Tumor-targeted multifunctional extracellular vesicles as drug carriers for lung cancer therapy

Abstract

Aim: Chemotherapy continues to be the frontline treatment for lung cancer patients. However, treatment-related toxicity and off-target effects limit the use of chemotherapy. Therefore, improvements in delivering chemotherapeutics with reduced toxicity to normal tissues are needed. In the present study, we combined nanotechnology with extracellular vesicle (EV) technology to produce tumor-targeted multifunctional EVs (tt-Mfn-EVs) as drug carriers for cancer therapy.

Methods: The tt-Mfn-EVs were formulated by exogenously loading EVs with gold nanoparticles conjugated to cisplatin (CDDP) via pH-sensitive coordination ester linkage. Attached to the outer surface of drug-loaded EVs is the transferrin ligand for targeting transferrin receptor (TfR) overexpressing lung cancer cells.

Results: The tt-Mfn-EVs were 138.2 nm in size and exhibited greater drug release kinetics at pH 5.5 compared to pH 7.2. They significantly reduced cell viability of A₅₄₉ (TfR high) lung cancer cells compared to HCC₈₂₇ (TfR low) cells and non-targeted EVs. Tt-Mfn-EVs also induced higher levels of apoptosis and DNA damage in A₅₄₉ and HCC₈₂₇ cells compared to control groups. Finally, tt-Mfn-EV-mediated cytotoxicity was minimal in normal human lung fibroblast (MRC-₉) and human embryonic kidney 293 (HEK₂₉₃) cells compared to free CDDP.

Conclusion: Our study showed that tt-Mfn-EVs exerted selective and enhanced tumor-targeted cell killing in vitro, providing an opportunity for developing EV-based drug carriers for cancer therapy.

Keywords

Extracellular vesicles / lung cancer / drug delivery / chemotherapy / tumor targeting

References

[1] Behranvand N,Zolfaghari Emameh R.Chemotherapy: a double-edged sword in cancer treatment.Cancer Immunol Immunother2022;71:507-26 PMCID:PMC10992618

[2] Gridelli C,Mok T.Face to face among different chemo-immunotherapy combinations in the first line treatment of patients with advanced non-small cell lung cancer: results of an international expert panel meeting by the italian association of thoracic oncology (AIOT).Lung Cancer2024;187:107441

[3] Sordo-Bahamonde C,Gonzalez-Rodriguez AP.Chemo-immunotherapy: a new trend in cancer treatment.Cancers2023;15:2912 PMCID:PMC10252089

[4] Hendriks LEL,Faivre-Finn C.Non-small-cell lung cancer.Nat Rev Dis Primers2024;10:71

[5] Kuderer NM,Lustberg MB.Mitigating acute chemotherapy-associated adverse events in patients with cancer.Nat Rev Clin Oncol2022;19:681-97

[6] Yazbeck V,Myers J,Cuttino L.An overview of chemotoxicity and radiation toxicity in cancer therapy.Adv Cancer Res2022;155:1-27

[7] Ioele G,Occhiuzzi MA.Anticancer drugs: recent strategies to improve stability profile, pharmacokinetic and pharmacodynamic properties.Molecules2022;27:5436 PMCID:PMC9457551

[8] Rezakhani L,Rostaminasab G.Exosomes: special nano-therapeutic carrier for cancers, overview on anticancer drugs.Med Oncol2022;40:31

[9] Ulldemolins A,Andrade F.Perspectives of nano-carrier drug delivery systems to overcome cancer drug resistance in the clinics.Cancer Drug Resist2021;4:44-68 PMCID:PMC9019183

[10] Wang B,Teng Y.Current advance of nanotechnology in diagnosis and treatment for malignant tumors.Signal Transduct Target Ther2024;9:200 PMCID:PMC11323968

[11] Sen S,Kumar N,Ranjan OP.Exosomes as natural nanocarrier-based drug delivery system: recent insights and future perspectives.3 Biotech2023;13:101 PMCID:PMC9970142

[12] Brezgin S,Yudaeva A.Basic guide for approaching drug delivery with extracellular vesicles.Int J Mol Sci2024;25:10401 PMCID:PMC11476574

[13] Liu X,Wang S,Hao H.Extracellular vesicles: powerful candidates in nano-drug delivery systems.Drug Deliv Transl Res2024;14:295-311

[14] Vader P,Pasterkamp G.Extracellular vesicles for drug delivery.Adv Drug Deliv Rev2016;106:148-56

[15] Samanta S,Drosos N,Dawn B.Exosomes: new molecular targets of diseases.Acta Pharmacol Sin2018;39:501-13 PMCID:PMC5888687

[16] Sutaria DS,Phelps MA.Achieving the promise of therapeutic extracellular vesicles: the devil is in details of therapeutic loading.Pharm Res2017;34:1053-66 PMCID:PMC5565485

[17] Batrakova EV.Using exosomes, naturally-equipped nanocarriers, for drug delivery.J Control Release2015;219:396-405 PMCID:PMC4656109

[18] Chen C,Sun M.Phospholipid-anchored ligand conjugation on extracellular vesicles for enhanced cancer targeting.Small2024;20:e2310712

[19] Herrmann IK,Fuhrmann G.Extracellular vesicles as a next-generation drug delivery platform.Nat Nanotechnol2021;16:748-59

[20] Zhang M,Liu L.Engineered exosomes from different sources for cancer-targeted therapy.Signal Transduct Target Ther2023;8:124 PMCID:PMC10017761

[21] Antimisiaris SG,Marazioti A.Exosomes and exosome-inspired vesicles for targeted drug delivery.Pharmaceutics2018;10:218 PMCID:PMC6321407

[22] Familtseva A,Tyagi SC.Exosomes: cell-created drug delivery systems.Mol Cell Biochem2019;459:1-6

[23] Jaiswal N,Chanda N.Nanogold-albumin conjugates: transformative approaches for next-generation cancer therapy and diagnostics.Nanoscale2025;17:11191-220

[24] Lee JW,Heo JH.Simultaneous stabilization and functionalization of gold nanoparticles via biomolecule conjugation: progress and perspectives.ACS Appl Mater Interfaces2021;13:42311-28

[25] Kesharwani P,Sang L.Gold nanoparticles and gold nanorods in the landscape of cancer therapy.Mol Cancer2023;22:98 PMCID:PMC10283337

[26] Kim Y,Shin H.Advanced precision dual photothermal and photodynamic therapy for prostate cancer using PSMA-ICG-conjugated gold nanorods.ACS Biomater Sci Eng2025;11:3376-88

[27] Zhao T,Shi J.Engineering the protein corona: strategies, effects, and future directions in nanoparticle therapeutics.Biomed Pharmacother2024;175:116627

[28] Bolaños K,Jara-Guajardo P.In vivo targeted delivery of extracellular vesicle‒gold nanorod hybrids to metastatic melanoma lung tumors.Int J Nanomedicine2025;20:6983-98 PMCID:PMC12132528

[29] Srinivasarao M.Ligand-targeted drug delivery.Chem Rev2017;117:12133-64

[30] Sundaram V.Emerging biomimetic drug delivery nanoparticles inspired by extracellular vesicles.Wiley Interdiscip Rev Nanomed Nanobiotechnol2025;17:e70025 PMCID:PMC12370395

[31] Johnson V,Kumar US.Surface-engineered extracellular vesicles in cancer immunotherapy.Cancers2023;15:2838 PMCID:PMC10216164

[32] Amreddy N,Babu A.Tumor-targeted and pH-controlled delivery of doxorubicin using gold nanorods for lung cancer therapy.Int J Nanomedicine2015;10:6773-88 PMCID:PMC4631428

[33] Lan X,Shao L.Effect of transferrin-modified Fe₃O₄ nanoparticle targeted delivery miR-15a-5p combined with photothermal therapy on lung cancer.Thorac Cancer2025;16:e15497 PMCID:PMC11729913

[34] Nguyen NT,Tran NTN.Synergic activity against MCF-7 breast cancer cell growth of nanocurcumin-encapsulated and cisplatin-complexed nanogels.Molecules2018;23:3347 PMCID:PMC6321001

[35] Zhang Z,Wen T.Quantification of cisplatin encapsulated in nanomedicine: an overview.Biosensors2025;15:293 PMCID:PMC12110417

[36] Inoue S,Branch CD.mda-7 In combination with bevacizumab treatment produces a synergistic and complete inhibitory effect on lung tumor xenograft.Mol Ther2007;15:287-94

[37] Muralidharan R,Amreddy N.Tumor-targeted nanoparticle delivery of HuR siRNA inhibits lung tumor growth in vitro and in vivo by disrupting the oncogenic activity of the RNA-binding protein HuR.Mol Cancer Ther2017;16:1470-86 PMCID:PMC5544587

[38] Welsh JA, Goberdhan DCI, O’Driscoll L, et al.; MISEV Consortium. Minimal information for studies of extracellular vesicles (MISEV2023): from basic to advanced approaches.J Extracell Vesicles2024;13:e12404 PMCID:PMC10850029

[39] Jeppesen DK,Franklin JL.Reassessment of exosome composition.Cell2019;177:428-45.e18 PMCID:PMC6664447

[40] Fathy MM,Bashandy YS.An insight into synthesis and antitumor activity of citrate and gallate stabilizing gold nanospheres.Sci Rep2023;13:2749 PMCID:PMC9935520

[41] Zhou W,Liu D.Gold nanoparticles for in vitro diagnostics.Chem Rev2015;115:10575-636 PMCID:PMC5226399

[42] Beit-Yannai E,Stamer WD.Physical exosome:exosome interactions.J Cell Mol Med2018;22:2001-6 PMCID:PMC5824382

[43] Dreyer F.Biogenesis and functions of exosomes and extracellular vesicles.Methods Mol Biol2016;1448:201-16

[44] Persi E,Damaghi M.Systems analysis of intracellular pH vulnerabilities for cancer therapy.Nat Commun2018;9:2997 PMCID:PMC6068141

[45] Sergeeva TF,Zlobovskaya OA.Relationship between intracellular pH, metabolic co-factors and caspase-3 activation in cancer cells during apoptosis.Biochim Biophys Acta Mol Cell Res2017;1864:604-11

[46] Kennedy L,Harper ME.Role of glutathione in cancer: from mechanisms to therapies.Biomolecules2020;10:1429 PMCID:PMC7600400

[47] Li Y,Lin J,Yang S.Smart nanomedicine agents for cancer, triggered by pH, glutathione, H₂O₂, or H₂S.Int J Nanomedicine2019;14:5729-49 PMCID:PMC6664425

[48] Liu G,Zhang Y.Engineering biomimetic platesomes for pH-responsive drug delivery and enhanced antitumor activity.Adv Mater2019;31:e1900795

[49] Ciccotosto G,Galluzzi L.A novel pH-sensitive probe to quantify autophagy on high throughput/content imaging platforms.BioRxiv2025;

[50] Koltai T,Harguindey S.Chapter 3 - The pH-centered paradigm in cancer. In An innovative approach to understanding and treating cancer: targeting pH. Elsevier; 2020. pp. 53-97.

[51] Manohar S.Cisplatin nephrotoxicity: a review of the literature.J Nephrol2018;31:15-25

[52] Calzolari A,Deaglio S.Transferrin receptor 2 is frequently expressed in human cancer cell lines.Blood Cells Mol Dis2007;39:82-91

[53] Liu Y,Vo-Dinh T.Gold nanoparticles-mediated photothermal therapy and immunotherapy.Immunotherapy2018;10:1175-88

[54] Zhou M,Cheng Y,Yang J.Application of gold-based nanomaterials in tumor photothermal therapy and chemotherapy.J Biomed Nanotechnol2020;16:739-62