Multivalent Engineering of Exosomes with Activatable Aptamer Probes for Specific Regulation and Monitoring of Cell Targeting

Abstract

Reconstituting and probing exosome-cell interactions is critical for elucidating exosome-related cell biology and advancing their diagnostic and therapeutic potential. We report here an exosomal engineering strategy to achieve controlled regulation of exosome-cell interactions with activatable sensing capability. The approach relies on membrane-protein directed, programmable DNA self-assembly to construct a DNA polymeric scaffold with multivalent display of structure-switchable aptamer sensing probes on exosome surfaces. The engineered exosomes exhibit enhanced cancer cell targeting ability compared to exosomes modified with monovalent aptamers. Furthermore, the anchored aptamer probes could be activated by specific membrane protein targeting, followed by structural switching to report an output fluorescence signal, thus allowing dynamic monitoring of exosome-cell interactions both in vitro and in vivo. We envision this will provide a complementary tool for specific regulation and monitoring of exosome-cell docking interactions and will advance the development of exosome-based biomedical applications.

Extracellular vesicles in renal cell carcinoma: challenges and opportunities coexist

“…Through surface modifications or the incorporation of therapeutic moieties, engineered sEVs can achieve precise delivery of therapeutics, thus enhancing their selective accumulation at the tumor site, while reducing non-specific dissemination throughout the body ( 81 ). These advancements have highlighted the enormous potential of engineered sEVs for safe, targeted, and effective drug delivery, paving the way for the development of more advanced and personalized therapies ( 82 ).…”

Magnetic-nanowaxberry-based microfluidic ExoSIC for affinity and continuous separation of circulating exosomes towards cancer diagnosis

“…Exosomes are membrane-bound lipid nanovesicles that play a vital role in intercellular communication and are typically 30–150 nm in size. 1–3 Matured exosomes can influence the behavior of recipient cells through a variety of pathways such as interactions with membrane receptors, membrane fusion or uptake by cells (macropinocytosis, phagocytosis, etc. ).…”

“…Exosomes are membrane-bound lipid nanovesicles that play a vital role in intercellular communication and are typically 30-150 nm in size. [1][2][3] Matured exosomes can influence the behavior of recipient cells through a variety of pathways such as interactions with membrane receptors, membrane fusion or uptake by cells (macropinocytosis, phagocytosis, etc.). 4,5 In this way, exosomes are involved in a wide range of physiological and pathological processes in mammals, such as immune response and disease development (metabolic diseases, cardiovascular diseases, neurodegeneration, tumors, etc.).…”

Comprehensive Detection of PD-L1 Protein and mRNA in Tumor Cells and Extracellular Vesicles through a Real-Time qPCR Assay

“…For instance, captured EVs/cells are broken by heating, and the released PD-L1 mRNAs were detected by qPCR. As for the detection of proteins, aptamer DNA probes were designed to recognize and anchor to membrane proteins (e.g., PD-L1, CD63) and then act as template molecules in the following qPCR assay for quantification. A lipid assay is also developed as an internal reference to normalize the amounts of samples in the qPCR of protein assay .…”