Rituximab-Conjugated DART Nanoformulation for CD20-Targeted Drug Delivery in Non-Hodgkin Lymphoma.
To develop rituximab (RTX)-conjugated decreased nonspecific adhesivity receptor-targeted (DART) nanoparticles for CD20-targeted paclitaxel (PTX) delivery in non-Hodgkin lymphoma (NHL) and evaluate their targeting, therapeutic activity, formulation stability, and immune-associated cellular responses.
PTX-loaded PLGA-PEG nanoparticles were conjugated with RTX (RTX-DART/PTX) or control IgG (IgG-DART/PTX). Physicochemical properties and colloidal and frozen-storage stability were characterized. CD20-dependent cellular association and intracellular localization were evaluated in Raji lymphoma cells using flow cytometry, competitive blocking, and confocal microscopy. Cytotoxicity was assessed following short-term treatment and media replacement. Calreticulin (CRT) surface exposure and macrophage polarization-associated markers were evaluated in vitro. Anti-tumor efficacy was assessed in a systemic luciferase-expressing Raji xenograft model.
RTX-DART/PTX exhibited a size near 100 nm, low polydispersity, near-neutral surface charge, and 7-8% PTX loading. DART nanoparticles maintained their colloidal properties in serum incubation for up to 72 hours, while frozen storage at -20°C in 10% sucrose for 3 weeks preserved physicochemical properties and biological activity. RTX-DART showed greater CD20-dependent cellular association and intracellular localization than IgG-DART and free RTX blocking reduced nanoparticle association. Under short-exposure conditions, RTX-DART/PTX produced greater cytotoxicity than free PTX and IgG-DART/PTX. RTX-DART/PTX also enhanced CRT surface exposure, while PTX-containing treatments altered macrophage polarization-associated markers and IGF1 secretion in vitro. In vivo, RTX-DART/PTX reduced systemic tumor bioluminescence and improved survival relative to PBS and IgG-DART/PTX.
These findings support RTX-DART/PTX as a proof-of-concept CD20-targeted nanomedicine strategy for systemic NHL. Further validation in additional lymphoma models, immunocompetent or humanized systems, and pharmacokinetic and biodistribution studies is required.
PTX-loaded PLGA-PEG nanoparticles were conjugated with RTX (RTX-DART/PTX) or control IgG (IgG-DART/PTX). Physicochemical properties and colloidal and frozen-storage stability were characterized. CD20-dependent cellular association and intracellular localization were evaluated in Raji lymphoma cells using flow cytometry, competitive blocking, and confocal microscopy. Cytotoxicity was assessed following short-term treatment and media replacement. Calreticulin (CRT) surface exposure and macrophage polarization-associated markers were evaluated in vitro. Anti-tumor efficacy was assessed in a systemic luciferase-expressing Raji xenograft model.
RTX-DART/PTX exhibited a size near 100 nm, low polydispersity, near-neutral surface charge, and 7-8% PTX loading. DART nanoparticles maintained their colloidal properties in serum incubation for up to 72 hours, while frozen storage at -20°C in 10% sucrose for 3 weeks preserved physicochemical properties and biological activity. RTX-DART showed greater CD20-dependent cellular association and intracellular localization than IgG-DART and free RTX blocking reduced nanoparticle association. Under short-exposure conditions, RTX-DART/PTX produced greater cytotoxicity than free PTX and IgG-DART/PTX. RTX-DART/PTX also enhanced CRT surface exposure, while PTX-containing treatments altered macrophage polarization-associated markers and IGF1 secretion in vitro. In vivo, RTX-DART/PTX reduced systemic tumor bioluminescence and improved survival relative to PBS and IgG-DART/PTX.
These findings support RTX-DART/PTX as a proof-of-concept CD20-targeted nanomedicine strategy for systemic NHL. Further validation in additional lymphoma models, immunocompetent or humanized systems, and pharmacokinetic and biodistribution studies is required.
Authors
Mahmud Mahmud, Kong Kong, Lee Lee, Pinzon Burgos Pinzon Burgos, Heredia Heredia, Suk Suk, Kim Kim
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