Extracellular vesicle-based drug overview: research landscape, quality control and nonclinical evaluation strategies

Gangling Xu, Jing Jin, Zhihao Fu and 4 more (opens in a new tab)

Signal Transduction and Targeted Therapy · 2025 · Review · CC BY

doi:10.1038/s41392-025-02312-w (opens in a new tab)

Abstract

Extracellular vesicles share lipid‒protein membranes with their parent cells, allowing for the targeted transfer of bioactive cargo to recipient cells for functional modulation. The biological features allow extracellular vesicles to serve both as intrinsic therapeutics and as engineered delivery vehicles for targeted molecule transport. In recent years, extracellular vesicle-based therapy has shown great potential as a new therapeutic approach for traumatic conditions and degenerative, acute, and refractory diseases. As extracellular vesicle engineering continues to evolve, more innovative drugs are expected to receive investigational new drug approvals and marketing approvals from regulatory agencies in the future. However, many challenges exist in terms of mechanistic understanding, engineering modifications, manufacturing processes, quality control, and nonclinical research, and no drug regulatory authorities have currently issued specific technical evaluation guidelines for extracellular vesicle-based drugs, all of which have hindered the clinical translation of these drugs. In this article, which is focused primarily on extracellular vesicles derived from mammalian cells, we summarize the clinical translation and process development research status of extracellular vesicle-based drugs and propose both general considerations and key aspects of quality control strategies and nonclinical evaluations in the development process. The aim of this review is to provide valuable references for the development and evaluation of extracellular vesicle-based products, accelerate the clinical translation process, and benefit patients as soon as possible.

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Lewis ND, et al. Exosome surface display of IL12 results in tumor-retained pharmacology with superior potency and limited systemic exposure compared with recombinant IL12. Mol. Cancer Ther. 2021;20:523-534.
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115.
ExoSTING, an extracellular vesicle loaded with STING agonists, promotes tumor immune surveillance · 2021
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Zhou Y, et al. Tumor-specific delivery of KRAS siRNA with iRGD-exosomes efficiently inhibits tumor growth. ExRNA. 2019;1:28.
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117.
Bai J, et al. Engineered targeting tLyp-1 exosomes as gene therapy vectors for efficient delivery of siRNA into lung cancer cells. Asian J. Pharm. Sci. 2020;15:461-471.
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118.
Liang Y, et al. Chondrocyte-targeted MicroRNA delivery by engineered exosomes toward a cell-free osteoarthritis therapy. ACS Appl Mater. Interfaces. 2020;12:36938-36947.
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Villarroya-Beltri C, et al. Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs. Nat. Commun. 2013;4:2980.
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Epigenetic reprogramming enhances the therapeutic efficacy of osteoblast‐derived extracellular vesicles to promote human bone marrow stem cell osteogenic differentiation · 2021
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122.
Man, K. et al. Epigenetic reprogramming via synergistic hypomethylation and hypoxia enhances the therapeutic efficacy of mesenchymal stem cell extracellular vesicles for bone repair. Int. J. Mol. Sci. 24, 7564 (2023).
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Incorporation of small extracellular vesicles in sodium alginate hydrogel as a novel therapeutic strategy for myocardial infarction · 2019
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127.
Injectable exosome-functionalized extracellular matrix hydrogel for metabolism balance and pyroptosis regulation in intervertebral disc degeneration · 2021
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128.
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Kulkarni JA, et al. Design of lipid nanoparticles for in vitro and in vivo delivery of plasmid DNA. Nanomedicine. 2017;13:1377-1387.
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Zhuang M, et al. SPION decorated exosome delivery of TNF-alpha to cancer cell membranes through magnetism. Nanoscale. 2020;12:173-188.
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136.
Yu T, et al. Extracellular vesicle-functionalized bioactive scaffolds for bone regeneration. Asian J. Pharm. Sci. 2024;19:100945.
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137.
Self-Healing Hydrogels: Preparation, Mechanism and Advancement in Biomedical Applications · 2021
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138.
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Jiang T, et al. ADSC-exo@MMP-PEG smart hydrogel promotes diabetic wound healing by optimizing cellular functions and relieving oxidative stress. Mater. Today Bio. 2022;16:100365.
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141.
Han Y, et al. Biomimetic injectable hydrogel microspheres with enhanced lubrication and controllable drug release for the treatment of osteoarthritis. Bioact. Mater. 2021;6:3596-3607.
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142.
Microneedles in Drug Delivery: Progress and Challenges · 2021
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Wan T, et al. Exosome-mediated delivery of Cas9 ribonucleoprotein complexes for tissue-specific gene therapy of liver diseases. Sci. Adv. 2022;8:eabp9435.
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150.
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151.
Sun L, et al. Menstrual blood-derived stem cells exosomal miR-let-7 to ameliorate pulmonary fibrosis through inhibiting ferroptosis by Sp3/HDAC2/Nrf2 signaling pathway. Int. Immunopharmacol. 2024;126:111316.
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152.
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Hayon Y, et al. Platelet microparticles induce angiogenesis and neurogenesis after cerebral ischemia. Curr. Neurovasc. Res. 2012;9:185-192.
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155.
Secretomes of apoptotic mononuclear cells ameliorate neurological damage in rats with focal ischemia [v2; ref status: indexed, http://f1000r.es/4kv] · 2014
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156.
Doeppner TR, et al. Extracellular vesicles improve post-stroke neuroregeneration and prevent postischemic immunosuppression. Stem Cells Transl. Med. 2015;4:1131-1143.
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Zhang Y, et al. Exosomes derived from mesenchymal stromal cells promote axonal growth of cortical neurons. Mol. Neurobiol. 2017;54:2659-2673.
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158.
Secondary Release of Exosomes from Astrocytes Contributes to the Increase in Neural Plasticity and Improvement of Functional Recovery after Stroke in Rats Treated with Exosomes Harvested from MicroRNA 133b-Overexpressing Multipotent Mesenchymal Stromal Cells · 2017
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159.
Yang Y, et al. Exosomes secreted by adipose-derived stem cells contribute to angiogenesis of brain microvascular endothelial cells following oxygen-glucose deprivation in vitro through MicroRNA-181b/TRPM7 axis. J. Mol. Neurosci. 2018;65:74-83.
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160.
Daly T, et al. Amyloid-н╡ in Alzheimer’s disease: a study of citation practices of the amyloid cascade hypothesis between 1992 and 2019. J. Alzheimers Dis. 2020;74:1309-1317.
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Yuyama K, et al. Decreased amyloid-н╡ pathologies by intracerebral loading of glycosphingolipid-enriched exosomes in Alzheimer model mice. J. Biol. Chem. 2014;289:24488-24498.
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Cui GH, et al. Exosomes derived from hypoxia-preconditioned mesenchymal stromal cells ameliorate cognitive decline by rescuing synaptic dysfunction and regulating inflammatory responses in APP/PS1 mice. FASEB J. 2018;32:654-668.
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163.
Fe65-engineered neuronal exosomes encapsulating corynoxine-B ameliorate cognition and pathology of Alzheimer’s disease · 2023
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164.
Mesenchymal stem cell-derived exosomes promote neurogenesis and cognitive function recovery in a mouse model of Alzheimer’s disease · 2019
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165.
Lee M, et al. The exosome of adipose-derived stem cells reduces н╡-amyloid pathology and apoptosis of neuronal cells derived from the transgenic mouse model of Alzheimer’s disease. Brain Res. 2018;1691:87-93.
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166.
RVG-modified exosomes derived from mesenchymal stem cells rescue memory deficits by regulating inflammatory responses in a mouse model of Alzheimer’s disease · 2019
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167.
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Mesenchymal Stem/Stromal Cell-Derived Extracellular Vesicles and Their Potential as Novel Immunomodulatory Therapeutic Agents · 2017
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169.
Shokravi S, et al. Mesenchymal stromal cells (MSCs) and their exosome in acute liver failure (ALF): a comprehensive review. Stem Cell Res Ther. 2022;13:192.
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170.
Liu Y, et al. AMSC-derived exosomes alleviate lipopolysaccharide/d-galactosamine-induced acute liver failure by miR-17-mediated reduction of TXNIP/NLRP3 inflammasome activation in macrophages. Ebiomedicine. 2018;36:140-150.
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171.
Zhao S, Liu Y, Pu Z. Bone marrow mesenchymal stem cell-derived exosomes attenuate D-GaIN/LPS-induced hepatocyte apoptosis by activating autophagy in vitro. Drug Des. Dev. Ther. 2019;13:2887-2897.
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172.
Wu HY, et al. Exosomes derived from human umbilical cord mesenchymal stem cells alleviate acetaminophen-induced acute liver failure through activating ERK and IGF-1R/PI3K/AKT signaling pathway. J. Pharm. Sci. 2021;147:143-155.
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173.
Hu Z, et al. MSCs Deliver Hypoxia-treated mitochondria reprogramming acinar metabolism to alleviate severe acute pancreatitis injury. Adv. Sci. 2023;10:e2207691.
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174.
Ma Z, et al. Exosomes from TNF-α preconditioned human umbilical cord mesenchymal stromal cells inhibit the autophagy of acinar cells of severe acute pancreatitis via shuttling bioactive metabolites. Cell Mol. Life Sci. 2023;80:257.
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175.
Ren S, et al. miR-29a-3p transferred by mesenchymal stem cells-derived extracellular vesicles protects against myocardial injury after severe acute pancreatitis. Life Sci. 2021;272:119189.
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176.
Chen M, et al. Exosomes from human induced pluripotent stem cells derived mesenchymal stem cells improved myocardial injury caused by severe acute pancreatitis through activating Akt/Nrf2/HO-1 axis. Cell Cycle. 2022;21:1578-1589.
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177.
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178.
Zheng G, et al. Mesenchymal stromal cell-derived extracellular vesicles: regenerative and immunomodulatory effects and potential applications in sepsis. Cell Tissue Res. 2018;374:1-15.
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179.
CAR exosomes derived from effector CAR-T cells have potent antitumour effects and low toxicity · 2019
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180.
Zhang X, et al. Engineered extracellular vesicles for cancer therapy. Adv. Mater. 2021;33:e2005709.
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181.
Engineered exosomes from different sources for cancer-targeted therapy · 2023
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182.
Kugeratski, F. G., McAndrews, K. M. & Kalluri, R. Multifunctional applications of engineered extracellular vesicles in the treatment of cancer. Endocrinology. 162, bqaa250 (2021).
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Zhou X, Jia Y, Mao C, Liu S. Small extracellular vesicles: Non-negligible vesicles in tumor progression, diagnosis, and therapy. Cancer Lett. 2024;580:216481.
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184.
Yu S, et al. Research progress and application of the CRISPR/Cas9 gene-editing technology based on hepatocellular carcinoma. Asian J. Pharm. Sci. 2023;18:100828.
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185.
PD-1/PD-L1 checkpoint inhibitors in advanced hepatocellular carcinoma immunotherapy · 2022
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186.
Nebulized exosomes derived from allogenic adipose tissue mesenchymal stromal cells in patients with severe COVID-19: a pilot study · 2022
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187.
Nebulization Therapy with Umbilical Cord Mesenchymal Stem Cell-Derived Exosomes for COVID-19 Pneumonia · 2022
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188.
Lightner AL, et al. Bone marrow mesenchymal stem cell-derived extracellular vesicle infusion for the treatment of respiratory failure from COVID-19: A randomized, placebo-controlled dosing clinical trial. Chest. 2023;164:1444-1453.
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Green, O. et al. Inhaled exosomes genetically manipulated to overexpress CD24 (EXO-CD24) as a compassionate use in severe ARDS patients. Biomedicines.11, 2523 (2023).
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191.
Safety of intraparenchymal injection of allogenic placenta mesenchymal stem cells derived exosome in patients undergoing decompressive craniectomy following malignant middle cerebral artery infarct, a pilot randomized clinical trial · 2022
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192.
Xie, X. et al. Clinical safety and efficacy of allogenic human adipose mesenchymal stromal cells-derived exosomes in patients with mild to moderate Alzheimer’s disease: a phase I/II clinical trial. Gen. Psychiatr. 36, e101143 (2023).
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Elucidating Methods for Isolation and Quantification of Exosomes: A Review · 2021
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Tangential Flow Filtration for Highly Efficient Concentration of Extracellular Vesicles from Large Volumes of Fluid · 2018
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202.
Scalable, cGMP‐compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL‐15/lactadherin complexes · 2018
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203.
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Anticoli S, et al. Engineered exosomes emerging from muscle cells break immune tolerance to HER2 in transgenic mice and induce antigen-specific CTLs upon challenge by human dendritic cells. J. Mol. Med. 2018;96:211-221.
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218.
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Hadiwinoto GD, Lip Kwok PC, Lakerveld R. A review on recent technologies for the manufacture of pulmonary drugs. Ther. Deliv. 2018;9:47-70.
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Mesenchymal stem cells reverse EMT process through blocking the activation of NF-κB and Hedgehog pathways in LPS-induced acute lung injury · 2020
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Sui X, Liu W, Liu Z. Exosomal lncRNA-p21 derived from mesenchymal stem cells protects epithelial cells during LPS-induced acute lung injury by sponging miR-181. Acta Biochim Biophys. Sin. 2021;53:748-757.
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Prospects and challenges of extracellular vesicle-based drug delivery system: considering cell source · 2020
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230.
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