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Updated: Jun 20, 2026
Mimicking and Manipulating Pancreatic Acinar-to-Ductal Metaplasia in 3-dimensional Cell Culture
Published on: February 11, 2019
Aberrant Glycosylation in Pancreatic Ductal Adenocarcinoma 3D Organoids Is Mediated by KRAS Mutations
Hiromitsu Nakahashi1, Tatsuya Oda1, Osamu Shimomura1
1Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
Abstract:
Aberrant glycosylation in tumor cells is a hallmark during carcinogenesis. KRAS gene mutations are the most well-known oncogenic abnormalities but their association with glycan alterations in pancreatic ductal adenocarcinoma (PDAC) is largely unknown. We employed patient-derived 3D organoids to culture pure live PDAC cells, excluding contamination by fibroblasts and immune cells, to gasp the comprehensive cancer cell surface glycan expression profile using lectin microarray and transcriptomic analyses. Surgical specimens from 24 PDAC patients were digested and embedded into a 3D culture system. Surface-bound glycans of 3D organoids were analyzed by high-density, 96-lectin microarrays. KRAS mutation status and expression of various glycosyltransferases were analyzed by RNA-seq. We successfully established 16 3D organoids: 14 PDAC, 1 intraductal papillary mucinous neoplasm (IPMN), and 1 normal pancreatic duct. KRAS was mutated in 13 (7 G12V, 5 G12D, 1 Q61L) and wild in 3 organoids (1 normal duct, 1 IPMN, 1 PDAC). Lectin reactivity of AAL (Aleuria aurantia) and AOL (Aspergillus oryzae) with binding activity to α1-3 fucose was higher in organoids with KRAS mutants than those with KRAS wild-type. FUT6 (α1-3fucosyltransferase 6) and FUT3 (α1-3/4 fucosyltransferase 3) expression was also higher in KRAS mutants than wild-type. Meanwhile, mannose-binding lectin (rRSL [Ralstonia solanacearum] and rBC2LA [Burkholderia cenocepacia]) signals were higher while those of galactose-binding lectins (rGal3C and rCGL2) were lower in the KRAS mutants. We demonstrated here that PDAC 3D-cultured organoids with KRAS mutations were dominantly covered in increased fucosylated glycans, pointing towards novel treatment targets and/or tumor markers.
Insights
KRAS mutations in pancreatic cancer are linked to altered cell surface glycans. This study found increased fucosylated glycans on KRAS-mutant pancreatic ductal adenocarcinoma cells, suggesting new therapeutic targets.
Area of Science:
- Oncology
- Glycobiology
- Molecular Biology
Background:
- Aberrant glycosylation is a key feature of cancer cells.
- KRAS gene mutations are common in pancreatic ductal adenocarcinoma (PDAC), but their link to glycan changes is unclear.
Purpose of the Study:
- To investigate the relationship between KRAS mutations and cell surface glycan profiles in PDAC.
- To analyze glycan expression in pure cancer cells using patient-derived 3D organoids.
Main Methods:
- Established 16 3D organoids from patient samples (14 PDAC, 1 IPMN, 1 normal).
- Analyzed KRAS mutation status and glycosyltransferase expression via RNA-seq.
- Assessed cell surface glycan profiles using high-density lectin microarrays.
Main Results:
- KRAS mutations were identified in 13 of 16 organoids.
- Organoids with KRAS mutations showed increased reactivity with lectins for alpha-1,3 fucose (AAL, AOL).
- Higher expression of FUT6 and FUT3 was observed in KRAS-mutant organoids, correlating with increased fucosylation and altered mannose/galactose binding lectin signals.
Conclusions:
- KRAS-mutant PDAC cells exhibit a distinct glycan profile characterized by increased fucosylation.
- These findings highlight potential novel therapeutic targets and biomarkers for PDAC.
