# Globo H ceramide confers chemoresistance and poor prognosis to advanced gallbladder cancer via A2AR/cAMP/PKA pathway **Authors:** Tsai-Hsien Hung, Jung-Tung Hung, Wen-Kuan Huang, Yenlin Huang, Chiao-En Wu, Jing-Yan Cheng, Chien-Wei Lee, John Yu, Yi-Chia Wu, Chun-Nan Yeh, Alice L. Yu PMC · DOI: 10.7150/thno.126092 · 2026-02-26 ## TL;DR Globo H ceramide helps gallbladder cancer resist chemotherapy, but targeting it with specific treatments may improve outcomes. ## Contribution This study identifies Globo H ceramide as a driver of chemoresistance in gallbladder cancer and proposes targeting it to enhance chemotherapy effectiveness. ## Key findings - High Globo H expression correlates with shorter survival in gallbladder cancer patients receiving gemcitabine. - Targeting Globo H with mAb VK9 or vaccines improves chemotherapy efficacy in preclinical models. - Globo H promotes chemoresistance via A2AR/cAMP/PKA-mediated ABCG2 upregulation. ## Abstract Gallbladder cancer (GBC) has poor prognosis, is primarily treated with gemcitabine-based chemotherapy, which is limited by gemcitabine correlates with GBC progression. This study investigate the role of Globo H ceramide (GHCer) in GR and explore whether targeting Glob H could overcome resistance. Globo H expression was assessed by immunohistochemistry in 81 GBC samples. GHCer-induced GR and ABCG2 upregulation were assessed in GBC cell lines, patient-derived xenograft (PDX), and a thioacetamide (TAA)-induced rat cholangiocarcinoma model. A2AR/cAMP/PKA signaling involvement was examined using inhibitors and siRNA. The efficacy of anti-Globo H antibody (mAb VK9) or vaccine (OBI-833/OBI-821) combined with gemcitabine was evaluated in vitro and in vivo. Immune responses were assessed by ELISA and multiplex immunohistochemistry. High Globo H expression correlated with shorter survival in GBC patients receiving gemcitabine. GHCer promoted GR via A2AR/cAMP/PKA-mediated ABCG2 upregulation, which was reversed by mAb VK9 or pathway inhibition. mAb VK9 or OBI-833/821 enhanced gemcitabine efficacy in GBC PDX and TAA cholangiocarcinoma models. OBI-833/821 induced anti-Globo H IgG/IgM, reduced Foxp3⁺ Tregs, and increased CD161⁺ NK cells in TAA model. A compassionate clinical use of 833/821 led to stable disease. GHCer promotes GR by upregulating ABCG2 via A2AR/cAMP/PKA signaling. Targeting Globo H may improve chemotherapy response in GBC. ## Linked entities - **Proteins:** ADORA2A (adenosine A2a receptor), ABCG2 (ATP binding cassette subfamily G member 2 (JR blood group)), FOXP3 (forkhead box P3), KLRB1 (killer cell lectin like receptor B1) - **Chemicals:** gemcitabine (PubChem CID 60750), cAMP (PubChem CID 6076), thioacetamide (PubChem CID 2723949) - **Diseases:** gallbladder cancer (MONDO:0003220), cholangiocarcinoma (MONDO:0019087) - **Species:** Mus musculus (taxon 10090) ## Full-text entities - **Genes:** ABCB6 (ATP binding cassette subfamily B member 6 (LAN blood group)) [NCBI Gene 10058] {aka ABC, LAN, MTABC3, PRP, umat}, Mtr (5-methyltetrahydrofolate-homocysteine methyltransferase) [NCBI Gene 81522] {aka methioninesynthase}, ABCC6 (ATP binding cassette subfamily C member 6) [NCBI Gene 368] {aka ABC34, ARA, EST349056, GACI2, MLP1, MOAT-E}, ADORA2A (adenosine A2a receptor) [NCBI Gene 135] {aka A2aR, ADORA2, RDC8}, Foxp3 (forkhead box P3) [NCBI Gene 317382] {aka RGD1562112}, ABCC10 (ATP binding cassette subfamily C member 10) [NCBI Gene 89845] {aka EST182763, MRP7, SIMRP7}, Ggh (gamma-glutamyl hydrolase) [NCBI Gene 25455], RRM1 (ribonucleotide reductase catalytic subunit M1) [NCBI Gene 6240] {aka PEOB6, R1, RIR1, RR1}, PGR (progesterone receptor) [NCBI Gene 5241] {aka NR3C3, PR}, Mgat1 (alpha-1,3-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferase) [NCBI Gene 81519], ABCC2 (ATP binding cassette subfamily C member 2) [NCBI Gene 1244] {aka ABC30, CMOAT, DJS, MRP2, cMRP}, RAD51 (RAD51 recombinase) [NCBI Gene 5888] {aka BRCC5, FANCR, HRAD51, HsRad51, HsT16930, MRMV2}, CREB1 (cAMP responsive element binding protein 1) [NCBI Gene 1385] {aka CREB, CREB-1}, DCK (deoxycytidine kinase) [NCBI Gene 1633], TWIST1 (twist family bHLH transcription factor 1) [NCBI Gene 7291] {aka ACS3, BPES2, BPES3, CRS, CRS1, CSO}, ABCC3 (ATP binding cassette subfamily C member 3) [NCBI Gene 8714] {aka ABC31, EST90757, MLP2, MOAT-D, MRP3, cMOAT2}, SLC29A1 (solute carrier family 29 member 1 (Augustine blood group)) [NCBI Gene 2030] {aka AUG, ENT1, hENT1}, AFP (alpha fetoprotein) [NCBI Gene 174] {aka AFPD, FETA, HPAFP}, ABCB7 (ATP binding cassette subfamily B member 7) [NCBI Gene 22] {aka ABC7, ASAT, Atm1p, EST140535}, BCL2L1 (BCL2 like 1) [NCBI Gene 598] {aka BCL-XL/S, BCL2L, BCLX, Bcl-X, PPP1R52}, BCL2 (BCL2 apoptosis regulator) [NCBI Gene 596] {aka Bcl-2, PPP1R50}, CTSA (cathepsin A) [NCBI Gene 5476] {aka BSVD6, GLB2, GSL, NGBE, PPCA, PPGB}, BRAF (B-Raf proto-oncogene, serine/threonine kinase) [NCBI Gene 673] {aka B-RAF1, B-raf, BRAF-1, BRAF1, NS7, RAFB1}, TSNAX (translin associated factor X) [NCBI Gene 7257] {aka C3PO, TRAX}, ABCB5 (ATP binding cassette subfamily B member 5) [NCBI Gene 340273] {aka ABCB5alpha, ABCB5beta, EST422562}, ABCB1 (ATP binding cassette subfamily B member 1) [NCBI Gene 5243] {aka ABC20, CD243, CLCS, ENPAT, GP170, MDR1}, IGHG3 (immunoglobulin heavy constant gamma 3 (G3m marker)) [NCBI Gene 3502] {aka IgG3}, Gem (GTP binding protein overexpressed in skeletal muscle) [NCBI Gene 297902], BRCA1 (BRCA1 DNA repair associated) [NCBI Gene 672] {aka BRCAI, BRCC1, BROVCA1, FANCS, IRIS, PNCA4}, Mgat2 (alpha-1,6-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferase) [NCBI Gene 94273] {aka Gnt2}, MIB1 (MIB E3 ubiquitin protein ligase 1) [NCBI Gene 57534] {aka DIP-1, DIP1, LVNC7, MIB, ZZANK2, ZZZ6}, RRM2 (ribonucleotide reductase regulatory subunit M2) [NCBI Gene 6241] {aka C2orf48, R2, RR2, RR2M}, Abcg2 (ATP binding cassette subfamily G member 2) [NCBI Gene 312382] {aka BCRP1, Bcrp}, ABCB11 (ATP binding cassette subfamily B member 11) [NCBI Gene 8647] {aka ABC16, BRIC2, BSEP, PFIC-2, PFIC2, PGY4}, KLRB1 (killer cell lectin like receptor B1) [NCBI Gene 3820] {aka CD161, CLEC5B, NKR, NKR-P1, NKR-P1A, NKRP1A}, ABCC5 (ATP binding cassette subfamily C member 5) [NCBI Gene 10057] {aka ABC33, EST277145, MOAT-C, MOATC, MRP5, SMRP}, PRKACA (protein kinase cAMP-activated catalytic subunit alpha) [NCBI Gene 5566] {aka CAFD1, PKACA, PPNAD4}, ABCC1 (ATP binding cassette subfamily C member 1 (ABCC1 blood group)) [NCBI Gene 4363] {aka ABC29, ABCC, DFNA77, GS-X, MRP, MRP1}, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 2597] {aka G3PD, GAPD, HEL-S-162eP}, PSG2 (pregnancy specific beta-1-glycoprotein 2) [NCBI Gene 5670] {aka CEA, PSBG2, PSG1}, FOXP3 (forkhead box P3) [NCBI Gene 50943] {aka AIID, DIETER, IPEX, JM2, PIDX, XPID}, ERBB2 (erb-b2 receptor tyrosine kinase 2) [NCBI Gene 2064] {aka CD340, HER-2, HER-2/neu, HER2, MLN 19, MLN-19}, ABCG2 (ATP binding cassette subfamily G member 2 (JR blood group)) [NCBI Gene 9429] {aka ABC15, ABCP, BCRP, BMDP, CD338, CDw338}, MUC4 (mucin 4, cell surface associated) [NCBI Gene 4585] {aka ASGP, HSA276359, MUC-4}, CAMP (cathelicidin antimicrobial peptide) [NCBI Gene 820] {aka CAP-18, CAP18, CRAMP, FALL-39, FALL39, HSD26}, AZIN2 (antizyme inhibitor 2) [NCBI Gene 113451] {aka ADC, AZIB1, ODC-p, ODC1L, ODCp}, TAP1 (transporter 1, ATP binding cassette subfamily B member) [NCBI Gene 6890] {aka ABC17, ABCB2, APT1, D6S114E, MHC1D1, PSF-1}, ABCC4 (ATP binding cassette subfamily C member 4 (PEL blood group)) [NCBI Gene 10257] {aka MOAT-B, MOATB, MRP4}, PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha) [NCBI Gene 5290] {aka CCM4, CLAPO, CLOVE, CWS5, HMH, MCAP}, SLC28A3 (solute carrier family 28 member 3) [NCBI Gene 64078] {aka CNT3}, BRCA2 (BRCA2 DNA repair associated) [NCBI Gene 675] {aka BRCC2, BROVCA2, FACD, FAD, FAD1, FANCD}, ABCC11 (ATP binding cassette subfamily C member 11) [NCBI Gene 85320] {aka EWWD, MRP8, WW}, ABCB4 (ATP binding cassette subfamily B member 4) [NCBI Gene 5244] {aka ABC21, GBD1, ICP3, MDR2, MDR2/3, MDR3} - **Diseases:** mucinous adenocarcinoma (MESH:D002288), gallbladder adenocarcinoma (MESH:D000230), SD (MESH:D060050), cancer (MESH:D009369), BTCs (MESH:D001661), cytotoxicity (MESH:D064420), PDX (MESH:C536408), carcinomatosis (MESH:D002277), metastasis (MESH:D009362), tumorigenic (MESH:D002471), death (MESH:D003643), ADCC (MESH:D007153), bile duct cancer (MESH:D001650), neuroblastoma (MESH:D009447), GBC tumor (MESH:D005706), PR (MESH:D004828), pelvic lesion (MESH:D034161), breast cancer (MESH:D001943), triple-negative breast cancer (MESH:D064726), PD (MESH:D018450), CCA tumor (MESH:D018281) - **Chemicals:** glycan (MESH:D011134), bilirubin (MESH:D001663), Lipofectamine (MESH:C086724), urea (MESH:D014508), glycosphingolipid (MESH:D006028), nitrogen (MESH:D009584), sialic acid (MESH:D019158), FDG (MESH:D019788), dinutuximab (MESH:C112746), streptomycin (MESH:D013307), polyacrylamide (MESH:C016679), Dewax (-), cisplatin (MESH:D002945), hydrogen peroxide (MESH:D006861), DHB (MESH:C003870), lactosylceramides (MESH:D007790), paraffin (MESH:D010232), methanol (MESH:D000432), TAA (MESH:D013853), FOLFOX (MESH:C410216), Europium (MESH:D005063), 2,5-dihydroxybenzoic acid (MESH:C010925), penicillin (MESH:D010406), ZM241385 (MESH:C097270), glycolipid (MESH:D006017), N-acetylgalactosamine (MESH:D000116), MES (MESH:C004550), Gemcitabine (MESH:D000093542), MMAE (MESH:C495575), Tween20 (MESH:D011136), PVDF (MESH:C024865), MnCl2 (MESH:C025340), disialoganglioside (MESH:C025447), SDS (MESH:D012967), Formalin (MESH:D005557), DAPI (MESH:C007293), DMSO (MESH:D004121), Cyclic AMP (MESH:D000242), ceramide (MESH:D002518), galactose (MESH:D005690), SP (MESH:C000604007), saponin (MESH:D012503), water (MESH:D014867), GlcCer (MESH:D005963), CO2 (MESH:D002245), chloroform (MESH:D002725), Rh 123 (MESH:D020112), Sepharose (MESH:D012685), H89 (MESH:C063509) - **Species:** Rattus norvegicus (brown rat, species) [taxon 10116], Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606], Oryctolagus cuniculus (domestic rabbit, species) [taxon 9986], Mycoplasma (genus) [taxon 2093] - **Mutations:** P0720L, A2A, P0745L - **Cell lines:** GBC01 — Epinephelus coioides (Orange-spotted grouper), Spontaneously immortalized cell line (CVCL_R900), GBC — Homo sapiens (Human), Gallbladder carcinoma, Cancer cell line (CVCL_IU75), TGCB24 — Mus musculus (Mouse), Hybridoma (CVCL_C5HY), TGBC24 — Homo sapiens (Human), Gallbladder undifferentiated carcinoma, Cancer cell line (CVCL_1769), SNU308 — Homo sapiens (Human), Gallbladder carcinoma, Cancer cell line (CVCL_5048) ## Figures 6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12964375/full.md --- Source: https://tomesphere.com/paper/PMC12964375