Nanotechnological engineering of extracellular vesicles for the development of actively targeted hybrid nanodevices

Abstract

Background We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeutic stimuli-responsive cargo and equipping them with antibodies for efficient targeting of lymphoma cells. Results The post-isolation engineering of the EVs is accomplished by a freeze–thaw method to load therapeutically-active zinc oxide nanocrystals (ZnO NCs), obtaining the so-called TrojanNanoHorse (TNH) to recall the biomimetism and cytotoxic potential of this novel nanoconstruct. TNHs are further modified at their surface with anti-CD20 monoclonal antibodies (TNHCD20) achieving specific targeting against lymphoid cancer cell line. The in vitro characterization is carried out on CD20+ lymphoid Daudi cell line, CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (B lymphocytes). The TNH shows nanosized structure, high colloidal stability, even over time, and good hemocompatibility. The in vitro characterization shows the high biocompatibility, targeting specificity and cytotoxic capability. Importantly, the selectivity of TNHCD20 demonstrates significantly higher interaction towards the target lymphoid Daudi cell line compared to the CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (lymphocytes). An enhanced cytotoxicity directed against Daudi cancer cells is demonstrated after the TNHCD20 activation with high-energy ultrasound shock-waves (SW). Conclusion This work demonstrates the efficient re-engineering of EVs, derived from healthy cells, with inorganic nanoparticles and monoclonal antibodies. The obtained hybrid nanoconstructs can be on-demand activated by an external stimulation, here acoustic pressure waves, to exploit a cytotoxic effect conveyed by the ZnO NCs cargo against selected cancer cells. Graphical Abstract

Stimuli-responsive hybrid nanoconstructs for efficient theranostic applications in nanomedicine

“…The SWs alone caused a slight decrease in the cell proliferation in adenocarcinoma KB cell line, while the combination with pure ZnO NCs showed an important cytotoxicity. The same result was then reported with the final TNH CD20 nanocostruct, showing remarkable and selective toxicity with SWs directed to leukaemic cancer cells with respect to the healthy lymphocytes (Dumontel, 2022). The observed cytotoxicity could be caused by the combination of various concurrent effects leading to the mechanical injury of the cell structure: (i) an enhanced acoustic bubble cavitation;…”

“…Furthermore, the anti CD20 antibody was bioconjugated to the EV surface, creating the final TNH CD20 nanoconstruct. The monodispersed size distribution, the colloidal stability over long time periods of the TNH, its haemocompatibility and biocompatibility were then all successfully assessed; the targeting selectivity of the TNH CD20 towards Daudi cancer cells, sparing healthy B lymphocytes (also CD20 + ) and cancer HL60 cells (which are CD20negative cells) was also efficaciously demonstrated (Dumontel, 2022). With the TNH CD20 weapon prepared against leukaemia cancer cells, we then studied the therapeutic and bioimaging capabilities.…”

“…Concerning the therapeutic activity, the synergism between the TNH CD20 and the applied shock waves (SWs), a specific type of mechanical pressure waves, was proven to kill Daudi cancer cells and spare healthy B lymphocytes, in vitro (Dumontel et al, 2022) (Figure 1B).…”

Biomimicking Extracellular Vesicles with Fully Artificial Ones: A Rational Design of EV-BIOMIMETICS toward Effective Theranostic Tools in Nanomedicine

“…It is then foreseen to load the artificial EVs with active compounds, enabling an advanced function in terms of imaging reporter, diagnostic, or therapeutic activities. Examples of these various functions can derive from the already-reported literature concerning both natural EVs or liposomes, proposing the incorporation of drugs, , genetic materials, organic dyes, aptamers, peptides, and more in general proteins ranging from enzyme to antibodies, and even solid-state nanoparticles of various nature, size, porosities, properties, and functions. − …”

Exploring the mechanisms of drug-delivery by decorated ZnO nanoparticles through predictive ReaxFF molecular dynamics simulations

“…8 Zinc Oxide (ZnO) nanoparticles (NPs) are very promising for anticancer applications because they can induce the generation of reactive oxygen species (ROS) and thus oxidative stress that leads to antimicrobial activity. 9 Recently, an innovative hybrid nanosystem including stimuli-responsive ZnO nanoparticles coated with extracellular vesicles (EVs) equipped with antibodies has been engineered by Dumontel and coworkers 10 to reach the target cells more effectively. This nano construct improved the therapeutic effects, 11 relying on the combined actions of ZnONPs and the monoclonal antibody carried on the EV shell.…”