Lectin-Assisted Imaging Mass Cytometry (Lectin-IMC) Enables Spatial Validation of Disease-Associated Glyco-Niches within a Multimodal Glycoprotein Analysis Framework
- 1National Institute of Advanced Industrial Science and Technology (AIST)
Abstract
Understanding the pathological significance of protein glycosylation and translating disease-associated glycan alterations into diagnostic and therapeutic opportunities require integrated analysis of glycans, their carrier glycoproteins, and spatial context. Here, we present an expanded multimodal glycoprotein analysis framework incorporating lectin-assisted imaging mass cytometry (Lectin-IMC) for stepwise discovery and spatial validation of disease-associated glyco-niches. Disease-associated glycans identified by laser microdissection-assisted lectin microarray (LMD-LMA) tissue glycome mapping are spatially evaluated by Lectin-IMC in relation to cell types and tissue microenvironments, and candidate carrier glycoproteins identified by MS-based glycoproteomics are subsequently incorporated for higher-order evaluation of glycan-protein-cell type relationships. Multiplex panel design is guided by LMA-based assessment of lectin-lectin interactions and biologically informed probe selection. Using a dilated cardiomyopathy model, we constructed a five-lectin panel centered on Wisteria floribunda agglutinin (WFA), which recognizes fibrosis-associated asialo N-glycans identified previously. Glycan-dependent WFA detection was validated by competitive inhibition and PNGase F treatment, and WFA-reactive glycans were spatially associated with fibrotic regions containing ACTA2VIM myofibroblast-like cells. Among six extracellular matrix glycoprotein candidates identified in WFA-binding fractions, periostin showed the strongest spatial correspondence with WFA-positive regions by pixel-based quantitative analysis and was further associated with ACTA2VIMWFA fibrotic microenvironments. The optimized Lectin-IMC panel was also transferable to a human FFPE cardiomyopathy specimen. Collectively, Lectin-IMC provides intuitive spatial visualization and interpretability of glycan-protein-cell type relationships and serves as an on-tissue spatial validation and prioritization layer within an iterative multimodal framework in which spatially defined glyco-niches can guide subsequent proteomic or glycoproteomic discovery.
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BibTeX
@misc{nagaiokatani2026lectinassisted,
title = {{Lectin-Assisted Imaging Mass Cytometry (Lectin-IMC) Enables Spatial Validation of Disease-Associated Glyco-Niches within a Multimodal Glycoprotein Analysis Framework}},
author = {C. Nagai-Okatani and U. Horiuchi and P. Boottanun and K. Miyako and A. Kuno},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.19.752828},
url = {https://www.biorxiv.org/content/10.64898/2026.09.19.752828v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Lectin-Assisted Imaging Mass Cytometry (Lectin-IMC) Enables Spatial Validation of Disease-Associated Glyco-Niches within a Multimodal Glycoprotein Analysis Framework
AU - C. Nagai-Okatani
AU - U. Horiuchi
AU - P. Boottanun
AU - K. Miyako
AU - A. Kuno
PY - 2026
DA - 2026/09/19
PB - bioRxiv
DO - 10.64898/2026.09.19.752828
UR - https://www.biorxiv.org/content/10.64898/2026.09.19.752828v1
AB - Understanding the pathological significance of protein glycosylation and translating disease-associated glycan alterations into diagnostic and therapeutic opportunities require integrated analysis of glycans, their carrier glycoproteins, and spatial context. Here, we present an expanded multimodal glycoprotein analysis framework incorporating lectin-assisted imaging mass cytometry (Lectin-IMC) for stepwise discovery and spatial validation of disease-associated glyco-niches. Disease-associated glycans identified by laser microdissection-assisted lectin microarray (LMD-LMA) tissue glycome mapping are spatially evaluated by Lectin-IMC in relation to cell types and tissue microenvironments, and candidate carrier glycoproteins identified by MS-based glycoproteomics are subsequently incorporated for higher-order evaluation of glycan-protein-cell type relationships. Multiplex panel design is guided by LMA-based assessment of lectin-lectin interactions and biologically informed probe selection. Using a dilated cardiomyopathy model, we constructed a five-lectin panel centered on Wisteria floribunda agglutinin (WFA), which recognizes fibrosis-associated asialo N-glycans identified previously. Glycan-dependent WFA detection was validated by competitive inhibition and PNGase F treatment, and WFA-reactive glycans were spatially associated with fibrotic regions containing ACTA2VIM myofibroblast-like cells. Among six extracellular matrix glycoprotein candidates identified in WFA-binding fractions, periostin showed the strongest spatial correspondence with WFA-positive regions by pixel-based quantitative analysis and was further associated with ACTA2VIMWFA fibrotic microenvironments. The optimized Lectin-IMC panel was also transferable to a human FFPE cardiomyopathy specimen. Collectively, Lectin-IMC provides intuitive spatial visualization and interpretability of glycan-protein-cell type relationships and serves as an on-tissue spatial validation and prioritization layer within an iterative multimodal framework in which spatially defined glyco-niches can guide subsequent proteomic or glycoproteomic discovery.
ER -