:162425NEWT:216257NEWT:102169NEWT:543734NEWT:9986NEWT:4054NEWT:73239NEWT:38018NEWT:226186NEWT:1268063NEWT:8782NEWT:1263854NEWT:118503NEWT:2059687NEWT:115357NEWT:435590NEWT:1902NEWT:160488NEWT:28104NEWT:1908NEWT:13164NEWT:216129NCBITaxon:2NEWT:985076NEWT:1215323NEWT:2664179NEWT:986NEWT:623NEWT:52641NEWT:7038NEWT:7159NEWT:6192NEWT:28532NCBITaxon:38727NEWT:353152NEWT:2829NEWT:27448NEWT:366581NEWT:216599NEWT:216595NEWT:1194669NEWT:51329NEWT:40559NEWT:243230NEWT:8355NEWT:633NEWT:9685NEWT:7029NEWT:1080772NEWT:8479NEWT:1283300NEWT:186990NEWT:632NEWT:6183NEWT:6063NEWT:630NEWT:49240NEWT:334747NEWT:61235NEWT:343691NEWT:104395NEWT:15368NEWT:6289NEWT:1491466NEWT:436486NEWT:6287NEWT:300641NEWT:727NEWT:9796NEWT:725NEWT:170187NEWT:469008NEWT:260707NEWT:256318NEWT:206411NCBITaxon:6191NEWT:596153NEWT:1836NEWT:185431NEWT:29760NEWT:260704NEWT:1719NEWT:703612NEWT:260705NEWT:80863NEWT:692672NEWT:2697049NEWT:105231NEWT:1216981NEWT:52638NEWT:7004NEWT:4097NCBITaxon:6073NEWT:884204NEWT:9785NEWT:6279NEWT:1123869NEWT:9544NEWT:1329349NEWT:7370NEWT:83906NEWT:607699NCBITaxon:780NCBITaxon:1502NEWT:6282NEWT:208963NEWT:208964NEWT:27337NEWT:1134506NEWT:575584NEWT:38783NEWT:8727NEWT:4006NEWT:8726NEWT:6426NCBITaxon:5476NEWT:6669NEWT:10090NEWT:9935NEWT:89453NEWT:4120NEWT:51515NEWT:5693NEWT:2186NEWT:8724NEWT:51511NEWT:8845NEWT:92867NEWT:8723NEWT:92866NEWT:5334NEWT:37334NEWT:51750NEWT:202950NEWT:382352NEWT:413071NCBITaxon:10359NEWT:1203258NEWT:242619NEWT:160791NEWT:632957NEWT:34073NEWT:373995NEWT:5689NEWT:544404NEWT:9925NEWT:89462NEWT:8839NEWT:4232NEWT:2758385NEWT:37349NEWT:4113NEWT:837NEWT:11298NEWT:171101NEWT:714NEWT:36015NEWT:421932NEWT:312017NEWT:196627NEWT:5691NEWT:48479NEWT:627025NEWT:1097677NEWT:375939NEWT:61674NEWT:870435NEWT:1117957NEWT:5315NEWT:6526NEWT:9913NEWT:8709NEWT:105841NEWT:337330NEWT:4100NEWT:2666255NEWT:1159556NEWT:5671NEWT:2285NEWT:1076NEWT:4101NEWT:6763NEWT:803NEWT:29722NEWT:1392998NEWT:380394NEWT:1692259NEWT:1486917NEWT:419947NEWT:981087NEWT:180066NEWT:226900NEWT:468911NEWT:817NEWT:7604NEWT:818NEWT:135588NEWT:1843183NEWT:5661NEWT:563041NEWT:29719NEWT:58002NEWT:326423NEWT:452467NEWT:36111NEWT:326424NEWT:6997NEWT:452589NEWT:4577NEWT:1416333NEWT:5664NEWT:2157NEWT:1678078NEWT:749906NEWT:418985NEWT:146479NEWT:749907NEWT:1042986NEWT:1911079NEWT:69373NEWT:206189NEWT:145263NEWT:264203NEWT:1480154NEWT:694569NEWT:1274414NEWT:27606NEWT:9739NEWT:59202NEWT:9975NEWT:9612NEWT:38626NEWT:38865NEWT:51953NEWT:3077NEWT:3197NEWT:9615NEWT:10299NEWT:860688NEWT:112273NEWT:36329NEWT:1147787NEWT:62615NCBITaxon:3044782NEWT:1440772NEWT:72407NEWT:1355477NEWT:349741NEWT:9605NEWT:9606NEWT:90371NEWT:157295NEWT:641501NEWT:4153NEWT:2096NEWT:7668NEWT:915099NEWT:74940NEWT:13551NEWT:9721NEWT:137221NEWT:9600NEWT:9963NEWT:1450511NEWT:1143193NEWT:1617910NEWT:174934NEWT:565050NEWT:2823525NEWT:2785785NEWT:411901NEWT:9838NEWT:105884NCBITaxon:9615NEWT:9839NEWT:58334NEWT:1193501NEWT:3055NEWT:58331NEWT:4146NEWT:5478NEWT:6326NEWT:6689NEWT:5476NEWT:1450520NEWT:1042311NEWT:299767NEWT:28835NEWT:1274432NEWT:2053667NEWT:1274426NEWT:70448NEWT:9825NEWT:1274423NEWT:393765NEWT:698936NEWT:39946NEWT:9823NEWT:9940NEWT:95602NEWT:8732NEWT:314539NEWT:521001NEWT:192011NEWT:1813887NEWT:39947NEWT:224911NEWT:718274NEWT:1274420NCBITaxon:11552NEWT:578458