Frontiers in Bioengineering and Biotechnology2020ResearchDrug Delivery

Melanoma Peptide MHC Specific TCR Expressing T-Cell Membrane Camouflaged PLGA Nanoparticles for Treatment of Melanoma Skin Cancer

Serkan Yaman, Harish Ramachandramoorthy, Gizem Oter, Daria Zhukova, Tam Nguyen, Manoj K. Sabnani, Jon A. Weidanz, Kytai T. Nguyen29 September 2026DOI 10.3389/fbioe.2020.00943

Summary

Melanoma is an aggressive skin cancer with limited treatment efficacy due to non-specific drug targeting, severe side effects, and multidrug resistance. Current cell-based immunotherapies are costly, complex, and carry long-term autoimmune risks. There is a need for a targeted, biocompatible drug delivery system that selectively recognizes melanoma cells and provides sustained drug release. Physicochemical properties: T-MNPs (1:2) were 193 ± 56 nm, PDI 0.265, zeta −36 mV; stable in saline for 48 h. Drug loading 61%; sustained trametinib release over 28 days, slowest at highest membrane ratio (1:2). - TCR.

Keywords

T cellsNanoparticlesPLGATCR signalingBiodistributionCellular uptakePolymeric

Purpose: Melanoma is an aggressive skin cancer with limited treatment efficacy due to non-specific drug targeting, severe side effects, and multidrug resistance. Current cell-based immunotherapies are costly, complex, and carry long-term autoimmune risks. There is a need for a targeted, biocompatible drug delivery system that selectively recognizes melanoma cells and provides sustained drug release.

Hypothesis: If PLGA nanoparticles loaded with trametinib are coated with membranes from a T-cell hybridoma (19LF6) expressing an anti-gp100/HLA-A2 T-cell receptor (TCR), then the resulting T-cell membrane-coated nanoparticles (T-MNPs) will specifically bind gp100-presenting melanoma cells, enhance cellular uptake, improve therapeutic efficacy, and increase tumor retention in vivo compared with uncoated or non-specific membrane-coated nanoparticles.

Aims: Primary aim: Develop T-cell membrane-camouflaged PLGA nanoparticles loaded with trametinib (T-MNPs) for targeted melanoma therapy.

Secondary aim 1: Characterize T-MNPs for size, zeta potential, morphology, stability, TCR presence, drug loading, and release kinetics.

Secondary aim 2: Evaluate binding kinetics, cellular uptake, and in vitro cytotoxicity against gp100-positive melanoma cell lines (DM-6, 1520) versus gp100-negative A549 cells.

Secondary aim 3: Assess cyto- and hemo-compatibility of T-MNPs.

Secondary aim 4: Evaluate in vivo biodistribution and tumor retention of T-MNPs in a subcutaneous DM-6 melanoma xenograft mouse model.

Delivery system:

Component: Polymer; Description: PLGA (50:50, carboxyl end groups)

Component: Nanoparticle Type; Description: Single-emulsion (O/W) PLGA nanoparticles

Component: Targeting Ligand; Description: Cell membrane from 19LF6 T-cell hybridoma expressing anti-gp100/HLA-A2 TCR

Component: Payload; Description: Trametinib (FDA-approved MEK inhibitor for BRAF V600E/K melanoma); coumarin-6 or DiD for imaging

Component: Membrane Coating Ratios; Description: NP: membrane protein (w/w) = 1:0.5, 1:1, 1:2, 1:3

Component: Size; Description: T-MNPs (1:2): 193 ± 56 nm; NNPs: 171.7 ± 76 nm

Component: Zeta Potential; Description: T-MNPs: −36 mV; NNPs: −20 mV

Component: Drug Loading Efficiency; Description: 61%

Component: Release; Description: Initial burst followed by sustained release up to 28 days; slower release with higher membrane content

Component: Controls; Description: Naked PLGA NPs (NNPs), DO11.10 membrane-coated NPs (D-MNPs, non-specific T-cell), A549 membrane-coated NPs (A-MNPs, non-specific cancer cell)

Approach: In Vitro: - Cell lines: DM-6 and 1520 (gp100-positive melanoma), A549 (gp100-negative lung cancer), HDF (human dermal fibroblasts), 19LF6 and DO11.10 hybridomas. - Uptake and therapeutic studies: Coumarin-6–loaded NPs at NP:membrane ratios 1:0.5, 1:1, 1:2; concentrations 100–1000 µg/mL for uptake; IC25–IC75 trametinib concentrations for MTS cytotoxicity. - Cyto/hemo compatibility: HDF cells at 50–1000 µg/mL; human blood for clotting and hemolysis assays.

In Vivo: - Model: Subcutaneous DM-6 melanoma xenograft in athymic nude mice; treatment started at ~150 mm³. - Groups (n = 6/group): Saline, PLGA NP, DO11.10 T-MNP (D-MNP), 19LF6 T-MNP (T-MNP). - Administration: Intravenous tail vein injection of DiD-labeled NPs. - Imaging: In vivo at 2, 4, 6 h; ex vivo organs at 24 h.

Key methods:

Technique: Dynamic light scattering (DLS); Purpose: Particle size, PDI, zeta potential

Technique: Transmission electron microscopy (TEM); Purpose: Morphology and core-shell structure

Technique: Flow cytometry; Purpose: Confirm TCR β chain on 19LF6 cells and T-MNPs

Technique: ResoSens label-free detection; Purpose: Binding kinetics to gp100-biotinylated MHC

Technique: Western blot and RT-PCR; Purpose: gp100 expression in DM-6, 1520, A549

Technique: UV-Vis spectrophotometry; Purpose: Drug loading and release; coumarin-6 uptake

Technique: MTS assay; Purpose: In vitro therapeutic efficacy and cyto-compatibility

Technique: Blood clotting and hemolysis assays; Purpose: Hemo-compatibility

Technique: In vivo/ex vivo IVIS imaging; Purpose: Biodistribution and tumor retention

Key results: Physicochemical properties: T-MNPs (1:2) were 193 ± 56 nm, PDI 0.265, zeta −36 mV; stable in saline for 48 h. Drug loading 61%; sustained trametinib release over 28 days, slowest at highest membrane ratio (1:2). - TCR confirmation: TCR β chain was present on 19LF6 cells and T-MNPs by flow cytometry. Binding to gp100-MHC was dose-dependent and higher with higher membrane ratios; T-MNPs bound significantly more than D-MNPs. - Cellular uptake: T-MNPs showed significantly higher uptake in gp100-positive DM-6 and 1520 cells compared with D-MNPs; no selectivity in gp100-negative A549 cells. Uptake increased with concentration and membrane ratio. - Therapeutic efficacy: IC50 of free trametinib: 29.3 µg/mL (1520) and 1.66 µg/mL (DM-6). Trametinib-loaded T-MNPs produced significantly greater cell death than free drug, NNPs, A-MNPs, and D-MNPs at IC50 and IC75 concentrations. - Compatibility: T-MNPs were cyto-compatible up to 1000 µg/mL on HDF cells; hemolysis <5% up to 500 µg/mL; no significant effect on blood clotting up to 1000 µg/mL. - In vivo biodistribution: T-MNPs achieved more than twofold higher tumor retention than D-MNPs and NNPs, with accumulation within 6 h and retention at 24 h. Ex vivo imaging showed less liver accumulation for T-MNPs compared with controls.

Interpretation: The authors conclude that T-cell membrane-camouflaged PLGA nanoparticles expressing an anti-gp100/HLA-A2 TCR provide a biomimetic, melanoma-specific theragnostic platform with enhanced targeting, sustained drug release, and improved tumor retention. This approach may overcome limitations of conventional chemotherapy and cell-based immunotherapy for melanoma.

Limitations: No in vivo therapeutic efficacy data: The study only reports biodistribution and tumor retention, not tumor growth inhibition or survival benefit in treated mice. - No long-term toxicity or immune response evaluation: The authors note that cell membrane proteins have unknown host interactions and require further immune/toxicity profiling. - Membrane isolation and scalability: Cell membrane isolation is not robust, is labor-intensive, and requires quality control for clinical translation. - Limited animal model: Only a subcutaneous xenograft in athymic nude mice; no immunocompetent model, no spontaneous or metastatic melanoma model. - No comparison with standard-of-care melanoma therapies or combination with immune checkpoint inhibitors. - Small sample size for in vivo biodistribution (n = 6/group) and no survival endpoint.

Biochemical Pharmacology*, Volume 173, 2020, Article 113648 DOI/

7-Ketocholesterol and 7β-Hydroxycholesterol — In Vitro and Animal Models, Biological Activities, and Toxicity-Preventing Molecules

Nature Reviews Drug Discovery

Engineering Precision Nanoparticles for Drug Delivery

Materials* (MDPI; inferred from format)

Generation of Well-Defined Micro/Nanoparticles via Advanced Manufacturing Techniques for Therapeutic Delivery

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