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PLGA Nanoparticles Encapsulating Nucleolin Peptide: A Novel
Peptide Vaccine Strategy for Triple Negative Breast Cancer
Sirawit Jirawannaporn1, Suyanee Thongchot1,2, Kamonlatth Rodponthukwaji3,4, Peti Thuwajit1
,
Primana Punnakitikashem3,4, Chanitra Thuwajit1*
1 Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University
2 Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Research Department,
Faculty of Medicine, Siriraj Hospital, Mahidol University
3 Department of Biochemistry, Faculty of Medicine, Siriraj Hospital, Mahidol University
4 Siriraj Center of Research Excellence in Theranostic Nanomedicine, Faculty of Medicine, Siriraj Hospital,
Mahidol University
*Corresponding Author E-mail: cthuwajit@yahoo.com
Background: Abstract
Triple Negative Breast Cancer (TNBC) is an aggressive subtype with limited therapeutic
options and poor prognosis. Dendritic cell (DC)-based immunotherapy is a promising
approach for TNBC, but its success depends on the efficient delivery of tumor-specific
antigens. This study aims to develop poly(lactic-co-glycolic acid) (PLGA)-based
nanoparticles (PLGA-NPs) encapsulating NCL-01, a tumor-associated peptide from
Nucleolin that overexpressed in TNBC, to stimulate DCs and elicit specific cytotoxic T
lymphocytes against cancer cells.
Methods: Results: Conclusion: PLGA-NPs encapsulated NCL-01 peptide (pNCL-01-NPs) with sequence KMAPPPKEV
were synthesized using a double emulsion solvent evaporation . The nanoparticles were
characterized for size, surface charge, encapsulation efficiency, and peptide release
kinetics. DCs were pulsed with pNCL-01-NPs and co-cultured with autologous T cells.
Phenotypic maturation of DCs and T cell activation were analyzed by flow cytometry.
Antitumor activity was evaluated using 2D crystal violet cytotoxicity and 3D spheroid
killing assays against NCL-01-positive MDA-MB-231 TNBC cells and normal MCF-10A
breast epithelial cells.
The pNCL-01-NPs had a mean diameter of 137.73 ± 27.69 nm, a zeta potential of −26.68 ±
0.87 mV, and encapsulation efficiency of 8.97 ± 1.89%. Sustained peptide release reached
47.10 ± 11.39% at 48 hours. DCs pulsed with pNCL-01-NPs showed a significant increase in
the overall upregulation of maturation markers, as indicated by the stacked bar graph, with
notable increases in the combined expression of CD40, CD80, CD83, CD86, and HLA-DR
compared to the bare pNCL01 peptide. T cells activated via these DCs produced
higher levels of IFN-γ. In functional assays, the T cells specifically lysed NCL-01-
positive TNBC cells but spared normal cells. 3D assays confirmed significant tumor
spheroid shrinkage and decreased fluorescence intensity.
This study demonstrates that pNCL-01-NPs effectively induce DC maturation and generate
potent tumor-specific T cell responses when compared with bare NCL-01 peptide. The
platform exhibits strong antitumor activity in both 2D and 3D models, supporting its
potential as a safe and effective peptide-based nanovaccine for TNBC immunotherapy.
Harmony in health: Innovation for Sustainable Medicine
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