In cancer patients, chronic paclitaxel (PTX) treatment causes excruciating pain, limiting its use in cancer chemotherapy. The neuroprotective potential of synthetic cannabidiol (CBD) and CBD formulated in extracellular vesicles (CBD-EVs) isolated from human umbilical cord derived mesenchymal stem cells was investigated in C57BL/6J mice with PTX-induced neuropathic pain (PIPN). The particle size of EVs and CBD-EVs, surface roughness, nanomechanical properties, stability, and release studies were all investigated. To develop neuropathy in mice, PTX (8 mg/kg, i.p.) was administered every other day (four doses). In terms of decreasing mechanical and thermal hypersensitivity, CBD-EVs treatment was superior to EVs treatment or CBD treatment alone (p < 0.001). CBD and CBD-EVs significantly reduced mitochondrial dysfunction in dorsal root ganglions and spinal homogenates of PTX-treated animals by modulating the AMPK pathway (p < 0.001). Studies inhibiting the AMPK and 5HT1A receptors found that CBD did not influence the neurobehavioral or mitochondrial function of PIPN. Based on these results, we hypothesize that CBD and CBD-EVs mitigated PIPN by modulating AMPK and mitochondrial function.

Pulmonary lymphangioleiomyomatosis (LAM) is a metastatic sarcoma, but the mechanisms of LAM metastasis are unknown. Extracellular vesicles (EVs) regulate cancer metastasis; however, their roles in LAM have not yet been thoroughly investigated. Here, we report the discovery of distinct LAM-EV subtypes derived from primary tumor or metastasizing LAM cells that promote LAM metastasis through ITGα6/β1-c-Src-FAK signaling, triggered by the shuttling of ATP synthesis to cell pseudopodia or the activation of integrin adhesion complex, respectively. This signaling leads to increased LAM cell migration, invasiveness, and stemness and regulates metastable (hybrid) phenotypes that are all pivotal for metastasis. Mouse models corroborate in vitro data by demonstrating a significant increase in lung metastatic burden upon exposure to EVs through distinct mechanisms involving either lung resident fibroblasts or metalloproteinases' activation that are EV subtype dependent. The clinical relevance of these findings is underscored by increased EV biogenesis in LAM patients and the enrichment of these EV cargo with lung-tropic integrins and metalloproteinases. These findings establish EVs as a novel therapeutic target in LAM, warranting future clinical studies.

“Yan Li” was not included as an author in the original publication

The purpose of this study was to evaluate if phytocannabinoids, synthetic cannabidiol (CBD), and tetrahydrocannabivarin (THCV), and their combination, could protect mice from Paclitaxel-induced peripheral neuropathy (PIPN). Six groups of C57BL/6J mice (n = 6) were used in this study. The mice were given paclitaxel (PTX) (8 mg/kg/day, i.p.) on days 1, 3, 5, and 7 to induce neuropathy. Mice were evaluated for behavioral parameters, and dorsal root ganglions (DRG) were collected from the animals and subjected to RNA sequence and westernblot analysis at the end of the study. On cultured DRGs derived from adult male rats, immunocytochemistry and mitochondrial functional assays were also performed. When compared to individual treatments, the combination of CBD and THCV improved thermal and mechanical neurobehavioral symptoms in mice by twofold. Targets for CBD and THCV therapy were identified by KEGG (RNA sequencing). PTX reduced the expression of p-AMPK, SIRT1, NRF2, HO1, SOD2, and catalase while increasing the expression of PI3K, p-AKT, p-P38 MAP kinase, BAX, TGF-β, NLRP3 inflammasome, and caspase 3 in DRG homogenates of mice. Combination therapy outperformed monotherapy in reversing these protein expressions. To reduce mitochondrial superoxides, CBD and THCV were added to DRG primary cultures. WAY100135 and rimonabant altered the neuroprotective effects of CBD and THCV respectively by blocking 5-HT1A and CB1 receptors in mice and DRG primary cultures. The entourage effect of CBD and THCV against PIPN appears to protect neurons in mice via 5HT1A and CB1 receptors respectively.

Stem cell-derived EVs promote tissue repair [4], while cancer-derived EVs facilitate tumor growth and metastasis [5]. A landmark study by Lyden and colleagues (Nature Medicine, 2012) provided the first evidence that EVs derived from highly metastatic melanoma cells can permanently "educate" bone marrow progenitor cells, thereby enhancing the metastatic potential of primary tumors [6]. One of the most intriguing aspects of these vesicles is their ability to promote organotrophic metastasis, the phenomenon where EV cargo dictates the preferred organ destination of metastatic spread. Evidence suggests that this organotropism is largely governed by specific integrin subtypes carried on cancer-derived EVs. The Hoshino et al. (Nature, 2015) study elegantly demonstrated that EVs from tumor cells expressing α6β4 and α6β1 integrins are associated with lung metastasis, while those expressing αvβ5 integrins preferentially promote liver metastasis [7].Targeting these integrins significantly reduced EV uptake and the corresponding metastatic colonization. Mechanistically, α6β4 and α6β1 bind to laminin, activating c-SRC-S100A4 signaling and promoting lung-tropic metastasis, while αvβ5 binds to fibronectin on liver Kupffer cells, promoting liver-tropic metastasis. Additionally, αvβ3 and αvβ8 integrins in breast cancer EVs facilitate brain metastasis, whereas α4β1 and α5β1 integrins are implicated in bone metastasis [8,9]. In our recent studies, we found that ITGB1 expression in EVs derived from TSC2-null cells directed the formation of lung pre-metastatic niches in NCG mice. plasma-derived EVs from lymphangioleiomyomatosis (LAM) patients displayed elevated expression of ITGB1, c-SRC, FAK, and other integrin adhesion complex components, consistent with our in vitro findings [10].A central question remains: how do these nanosized vesicles so effectively reprogram recipient cells? Do they merely deliver molecular cargo, or do they carry pre-assembled protein complexes capable of instant signaling? Understanding EV topology, the spatial orientation of proteins within the vesicle membrane is crucial to answer this. EV integrins exist as heterodimers, and their cytoplasmic tails often bind to adaptor proteins such as talin and kindlin, which determine the activation state of the integrin receptor [11]. These integrin-adhesion complexes enable immediate communication with recipient cells, initiating rapid downstream signaling. Several studies have also demonstrated that cancer-derived EVs contain matrix metalloproteinases (MMP3, MMP7, MMP9, etc.), which enhance metastatic potential either by remodeling the extracellular matrix or by activating integrin signaling pathways [10,12,13]. Furthermore, EVs are enriched in tetraspanins (CD9, CD63, CD81)-classical EV markers whose expression correlates with EV biogenesis. Interestingly, tetraspanins themselves can contribute to the formation of focal adhesion points, amplifying the migratory and invasive potential of cancer cells [14,15].Two key determinants govern the oncogenic impact of EVs: how they are secreted and how they are internalized by recipient cells. Although the precise mechanisms of these processes are still being elucidated, several drugs are under investigation for their ability to inhibit EV biogenesis or uptake. Tipifarnib (a farnesyltransferase inhibitor) has been identified through high-throughput screening as a potent inhibitor of both ESCRT-dependent (Alix) and ESCRT-independent (nSMase2) EV biogenesis in metastatic castration-resistant prostate cancer cells [16]. GW4869, an inhibitor of neutral sphingomyelinase 2 (nSMase2), suppresses EV secretion and has been shown to inhibit proliferation and promote apoptosis in colorectal cancer models [17]. Cannabidiol (CBD) also reduces EV release and exhibits anticancer potential in various tumor types, though its specific mechanisms remain under investigation [18]. Dyngo-4A, a dynamin inhibitor, and its analog dynasore block EV uptake by disrupting clathrin-mediated endocytosis, thereby reducing tumor-derived EV internalization and angiogenesis [19,20]. While these findings highlight the promise of EV-targeted therapies, practical challenges remain mainly in difficulty of selectively targeting tumor-derived EVs without affecting physiological EVs. Regarding EV isolation, current methodologies include ultracentrifugation, ultrafiltration, size-exclusion chromatography, polymer-based precipitation, and affinity-based capture using antibodies have each with its advantages and limitations. According to the MISEV2023 guidelines (ISEV 2023), combining multiple methods is recommended to enhance purity, reproducibility, and yield [21]. Before functional assays, it is essential to verify cargo reproducibility and EV integrity across methods used to isolate EVs. Despite significant progress, key limitations persist, including the lack of standardized dose optimization and precise definitions of EV purity. The future of EV research depends on a deeper understanding of EV biogenesis, uptake mechanisms, and protein topology, as these factors determine vesicle function and specificity. In conclusion, effective targeting EVs in cancer requires a comprehensive understanding of their molecular cargo, topology, and signaling mechanisms. Therapeutic strategies that selectively modulate EV communication rather than indiscriminately blocking biogenesis or uptake are more likely to yield safe and successful outcomes. Looking forward, microfluidic chip technologies hold promises for rapid, noninvasive tumor detection from a single drop of patient blood, marking an exciting frontier in cancer diagnostics and precision medicine using EVs.

Currently, increasing research on the novel drug delivery system (DDS) is emerging, especially in the stimuli-based nano carriers. Among other types of external and internal stimuli, Redox-responsive delivery systems are a novel and promising approach for targeted therapy that takes advantage of the distinct oxidative stress and redox gradients present in target disease microenvironments. These systems are designed to exploit the higher levels of reactive oxygen species (ROS) and reducing agents such as glutathione (GSH) found in target disease cells compared to normal cells. These cutting-edge redox-responsive DDS based on nanocarriers have enhanced treatment efficacy. Redox-responsive nanoshells are a type of stimuli-responsive nanomaterial that undergo structural or functional changes in response to the redox environment. These nanoshells are particularly useful in drug delivery, cancer therapy, and biosensing due to their ability to release cargo selectively in reducing environments, such as the intracellular environment of cancer cells. By improving their design and getting over current obstacles, they could revolutionize target treatment by providing a more individualized, safe, and efficient therapeutic approach. The chapter discusses redox-responsive nanoshells, application, and next-generation advancements. It also covers design approaches for redox-responsive nanoshells, from conventional to advanced redox-responsive nanoshells for targeted DDS. It could be beneficial to the new researchers working in the field of advanced delivery research.