Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces cancer cell death and contributes to tumor rejection by cytotoxic lymphocytes in cancer immunosurveillance and immunotherapy. TRAIL and TRAIL receptor agonists have garnered wide popularity as promising agents for cancer therapy. We previously demonstrated that the loss of fucosylation in cancer cells impairs TRAIL sensitivity; however, the precise structures of the fucosylated glycans that regulate TRAIL sensitivity and their carrier molecules remain elusive. Herein, we observed that Lewis glycans among various fucosylated glycans positively regulate TRAIL-induced cell death. Specifically, Lewis glycans on lacto/neolacto glycosphingolipids, but not glycoproteins including TRAIL receptors, enhanced TRAIL-induced formation of the cytosolic caspase 8 complex, without affecting the formation of the membranous receptor complex. Furthermore, type I Lewis glycan expression in colon cancer cell lines and patient-derived cancer organoids was positively correlated with TRAIL sensitivity. These findings provide novel insights into the regulatory mechanism of TRAIL-induced cell death and facilitate the identification of novel predictive biomarkers for TRAIL-related cancer therapies in future.
This is a preview of subscription content, access via your institution
Subscribe to Journal
Get full journal access for 1 year
only $2.38 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
The authors declare that the data supporting the findings of this study are available within the paper and its supplementary information files. Data not included are available from the corresponding authors upon reasonable request.
Lemoine J, Ruella M, Houot R. Overcoming intrinsic resistance of cancer cells to CAR T-cell killing. Clin Cancer Res. 2021;27:6298–306.
Lemke J, von Karstedt S, Zinngrebe J, Walczak H. Getting TRAIL back on track for cancer therapy. Cell Death Differ. 2014;21:1350–64.
Deng D, Shah K. TRAIL of hope meeting resistance in cancer. Trends Cancer. 2020;6:989–1001.
Dickens LS, Boyd RS, Jukes-Jones R, Hughes MA, Robinson GL, Fairall L, et al. A death effector domain chain DISC model reveals a crucial role for caspase-8 chain assembly in mediating apoptotic cell death. Mol Cell. 2012;47:291–305.
Fox JL, Hughes MA, Meng X, Sarnowska NA, Powley IR, Jukes-Jones R, et al. Cryo-EM structural analysis of FADD:Caspase-8 complexes defines the catalytic dimer architecture for co-ordinated control of cell fate. Nat Commun. 2021;12:819.
Henry CM, Martin SJ. Caspase-8 acts in a non-enzymatic role as a scaffold for assembly of a pro-inflammatory “FADDosome” complex upon TRAIL stimulation. Mol Cell. 2017;65:715–29.
Jin Z, El-Deiry WS. Distinct signaling pathways in TRAIL- versus tumor necrosis factor-induced apoptosis. Mol Cell Biol. 2006;26:8136–48.
Lafont E, Kantari-Mimoun C, Draber P, De Miguel D, Hartwig T, Reichert M, et al. The linear ubiquitin chain assembly complex regulates TRAIL-induced gene activation and cell death. Embo J. 2017;36:1147–66.
Lavrik IN, Mock T, Golks A, Hoffmann JC, Baumann S, Krammer PH. CD95 stimulation results in the formation of a novel death effector domain protein-containing complex. J Biol Chem. 2008;283:26401–8.
Varfolomeev E, Maecker H, Sharp D, Lawrence D, Renz M, Vucic D, et al. Molecular determinants of kinase pathway activation by Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand. J Biol Chem. 2005;280:40599–608.
Lafont E, Hartwig T, Walczak H. Paving TRAIL’s path with ubiquitin. Trends Biochemical Sci. 2018;43:44–60.
Ma B, Simala-Grant JL, Taylor DE. Fucosylation in prokaryotes and eukaryotes. Glycobiology. 2006;16:158R–184R.
Holdener BC, Haltiwanger RS. Protein O-fucosylation: structure and function. Curr Opin Struct Biol. 2019;56:78–86.
Miyoshi E, Moriwaki K, Nakagawa T. Biological function of fucosylation in cancer biology. J Biochem. 2008;143:725–9.
Moriwaki K, Noda K, Furukawa Y, Ohshima K, Uchiyama A, Nakagawa T, et al. Deficiency of GMDS leads to escape from NK cell-mediated tumor surveillance through modulation of TRAIL signaling. Gastroenterology. 2009;137:188–98.
Moriwaki K, Shinzaki S, Miyoshi E. GDP-mannose-4,6-dehydratase (GMDS) deficiency renders colon cancer cells resistant to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor- and CD95-mediated apoptosis by inhibiting complex II formation. J Biol Chem. 2011;286:43123–33.
Wagner KW, Punnoose EA, Januario T, Lawrence DA, Pitti RM, Lancaster K, et al. Death-receptor O-glycosylation controls tumor-cell sensitivity to the proapoptotic ligand Apo2L/TRAIL. Nat Med. 2007;13:1070–7.
Zhang B, van Roosmalen IAM, Reis CR, Setroikromo R, Quax WJ. Death receptor 5 is activated by fucosylation in colon cancer cells. FEBS J. 2019;286:555–71.
Yago K, Zenita K, Ginya H, Sawada M, Ohmori K, Okuma M, et al. Expression of alpha-(1,3)-fucosyltransferases which synthesize sialyl Le(x) and sialyl Le(a), the carbohydrate ligands for E- and P-selectins,in human malignant cell lines. Cancer Res. 1993;53:5559–65.
Costache M, Apoil PA, Cailleau A, Elmgren A, Larson G, Henry S, et al. Evolution of fucosyltransferase genes in vertebrates. J Biol Chem. 1997;272:29721–8.
Pan L, Fu TM, Zhao W, Zhao L, Chen W, Qiu C, et al. Higher-order clustering of the transmembrane anchor of DR5 drives signaling. Cell 2019;176:1477–89.
Moriwaki K, Chan FK. RIP3: a molecular switch for necrosis and inflammation. Genes Dev. 2013;27:1640–9.
Dufour F, Rattier T, Shirley S, Picarda G, Constantinescu AA, Morle A, et al. N-glycosylation of mouse TRAIL-R and human TRAIL-R1 enhances TRAIL-induced death. Cell Death Differ. 2017;24:500–10.
Jiang Y, Wen T, Yan R, Kim SR, Stowell SR, Wang W, et al. O-glycans on death receptors in cells modulate their sensitivity to TRAIL-induced apoptosis through affecting on their stability and oligomerization. FASEB J. 2020;34:11786–801.
Seko A, Yamashita K. Activation of beta1,3-N-acetylglucosaminyltransferase-2 (beta3Gn-T2) by beta3Gn-T8. Possible involvement of beta3Gn-T8 in increasing poly-N-acetyllactosamine chains in differentiated HL-60 cells. J Biol Chem. 2008;283:33094–100.
Togayachi A, Akashima T, Ookubo R, Kudo T, Nishihara S, Iwasaki H, et al. Molecular cloning and characterization of UDP-GlcNAc:lactosylceramide beta 1,3-N-acetylglucosaminyltransferase (beta 3Gn-T5), an essential enzyme for the expression of HNK-1 and Lewis X epitopes on glycolipids. J Biol Chem. 2001;276:22032–40.
Kondo J, Endo H, Okuyama H, Ishikawa O, Iishi H, Tsujii M, et al. Retaining cell-cell contact enables preparation and culture of spheroids composed of pure primary cancer cells from colorectal cancer. Proc Natl Acad Sci USA. 2011;108:6235–40.
Liu C, Deng S, Jin K, Gong Y, Cheng H, Fan Z, et al. Lewis antigennegative pancreatic cancer: An aggressive subgroup. Int J Oncol. 2020;56:900–8.
Nakayama K, Moriwaki K, Imai T, Shinzaki S, Kamada Y, Murata K, et al. Mutation of GDP-mannose-4,6-dehydratase in colorectal cancer metastasis. PLoS One. 2013;8:e70298.
Togayachi A, Kozono Y, Ikehara Y, Ito H, Suzuki N, Tsunoda Y, et al. Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation. Proc Natl Acad Sci USA. 2010;107:11900–5.
Linderoth E, Pilia G, Mahajan NP, Ferby I. Activated Cdc42-associated kinase 1 (Ack1) is required for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor recruitment to lipid rafts and induction of cell death. J Biol Chem. 2013;288:32922–31.
Marconi M, Ascione B, Ciarlo L, Vona R, Garofalo T, Sorice M, et al. Constitutive localization of DR4 in lipid rafts is mandatory for TRAIL-induced apoptosis in B-cell hematologic malignancies. Cell Death Dis. 2013;4:e863.
Ouyang W, Yang C, Zhang S, Liu Y, Yang B, Zhang J, et al. Absence of death receptor translocation into lipid rafts in acquired TRAIL-resistant NSCLC cells. Int J Oncol. 2013;42:699–711.
Akazawa Y, Mott JL, Bronk SF, Werneburg NW, Kahraman A, Guicciardi ME, et al. Death receptor 5 internalization is required for lysosomal permeabilization by TRAIL in malignant liver cell lines. Gastroenterology 2009;136:2365–76.
Kohlhaas SL, Craxton A, Sun XM, Pinkoski MJ, Cohen GM. Receptor-mediated endocytosis is not required for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. J Biol Chem. 2007;282:12831–41.
Song JJ, Szczepanski MJ, Kim SY, Kim JH, An JY, Kwon YT, et al. c-Cbl-mediated degradation of TRAIL receptors is responsible for the development of the early phase of TRAIL resistance. Cell Signal. 2010;22:553–63.
Dufva O, Koski J, Maliniemi P, Ianevski A, Klievink J, Leitner J, et al. Integrated drug profiling and CRISPR screening identify essential pathways for CAR T-cell cytotoxicity. Blood. 2020;135:597–609.
Singh N, Lee YG, Shestova O, Ravikumar P, Hayer KE, Hong SJ, et al. Impaired death receptor signaling in leukemia causes antigen-independent resistance by inducing CAR T-cell dysfunction. Cancer Disco. 2020;10:552–67.
Stern HM, Padilla M, Wagner K, Amler L, Ashkenazi A. Development of immunohistochemistry assays to assess GALNT14 and FUT3/6 in clinical trials of dulanermin and drozitumab. Clin Cancer Res. 2010;16:1587–96.
Moriwaki K, Noda K, Nakagawa T, Asahi M, Yoshihara H, Taniguchi N, et al. A high expression of GDP-fucose transporter in hepatocellular carcinoma is a key factor for increases in fucosylation. Glycobiology 2007;17:1311–20.
Noda K, Miyoshi E, Gu J, Gao CX, Nakahara S, Kitada T, et al. Relationship between elevated FX expression and increased production of GDP-L-fucose, a common donor substrate for fucosylation in human hepatocellular carcinoma and hepatoma cell lines. Cancer Res. 2003;63:6282–9.
We thank Ayaka Hiraishi for technical assistance.
This study was supported by the Japan Society for the Promotion of Science (KM: 16H06945 and 19K07399), the Takeda Science Foundation (KM), GSK Japan Research Grant 2020 (KM), and the Japan Agency for Medical Research and Development (JP20ae0101042 to KM and JP20gm1210002 to HN).
The authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Fukuoka, T., Moriwaki, K., Takamatsu, S. et al. Lewis glycosphingolipids as critical determinants of TRAIL sensitivity in cancer cells. Oncogene 41, 4385–4396 (2022). https://doi.org/10.1038/s41388-022-02434-3