In Vivo Differentiation of Therapeutic Insulin-Producing Cells from Bone Marrow Cells via Extracellular Vesicle-Mimetic Nanovesicles

Abstract

The current diabetes mellitus pandemic constitutes an important global health problem. Reductions in the mass and function of β-cells contribute to most of the pathophysiology underlying diabetes. Thus, physiological control of blood glucose levels can be adequately restored by replacing functioning β-cell mass. Sources of functional islets for transplantation are limited, resulting in great interest in the development of alternate sources, and recent progress regarding cell fate change via utilization of extracellular vesicles, also known as exosomes and microvesicles, is notable. Thus, this study investigated the therapeutic capacity of extracellular vesicle-mimetic nanovesicles (NVs) derived from a murine pancreatic β-cell line. To differentiate insulin-producing cells effectively, a three-dimensional in vivo microenvironment was constructed in which extracellular vesicle-mimetic NVs were applied to subcutaneous Matrigel platforms containing bone marrow (BM) cells in diabetic immunocompromised mice. Long-term control of glucose levels was achieved over 60 days, and differentiation of donor BM cells into insulin-producing cells in the subcutaneous Matrigel platforms, which were composed of islet-like cell clusters with extensive capillary networks, was confirmed along with the expression of key pancreatic β-cell markers. The resectioning of the subcutaneous Matrigel platforms caused a rebound in blood glucose levels and confirmed the source of functioning β-cells. Thus, efficient differentiation of therapeutic insulin-producing cells was attained in vivo through the use of extracellular vesicle-mimetic NVs, which maintained physiological glucose levels.

Modularized Extracellular Vesicles: The Dawn of Prospective Personalized and Precision Medicine

“…Although EVMs exhibit characteristics extremely similar to Exos,53, 54, 124 the actual identity of EVMs needs to be clarified by further investigation.…”

“…Furthermore, it has gradually been revealed that EVMs show similar properties to EVs;53, 54 therefore, the method of producing EVMs might be a much more effective way to manufacture modularized EVs.…”

Engineering Extracellular Vesicles as Nanotherapeutics for Regenerative Medicine

“…Regardless, functional studies showed that the nanovesicles directed bone marrow cell differentiation into insulin-producing cells in vitro, that when transplanted into diabetic mice, were capable of controlling blood glucose levels for 60 days. Though no control comparison was made to native EVs derived from the same β-cell line, a yield of 8.2 × 10 8 particles per 1 µg of total protein was reported, confirming that generally a high number of vesicles can be made from this method [105]. A similar protocol was used to produce EV mimics from human breast epithelial cells [106].…”

“…As such, it is especially remarkable when authors showed that synthetic EVs were able to be produced at about a 100-fold greater yield than native EVs secreted by the same number of cultured cells. Similar methods were used to synthesize EV-mimicking nanovesicles from pancreatic β-cells in which cells were harvested and serially extruded through 10, 5 and 1 μm pore-sized polycarbonate membrane filters [105]. Assessment of the physical properties of nanovesicles revealed an average size of 203 nm, and it has been found that protein contents similar to donor cells though insulin and Pdx1, a transcription factor that regulates β-cell development, were not detected.…”

Targeting Inflammatory Vasculature by Extracellular Vesicles

“…Based on this concept, siRNA or miRNA could be loaded in cells, and then, the RNA would be capsulated in EVs during their formation in culture media. Another loading strategy is to mix or incubate drugs with cells, and drug-loaded vesicles could be obtained by extrusion of cells (47) (48) (42). Jang’s group (47) produced doxorubicin-loaded nanovesicles by serial extrusions of cells via a membrane filter, and drug-loaded vesicles were purified using a series of density gradient centrifugations.…”