doi: https://doi.org/10.7150/thno.123973
https://www.thno.org/ms/acceptms
Abstract
Rationale: Glycosylation of drug delivery vehicles enables selective tumor microenvironment (TME) targeting but is limited by the lack of precise glycan control and unbiased evaluation of in situ targeting. We developed a clickable albumin nanoplatform engineered by distinct glycosylation for selective in vivo cell targeting (CAN-DGIT) with a defined number of sugar moieties and integrated spatial transcriptomics (ST) to map nanoparticle–TME interactions.
Methods: Albumin was functionalized with azadibenzocyclooctyne (ADIBO) at a controlled degree of functionalization (DOF), confirmed by MALDI-TOF and UV–vis spectroscopy, followed by conjugation of azide-functionalized mannose, galactose, or glucose via click chemistry. Nanoparticles were labeled with 64Cu or fluorescent dyes for PET imaging and ex vivo analysis in healthy and 4T1 tumor-bearing mice. ST based algorithms, spatial gene–image integration (SPADE), cell-type deconvolution (CellDART), and image-based molecular signature analysis (IAMSAM), were used to define TME clusters, associated cell populations, and glycan receptor gene signatures. Clodronate-loaded glycosylated albumins were tested for tumor-associated macrophage (TAM)depletion.
Results: Glycosylation type of CAN-DGIT dictated pharmacokinetics and targeting. Mannosylated albumin (Man-Alb) showed rapid hepatic retention via mannose receptors on Kupffer cells and TAMs; galactosylated albumin (Gal-Alb) exhibited rapid hepatobiliary clearance with the highest tumor-to-liver ratio; glucosylated albumin at the C6 position (Glc(6)-Alb) progressively accumulated in tumors, correlating with glucose transporter 1(GLUT1)-expressing cancer cells. ST confirmed Man-Alb enrichment in extracellular matrix (ECM)/TAM-rich clusters (mannose receptor C-type 1, Mrc1-high) and Gal-/Glc-Alb uptake in glycolytic/hypoxic tumor clusters (Slc2a1-high). Man-Alb–clodronate achieved potent CD206^+ TAM depletion without altering drug release kinetics.
Conclusions: Precisely tuned glycosylation enables programmable biodistribution and cell-type targeting of albumin nanoparticles in the TME. Integrating PET with ST provides a robust framework for mechanistic mapping of nanomedicine uptake. The CAN-DGIT platform offers a versatile strategy for developing targeted theranostic agents with immunomodulatory potential.
Authors
Park JY, Choi J, Park J, Kim JS, Ann YC, Chung H, Park J, Kim J, Seok SH, Choi H, Im H-J, Lee Y-S
