Efficient Delivery of Tyrosinase Related Protein‐2 (TRP2) Peptides to Lymph Nodes using Serum‐Derived Exosomes

Abstract

Exosomes (EXO) are considered to be versatile carriers for biomolecules; however, the delivery of therapeutic peptides using EXOs poses several challenges. In this study, the efficiency of serum‐derived EXOs in delivering tyrosinase‐related protein‐2 (TRP2) peptides to lymph nodes is determined. TRP2 peptides are successfully incorporated into EXOs, which show a uniform and narrow size distribution of around 45 nm. The TRP2‐incorporated exosomes (EXO‐TRP2) are efficiently internalized into macrophages and dendritic cells, and are seen to display a punctate distribution. EXOs loaded with TRP2 together with MPLA, (EXO‐MPLA‐TRP2) result in a strong release of proinflammatory cytokines (TNF‐α and IL‐6) from both RAW264.7 and DC2.4 cells. Finally, subcutaneous injection of fluorescently labeled EXO‐TRP2 followed by ex vivo imaging using in vivo imaging system (IVIS) show a strong fluorescent signal in the lymph nodes after only 1 h, which is maintained until at least 4 h after injection. Taken together, the findings suggest that serum‐derived EXOs can serve as promising carriers to deliver therapeutic peptides to lymph nodes for immunotherapy.

Mannose‐Modified Serum Exosomes for the Elevated Uptake to Murine Dendritic Cells and Lymphatic Accumulation

“…While MPLA incorporated EXO‐PEG‐man greatly boosted releases of pro‐inflammatory cytokines, free EXO‐PEG‐man treated DCs exhibited negligible amount of TNF‐α and IL‐6 (Figure S5, Supporting Information). In our previous study, free MPLA showed reduced cytokine induction compared to EXO‐MPLA . As expected, EXO‐PEG‐man showed the highest pro‐inflammatory cytokine induction for the DC2.4 cells.…”

“…To examine whether the enhanced intracellular uptake of EXO‐PEG‐man into the DC2.4 cells leads to an elevated cytokine release, the pro‐inflammatory cytokines were quantitatively measured after the treatment of EXO‐PEG‐man into the DC2.4 cells. As an immune stimulator, MPLA was incorporated into EXO‐PEG‐man . After MPLA was incorporated into EXO‐PEG‐man via co‐incubation, the MPLA embedded EXO‐PEG‐man was treated to DC2.4 cells and the released cytokines (TNF‐α and IL‐6) were measured by ELISA.…”

“…Accordingly, nanosized carrier systems have been vigorously developed for lymph node targeted delivery of adjuvants and antigens . In our previous studies, serum‐derived EXOs showed an excellent lymphatic accumulation after an intradermal injection, which allowed excellent delivery of antigen peptides and adjuvants and elicited efficient T‐cell activation . Exosomes (EXOs) are extracellular vesicles consisting of a lipid bilayer with various biological components (e.g., proteins, DNA, and RNA) originating from most cells .…”

Exosomal long non-coding RNAs: biological properties and therapeutic potential in cancer treatment

“…These advantages make them ideal biomarkers for disease diagnosis and prognosis, and a promising choice as drug delivery vehicles. For example, serum-derived exosomes have been used to deliver tyrosinaserelated protein-2 (TRP2) peptides to lymph nodes in preclinical trials (Park et al, 2018). However, the application of exosomes in clinical settings remains LncRNA-SNHG14 Breast cancer Promotes trastuzumab chemoresistance Dong et al, 2018 challenging because they are currently expensive to isolate, difficult to detect and classify due to heterogeneity, and show discrepancies between in vitro and in vivo experimental systems (Li Q et al, 2018).…”

Citraconylated exosomes for improved internalization into macrophages

“…Recently, biologically derived vesicles, e.g., cell-derived vesicles and microvesicles, have been intensively investigated as one of the alternatives for synthetic particulate systems. In particular, exosomes (EXOs) have been considered promising drug carriers because of their aqueous stability, homogeneous diameter, and biocompatibility [11][12][13]. In a previous study, serum derived EXOs had a narrow size distribution of approximately 50 nm as well as high production yield [14].…”