Canine Mesenchymal-Stem-Cell-Derived Extracellular Vesicles Attenuate Atopic Dermatitis

Abstract

Atopic dermatitis (AD) is a chronic inflammatory skin disease that is associated with systemic inflammation and immune modulation. Previously, we have shown that extracellular vesicles resulting from human adipose-tissue-derived mesenchymal stem cells (ASC-EVs) attenuated AD-like symptoms by reducing the levels of multiple inflammatory cytokines. Here, we aimed to investigate the improvement of canine AD upon using canine ASC-exosomes in a Biostir-induced AD mouse model. Additionally, we conducted in vivo toxicity studies to determine whether they targeted organs and their potential toxicity. Firstly, we isolated canine ASCs (cASCs) from the adipose tissue of a canine and characterized the cASCs-EVs. Interestingly, we found that cASC-EVs improved AD-like dermatitis and markedly decreased the levels of serum IgE, ear thickness, inflammatory cytokines, and chemokines such as IL-4 and IFN-γ in a dose-dependent manner. Moreover, there was no systemic toxicity in single- or repeat-dose toxicity studies using ICR mice. In addition, we analyzed miRNA arrays from cASC-EVs using next-generation sequencing (NGS) to investigate the role of miRNAs in improving inflammatory responses. Collectively, our results suggest that cASC-EVs effectively attenuate AD by transporting anti-inflammatory miRNAs to atopic lesions alongside no toxicological findings, resulting in a promising cell-free therapeutic option for treating canine AD.

Beyond Canine Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells Transplantation: An Update on Their Secretome Characterization and Applications

“…In a more recent study using the same isolation method, the EV size was around 104 nm, they were positive for CD81, and had lower levels of calnexin compared to the cell lysate, which confirmed their identity [36]. Additionally, 798 miRNAs in AT-MSC EVs were identified using next-generation sequencing (NGS).…”

“…Additionally, 798 miRNAs in AT-MSC EVs were identified using next-generation sequencing (NGS). Some of the top seven miR-NAs (let-7a, let-7b, miR-21, let-7f, miR-125b, miR-24, and miR-29a), accounting for 66% of the total miRNAs, are known to be involved in the regulation of inflammation [36]. Particularly, let-7a, let-7b, and miR-21 have anti-inflammatory actions through miRNA transfer into the recipient cells [37][38][39].…”

“…Particularly, let-7a, let-7b, and miR-21 have anti-inflammatory actions through miRNA transfer into the recipient cells [37][38][39]. According to the Gene Ontology (GO) analysis of the top 20 target miRNAs, they were related to the intrinsic apoptotic signaling pathway in response to DNA damage (biological process), protein serine/threonine kinase activity (molecular function), and the JAK-STAT signaling pathway [36]. Serine/threonine kinase play key roles in cellular growth, differentiation, and secondary metabolism by sensing changes in the external environment and altering the gene expression to trigger the proper metabolic processes [40].…”

Canine Mesenchymal Stromal Cell Exosomes: State-of-the-Art Characterization, Functional Analysis and Applications in Various Diseases

“…Additionally, this method can be employed to detect the presence of antigens by the use of fluorescently labeled antibodies [77]. This method is most commonly used to characterize canine MSC exosomes, as supported by the previously reported literature [40,78].…”

“…To this end, many scientists have reported methods to isolate MSCs from the above sources. Briefly, adipose tissue/bone marrow are isolated from healthy animals and cultured in vitro in the presence of FBS [9,40]. Once cells reach the desired confluence, culture supernatants are extracted for exosome isolation.…”