Published online Jun 15, 2026. doi: 10.4251/wjgo.v18.i6.117697
Revised: January 21, 2026
Accepted: February 25, 2026
Published online: June 15, 2026
Processing time: 175 Days and 17.7 Hours
Gastric cancer remains a major cause of cancer-related mortality worldwide, and traditional chemotherapy is frequently limited by poor tumor targeting, off-target toxicity, and insufficient combined therapeutic benefit. Emerging evidence suggests that tumor microenvironmental cues, particularly the acidic condition, can enhance the tumor-targeting capacity and modulate the bioactive cargo of extracellular vesicles (EVs) derived from certain cell types. However, whether low-pH preconditioned macrophage-derived EVs (LP-EV) acquire enhanced tumor-targeting properties and intrinsic anti-tumor activity, as well as the under
To investigate tumor targeting and intrinsic bioactivity of LP-EV, and their com
LP-EV were isolated from J774.1 macrophages cultured at pH 6.5 and systematically characterized in comparison with control EVs (C-EV). Tumor-targeting performance was evaluated using cellular uptake assays and in vivo fluorescence imaging. miRNA profiles of LP-EV and C-EV were analyzed by sequencing. PTX-loaded LP-EV (PTX@LP-EV) were prepared via sonication, and their anti-tumor effects were assessed in vitro and in MKN45 gastric cancer xenograft models. Tumor transcriptome sequencing was conducted to elucidate mechanisms associated with the enhanced anti-tumor effects.
LP-EV exhibited significantly enhanced tumor-targeting ability compared with C-EV. miRNA sequencing revealed that LP-EV were selectively enriched in miRNAs with previously reported anti-tumor activities. PTX@LP-EV exhibited markedly superior anti-tumor efficacy compared with free PTX, accompanied by pronounced tumor growth inhibition. Transcriptome analysis revealed that PTX@LP-EV induced a distinct transcriptional profile compared to free PTX, and gene set enrichment analysis indicated the engagement of cooperative biological pathways consistent with enhanced therapeutic effects. A miRNA-target gene regulatory network analysis suggested that LP-EV-derived miRNAs may contribute to the remodeling of tumor survival and stress-response networks, thereby suggesting a potential molecular basis for the observed combination therapeutic effects.
Low-pH preconditioning confers macrophage-derived EVs with improved tumor-targeting capability and intrinsic anti-tumor activity. PTX@LP-EV achieve enhanced anti-tumor efficacy through coordinated, multi-level transcriptional reprogramming in gastric cancer.
Core Tip: In this study, we report a microenvironment-inspired strategy to customize macrophage-derived extracellular vesicles (EVs) through low-pH preconditioning, mimicking the acidic characteristics of the tumor microenvironment. This approach markedly enhances EVs' tumor targeting and endows them with a distinct miRNA cargo associated with reported anti-tumor functions. When used as nanocarriers for paclitaxel (PTX), the resulting biomimetic system (PTX@LP-EV) achieves superior anti-tumor efficacy in gastric cancer models. Mechanistically, EV-borne miRNAs induced by tumor microenvironmental stimulation may cooperatively suppress pro-tumorigenic genes, which is consistent with the observed enhanced therapeutic effects.
- Citation: Wang JH, Hu ML, Shi M, Ma JL, Yang J, Wang YG. Low-pH preconditioned macrophage-derived extracellular vesicles enable targeted and enhanced paclitaxel therapy in gastric cancer. World J Gastrointest Oncol 2026; 18(6): 117697
- URL: https://www.wjgnet.com/1948-5204/full/v18/i6/117697.htm
- DOI: https://dx.doi.org/10.4251/wjgo.v18.i6.117697
Gastric cancer remains one of the leading causes of cancer-related mortality worldwide, particularly in East Asia[1,2]. Although chemotherapy represents a cornerstone in the treatment of advanced or metastatic gastric cancer, chemotherapeutic agents, such as paclitaxel (PTX), are frequently limited by non-specific biodistribution and systemic toxicity, which restrict effective drug accumulation within tumors and compromise therapeutic efficacy[3,4]. In addition, the inability of conventional chemotherapy to elicit coordinated anti-tumor mechanisms further limits overall treatment outcomes[5,6], underscoring the urgent need for more precise and efficient drug delivery strategies to improve therapeutic outcomes.
Extracellular vesicles (EVs) are naturally occurring nanoscale lipid bilayer vesicles secreted by almost all cell types and play a critical role in intercellular communication by transporting bioactive cargos including proteins, lipids, and nucleic acids[7-9]. Owing to their intrinsic biocompatibility, low immunogenicity, excellent tissue penetration, and ability to traverse biological barriers, EVs have emerged as a highly promising drug delivery platform for cancer therapy[10-12]. Among the diverse cellular sources of EVs, macrophage-derived EVs have attracted increasing attention. Macrophages engage in dynamic crosstalk with tumor cells through cytokines, chemokines[12], and exhibit a pronounced chemotactic capacity to infiltrate tumor tissues in response to signals from the tumor microenvironment[13-15]. Importantly, this tumor-homing property is believed to be inherited by the EVs they secrete, enabling macrophage-derived EVs to preferentially accumulate within tumor tissues[16,17]. Moreover, the lipid bilayer structure of EVs facilitates the encapsulation of hydrophobic chemotherapeutic drugs such as PTX, providing a biologically favorable platform to overcome solubility limitations, rapid clearance, and off-target toxicity associated with conventional formulations[12,18,19].
Despite these advantages, the tumor-targeting efficiency and overall therapeutic performance of native, unmodified EVs remain suboptimal. In recent years, functional customization of EVs through in vitro preconditioning of parent cells has emerged as an effective strategy to enhance EV bioactivity and targeting capability[20]. Notably, previous studies have demonstrated that simulating key features of the tumor microenvironment can markedly improve the tumor-homing ability of tumor cell-derived EVs[21]. Among various microenvironmental stimuli, acidic preconditioning has been shown to be particularly effective. EVs derived from tumor cells exposed to acidic condition exhibit superior tumor-targeting capacity compared with those under other stimuli. Beyond enhancing physical targeting, acidic preconditioning profoundly influences cellular phenotype and reshapes EV biogenesis as well as cargo composition[22,23]. In this context, the identity of the parental cell critically determines how microenvironmental cues are sensed and translated into EV functional outputs[24]. Macrophages are professional sensors of extracellular signals and exhibit remarkable phenotypic and molecular plasticity in response to microenvironmental stressors, including acidic pH[24,25]. Such intrinsic ad
Based on this rationale, this study aimed to investigate the biological characteristics and therapeutic potential of low-pH preconditioned macrophage-derived EVs (LP-EV) in gastric cancer. We systematically characterized LP-EV and compared them with control EVs (C-EV) produced under physiological pH condition, with a particular focus on tumor-targeting capability. Through small RNA sequencing, we analyzed the miRNA expression profiles of these EVs and explored the potential functional alterations of LP-EV. Furthermore, we encapsulated PTX into LP-EV to construct a biomimetic nanodelivery system (PTX@LP-EV) and comprehensively evaluated its anti-tumor efficacy and underlying cooperative mechanisms. This work provides a tumor microenvironment-inspired, preconditioned EV-based strategy for targeted combination therapy and offers new insights into the rational design of precision nanomedicine for gastric cancer treatment.
The murine macrophage cell line J774.1 and the human gastric carcinoma cell line MKN45 were obtained from the Cell Resource Center of Peking Union Medical College. J774.1 macrophages were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; Gibco). For acidic preconditioning, cells were cultured in complete medium adjusted to pH 6.5 for 24 hours to generate LP-EV. C-EV were collected from cells maintained under physiological pH conditions (pH 7.4). All cell cultures were incubated at 37 °C in a humidified atmosphere containing 5% CO2.
Conditioned culture media were harvested and cleared of cells and debris by sequential centrifugation at 300 × g for 10 minutes, 2000 × g for 30 minutes, and 10000 × g for 45 minutes. The resulting supernatant was passed through a 0.22-μm membrane filter and subsequently subjected to ultracentrifugation at 100000 × g for 70 minutes at 4 °C. The EV pellet was resuspended in PBS and washed by a second round of ultracentrifugation under identical conditions. Purified EVs were finally resuspended in PBS and stored at -80 °C until further use.
EVs morphology was examined using transmission electron microscopy (TEM). The expression of EV-associated markers (ALIX, CD9, and CD63) was assessed by western blotting using antibodies purchased from Abcam. Particle size distribution and surface zeta potential were analyzed by nanoparticle tracking analysis (NTA).
Purified C-EV and LP-EV were fluorescently labeled with the lipophilic dye DiI (Beyotime). These EVs were incubated with MKN45 cells for 24 hours. Then the cells were thoroughly washed with PBS and fixed with 4% paraformaldehyde. Cytoskeletal F-actin was visualized using fluorescent phalloidin, and nuclei were counterstained with DAPI. Fluorescence signals were captured via confocal laser scanning microscope.
Female BALB/c nude mice (4-6 weeks old, 15-20 g) were purchased from Vital River Laboratories. All mice were maintained under specific pathogen-free (SPF) conditions in individually ventilated cages (5 mice per cage). The housing environment was controlled with a 12-hour light/dark cycle, temperature of 22 ± 2 °C, and relative humidity of 50%-60%. Standard chow and water were provided ad libitum. To establish the subcutaneous xenograft model, MKN45 cells (2 × 106 cells suspended in 100 μL PBS) were injected subcutaneously into the left flank of each mouse. Tumor growth was monitored every three days using digital calipers.
When the subcutaneous tumors reached approximately 100 mm3, tumor-bearing mice were randomly assigned to experimental groups. Mice received a single intravenous injection of DiR-labeled C-EV or LP-EV (100 μg per mouse). Whole-body fluorescence imaging was performed at 2, 4, 8, 12, 24, 48, and 72 hours post-injection using an IVIS Spectrum imaging system (PerkinElmer). At 72 hours, mice were euthanized, and major organs (heart, liver, spleen, lungs, kidneys) as well as tumor tissues were excised for ex vivo fluorescence imaging. Tumor-associated fluorescence intensity was quantified using Living Image software.
Total RNA was isolated from C-EV and LP-EV using TRIzol reagent (Invitrogen). Construction of small RNA libraries and high-throughput sequencing were performed on an Illumina NovaSeq 6000 platform by a commercial service provider. Differential miRNA expression analysis was carried out using the DESeq2 package, with thresholds set at |log2 fold change| > 1 and adjusted P < 0.05. Putative target genes of differentially expressed miRNAs were predicted using both miRanda and TargetScan databases. These analyses were performed in an exploratory manner to identify differentially enriched miRNAs and potential functional trends associated with low-pH preconditioning. Functional enrichment analyses were conducted using the clusterProfiler R package.
Total RNA was isolated from C-EV and LP-EV using a miRNA-specific extraction kit according to the manufacturer’s instructions. The concentration and purity of RNA were assessed using a NanoDrop spectrophotometer. Reverse transcription of miRNAs was performed using a miRNA reverse transcription kit with stem-loop primers. Quantitative real-time PCR (qPCR) was conducted using SYBR Green-based chemistry on a real-time PCR system. The expression levels of miR-204-3p, miR-708-3p, and miR-324-5p were quantified. U6 small nuclear RNA was used as an internal control for normalization. Relative miRNA expression levels were calculated using the 2−ΔΔCt method. All reactions were performed in triplicate, and results were presented as mean ± SD.
To evaluate the functional relevance of EV-enriched miRNAs, MKN45 gastric cancer cells were seeded into six-well plates and allowed to reach approximately 60%-70% confluence. Cells were then transfected with a synthetic miR-204-3p mimic or a corresponding negative control mimic using a lipid-based transfection reagent, following the manufacturer’s protocol. After 24 hours of transfection, cells were treated with C-EV in combination with PTX for an additional 24 hours. The concentration of PTX and EVs used in this assay was consistent with that applied in the main in vitro anti-tumor experiments.
Apoptosis was subsequently assessed by flow cytometry using an Annexin V-FITC/propidium iodide (PI) apoptosis detection kit. Cells were collected, washed with cold PBS, stained according to the manufacturer’s instructions, and analyzed using a flow cytometer. The proportions of early and late apoptotic cells were quantified and compared between groups.
PTX was loaded into LP-EV using a sonication-assisted method. Briefly, LP-EV were mixed with PTX solution and subjected to ultrasonic treatment using an ultrasonic cell disruptor (Scientz-IID) in an ice bath at 200 W for 5 minutes with intermittent pulses (15 seconds on/15 seconds off). Unencapsulated PTX was removed by ultrafiltration, yielding PTX-loaded LP-EV (PTX@LP-EV).
Drug loading capacity and encapsulation efficiency (EE) were determined by high-performance liquid chromatography. Chromatographic conditions: C18 column, mobile phase acetonitrile:water (50:50, v/v), flow rate 1.0 mL/minute, detection wavelength 227 nm. Calculations were performed using the standard curve method.
The EE and loading content (LC) were calculated based on actual drug content (ADC) according to the following equations:
EE (%) = (ADC/PTX added) × 100%.
LC (%) = (ADC/LP-EV) × 100%.
Cell viability was evaluated using the Cell Counting Kit-8 (CCK-8) assay. MKN45 cells were plated in 96-well plates and exposed to PBS, LP-EV, free PTX, or PTX@LP-EV for 48 hours. Subsequently, CCK-8 solution was added to each well and incubated for an additional 2 hours. Absorbance was recorded at 450 nm using a microplate reader, and relative cell viability was calculated by normalizing to the untreated control after subtraction of background values.
Apoptotic cell death was assessed by flow cytometry using an Annexin V-FITC/PI apoptosis detection kit (BD Biosciences), with two independent experimental settings corresponding to different study objectives. To evaluate whether LP-EV could enhance PTX-induced apoptosis, MKN45 cells were treated with PBS, PTX combined with C-EV (PTX + C-EV), or PTX combined with LP-EV (PTX + LP-EV), followed by Annexin V-FITC/PI staining and flow cyto
To evaluate the systemic safety of EV-based and PTX-based formulations, healthy nude mice without tumor implantation were randomly divided into corresponding treatment groups. Mice received intravenous injections of PBS or LP-EV, following the same dosing schedule used in the anti-tumor efficacy study. At the completion of treatment, blood samples were collected for hematological analysis and serum biochemistry to assess liver function (alanine aminotransferase and aspartate aminotransferase) and renal function (creatinine and urea). Major organs were harvested, fixed, embedded in paraffin, sectioned, and subjected to hematoxylin and eosin (H&E) staining to examine potential treatment-related histopathological alterations. No unexpected adverse events or treatment-related mortality were observed during the study period.
When the subcutaneous tumors reached approximately 100 mm3, tumor-bearing mice were randomly assigned to four treatment groups (n = 6 per group): PBS, LP-EV, free PTX, and PTX@LP-EV. Treatments were administered via intravenous injection through the tail vein every three days for a total of six doses. Tumor growth was monitored throughout the treatment period by measuring tumor length and width using a digital caliper at three-day intervals. Tumor volume was calculated according to the following formula: V = 0.5 × length × width2. Investigators responsible for tumor measurement and data analysis were blinded to group allocation throughout the study. All animal experiments were performed during the light phase of the light/dark cycle. Mice were euthanized on day 25, and tumor tissues were excised, and sectioned for histopathological analysis. Tumor cell proliferation and apoptosis were evaluated by immunohistochemical staining for Ki-67 and caspase-3, respectively, while apoptotic cell death was further examined using TUNEL staining.
Tumor tissues from PBS, free PTX, and PTX@LP-EV groups (n = 2 per group) were collected for total RNA extraction and transcriptome sequencing. Sequencing reads were aligned to the reference genome using HISAT2, and gene expression levels were quantified with featureCounts. Differentially expressed genes were identified using DESeq2 with |log2 fold change| > 1 and adjusted P < 0.05. Gene set enrichment analysis (GSEA) was conducted using clusterProfiler to explore global transcriptional programs associated with different treatments. These transcriptomic analyses were designed as exploratory analyses to identify treatment-associated transcriptional patterns and generate mechanistic hypotheses, rather than to establish definitive causal relationships. Predicted targets of upregulated miRNAs in LP-EV were in
Statistical analyses were conducted with GraphPad Prism 8.4.3. Statistical analysis was performed using unpaired t test for comparing two normally distributed groups, one-way ANOVA for comparing more than two normally distributed groups. Significant values were indicated as aP < 0.05, bP < 0.01, cP < 0.001, and dP < 0.0001.
Based on the acidic characteristics of the tumor microenvironment and commonly adopted in vitro acidic stimulation models reported in previous studies, pH 6.5 was selected to mimic tumor microenvironment acidity[26-28]. Using this acidic condition allowed us to effectively mimic the stress encountered in highly acidic regions of tumors and to examine how extracellular acidity influences the biological properties of macrophage-derived EVs. Accordingly, J774.1 macro
TEM revealed that LP-EV exhibited the typical cup-shaped morphology of EVs, with intact limiting membranes and no obvious aggregation or structural disruption (Figure 1B). Western blot analysis confirmed high expression of characteristic EV marker proteins ALIX, CD9, and CD63 in both C-EV and LP-EV (Figure 1C). NTA indicated that the particle sizes of LP-EV and C-EV were mainly distributed between 50 nm and 200 nm, with an average diameter of approximately 135 nm, consistent with canonical EV size ranges (Figure 1D and Supplementary Figure 1). To evaluate the storage stability of LP-EV, particles were resuspended in either plain PBS or PBS supplemented with 10% FBS and kept at 4 °C for up to 7 days. NTA measurements taken at different time points indicated no significant alteration in particle size under both conditions, demonstrating that LP-EV remain physically stable during short-term cold storage (Figure 1E).
To evaluate the tumor-targeting ability of LP-EV, we first conducted in vitro cellular uptake experiments. C-EV and LP-EV were labeled with the fluorescent dye DiI and co-incubated with gastric cancer cells MKN45. Confocal microscopy observation showed that compared to the C-EV group, LP-EV were internalized by MKN45 cells with significantly higher efficiency, as indicated by stronger intracellular red fluorescence signals (Figure 1F). Subsequently, we evaluated the in vivo targeting ability in an MKN45 subcutaneous xenograft model in nude mice. After intravenous injection of DiR-labeled C-EV or LP-EV, small animal live imaging at each time point post-injection (2, 4, 8, 12, 24, 48, and 72 hours) showed that both types of EVs achieved accumulation at the tumor site. However, the fluorescence intensity in the tumor region of the LP-EV group was significantly higher than that of the C-EV group (Figure 1G and H). Mice were sacrificed at 72 hours, and major organs as well as tumor tissues were harvested for ex vivo imaging. Results showed that the accumulation of LP-EV in tumor tissues was significantly greater than that of C-EV (Figure 1I). Collectively, these data indicate that low-pH preconditioning markedly improves both cellular uptake and in vivo tumor accumulation of macrophage-derived EVs. The preferential tumor accumulation of LP-EV suggests that low-pH preconditioning may modulate EV-tumor interactions, contributing to enhanced tumor retention.
Beyond enhanced tumor targeting, EVs may also exert biological effects through their internal molecular cargo, among which miRNAs are key functional mediators. To determine whether low-pH preconditioning reshapes the miRNA composition of macrophage-derived EVs, small RNA sequencing was performed on C-EV and LP-EV. Principal component analysis (PCA) showed clear separation between the miRNA expression profiles of C-EV and LP-EV, indicating significant differences in their miRNA composition (Figure 2A). Differential analysis identified multiple miRNAs upregulated in LP-EV (Figure 2B and Supplementary Figure 2). Notably, several of these upregulated miRNAs have been previously reported to possess anti-tumor activities. For example, miR-708-3p inhibits epithelial-mesenchymal transition, reducing metastasis and chemoresistance in breast cancer[29]; miR-204-3p inhibits cell proliferation and promotes apoptosis in gastric cancer, involving the inhibition of MAPK and necroptosis pathways[30]; miR-324-5p acts as a tumor suppressor in gastric cancer, reducing cell viability and inducing apoptosis by targeting TSPAN8[31]. Further prediction of target genes and Gene Ontology functional enrichment analysis revealed that the upregulated miRNAs in LP-EV are predicted to target genes involved in key biological processes. Notably, the significant enrichment of processes such as “negative regulation of apoptotic process”, “DNA repair”, “angiogenesis”, and “regulation of cell cycle” suggests that these miRNAs may collectively inhibit pro-survival, proliferative, and metabolic pathways in tumor cells, thereby potentially exerting coordinated anti-tumor effects (Figure 2C). Collectively, these results suggest that low-pH preconditioning not only enhances the tumor-homing capability of macrophage-derived EVs but also confers them with a distinct miRNA cargo that may contribute to coordinated anti-tumor effects.
Based on these observations, we further evaluated whether LP-EV could enhance the effect of PTX in vitro. Flow cytometric analysis demonstrated that the PTX + LP-EV combination induced significantly higher apoptosis rates in MKN45 cells compared with PTX combined with C-EV (Figure 2D), indicating that LP-EV contribute biologically active components capable of potentiating chemotherapeutic efficacy.
To verify the reliability of the sequencing data, qPCR was performed to validate representative miRNAs that were significantly enriched in LP-EV. Consistent with the sequencing results, qPCR analysis confirmed that miR-708-3p, miR-204-3p, and miR-324-5p were all significantly upregulated in LP-EV compared with C-EV (Figure 2E). Among these candidates, miR-204-3p exhibited the highest relative increase in LP-EV. To further explore the potential functional relevance of miR-204-3p, a miRNA mimic-based assay was conducted. MKN45 cells were transfected with a synthetic miR-204-3p mimic or a negative control mimic, followed by treatment with C-EV combined with PTX. Flow cytometric analysis demonstrated that enforced expression of miR-204-3p significantly enhanced PTX-induced apoptosis compared with the control group (Figure 2F and Supplementary Figure 3). These results indicate that miR-204-3p, one of the miRNAs enriched in LP-EV, is functionally capable of modulating tumor cell sensitivity to chemotherapy.
Building on the enhanced tumor-targeting capability and the distinct miRNA profile of LP-EV, we further employed LP-EV as a nanocarrier for PTX delivery. PTX-loaded LP-EV (PTX@LP-EV) were prepared via sonication method (Figure 3A). TEM observation showed that PTX@LP-EV maintained intact vesicle structure after PTX loading (Figure 3B). High-performance liquid chromatography analysis confirmed the presence of a characteristic peak at the retention time of the PTX standard in the lysate of PTX@LP-EV, indicating successful PTX loading (Figure 3C), with a LC of 15.95% (Supplementary Table 1). NTA analysis showed that the average particle size and the zeta potential of PTX@LP-EV did not change significantly, compared to empty LP-EV (Figure 3D).
The in vitro anti-tumor activity of PTX@LP-EV was subsequently evaluated in MKN45 cells. Flow cytometric analysis of apoptosis showed that PTX@LP-EV treatment induced a significantly higher proportion of apoptotic cells than free PTX or LP-EV alone at equivalent PTX concentrations (Figure 3E). Consistently, CCK-8 assays revealed that PTX@LP-EV exerted a more pronounced inhibitory effect on tumor cell proliferation compared with free PTX or LP-EV treatment (Figure 3F). Together, these findings indicate that PTX@LP-EV induces an enhanced anti-tumor effect in vitro, leading to increased apoptosis and inhibition of tumor cell proliferation.
Prior to in vivo application of PTX@LP-EV, the biosafety of its carrier, LP-EV, was first evaluated. Healthy nude mice were intravenously administered LP-EV at the same dose used in the treatment group. Histological examination of major organs, including the heart, liver, spleen, lungs, and kidneys, revealed no apparent pathological abnormalities based on H&E staining (Figure 4A). In addition, hematological analysis demonstrated that complete blood count parameters, as well as liver- and kidney function-related biochemical indices, remained within normal ranges and showed no significant differences compared with the PBS control group (Figure 4B and C). Collectively, these results indicate that LP-EV exhibits favorable biocompatibility and an acceptable safety profile in vivo.
Subsequently, we evaluated the therapeutic effect of PTX@LP-EV in an MKN45 tumor-bearing nude mouse model (Figure 5A). Tumor-bearing mice were randomly divided into four groups and treated with PBS, LP-EV alone, free PTX (administered at the same PTX dose as that encapsulated in PTX@LP-EV), or PTX@LP-EV, respectively. The tumor growth curve showed that compared to the PBS group, the LP-EV group had limited inhibitory effect on tumor growth, whereas treatment with free PTX resulted in moderate tumor growth suppression. In contrast, tumor growth in the PTX@LP-EV group was most markedly inhibited (Figure 5B). Consistently, the average tumor weight at the end of the observation period was significantly lower in the PTX@LP-EV group than in the other treatment groups (Figure 5C). Immunohistochemical analysis of tumor tissues revealed that the PTX@LP-EV group had the lowest expression level of the proliferation marker Ki-67, while the proportions of cells positive for the apoptosis marker cleaved caspase-3 and TUNEL were the highest (Figure 5D). The above results demonstrate that PTX@LP-EV effectively inhibited tumor growth in vivo, with efficacy superior to that of the free drug or carrier alone.
To further explore the molecular mechanisms associated with the enhanced anti-tumor activity of PTX@LP-EV, tumor tissues from treated mice were subjected to transcriptome sequencing analysis. PCA analysis showed clear clustering of samples from the PBS, free PTX, and PTX@LP-EV groups, with significant inter-group differences (Figure 6A). Venn diagram analysis further revealed that PTX@LP-EV treatment triggered a large number of unique gene expression that did not occur with free PTX or LP-EV treatment alone (Figure 6B). GSEA revealed distinct transcriptional programs induced by the two treatments. Compared to the PBS group, free PTX predominantly enriched apoptosis-related pathways (Figure 6C). Meanwhile, the pathway negative regulation of extrinsic apoptotic signaling pathway via death domain receptors was negatively enriched (Supplementary Figure 4), further supporting the involvement of apoptotic processes in PTX-treated tumors. In contrast, PTX@LP-EV treatment triggered a more complex and unique response compared to free PTX alone (Figure 6D). This included the significant upregulation of pathways involved in intracellular vesicle transport (e.g., cytoplasmic vesicle lumen), type I interferon-mediated signaling and response, and DNA damage repair (e.g., homologous recombination). Concurrently, PTX@LP-EV led to the marked downregulation of pathways supporting tumor cell resilience, such as glutathione metabolism, lipid and fatty acid oxidation, as well as multiple ribosome- and translation-related processes. Differential gene heatmap analysis further identified a set of genes specifically downregulated in the PTX@LP-EV group, but not in the free PTX group (Figure 6E). Key examples include Dgki (overexpressed in colon and gastric cancers and linked to poor prognosis[32-34]) and Gclm (implicated in platinum-based chemotherapy resistance in colorectal cancer cells[35]). The specific suppression of these pro-tumorigenic genes underscores the distinct and combined anti-tumor effect of the LP-EV delivery system beyond the action of free PTX. Interestingly, we integrated the predicted target genes of upregulated miRNAs in LP-EV with the downregulated differentially expressed genes identified in the PTX@LP-EV group, and visualized the resulting miRNA-target gene regulatory network using a Sankey diagram. The analysis revealed that multiple miRNAs enriched in LP-EV, including miR-708-3p, miR-204-3p, and miR-324-5p, potentially targeted a range of key genes that were suppressed in the PTX@LP-EV-treated tumors (Figure 6F). These findings suggest that functional miRNAs delivered by LP-EV may cooperate with PTX by repressing genes associated with tumor cell survival and drug resistance, thereby enhancing overall anti-tumor efficacy.
In this study, we developed and validated a biomimetic chemotherapy delivery platform based on macrophage-derived EVs conditioned under a simulated acidic tumor microenvironment. Our findings indicated that low-pH preconditioning conferred macrophage-derived EVs with improved tumor-homing capacity, a distinct miRNA cargo profile, and intrinsic biological activity that enhanced the therapeutic effect of PTX, culminating in superior anti-tumor efficacy of the integrated PTX@LP-EV system.
Efficient tumor targeting remains a central bottleneck in EV-based drug delivery. Beyond conventional engineering strategies that introduce exogenous targeting ligands onto EV surfaces, preconditioning parent cells to reflect cues of the tumor microenvironment has emerged as an effective approach to enhance the tumor tropism of EVs. Among these cues, acidic condition has been reported to exert the most pronounced effects on the targeting behavior of tumor cell-derived EVs[21]. While acidic preconditioning has been reported to enhance the tumor tropism of tumor cell-derived EVs, our data demonstrate that acidic condition similarly augments the tumor-homing behavior of macrophage-derived EVs, underscoring the generalizability of acidity-driven EV functional customization across different cellular origins. Future proteomic profiling of LP-EV surface components may further elucidate the molecular determinants underlying this enhanced targeting performance.
In addition to altering targeting behavior, acidic condition has also been shown to reshape the bioactive cargo of EVs, which may confer potential therapeutic potential[22,36-38]. Consistent with this paradigm, miRNA profiling revealed that LP-EV are enriched in multiple miRNAs with reported tumor-suppressive functions, such as apoptosis, stress adaptation, and metabolic regulation. Several of these miRNAs-including miR-204-3p and miR-324-5p-have been reported to inhibit MAPK signaling or TSPAN8-mediated oncogenic pathways[30,31], respectively. Notably, these miRNAs were also predicted to target genes specifically downregulated in PTX@LP-EV treated tumors, such as Dgki, a gene overexpressed in gastric and colorectal cancers and associated with poor prognosis[32-34]. Collectively, these observations suggest that macrophage-derived EVs exhibit a particularly pronounced functional response to acidic condition, potentially attributable to the intrinsic environmental responsiveness of macrophages. And these findings provide mechanistic support for the concept that LP-EV function as biologically active co-therapeutic agents, rather than inert drug carriers.
From a drug delivery perspective, EVs are particularly well suited for hydrophobic chemotherapeutic agents such as PTX. Conventional PTX formulations depend on solubilizing excipients that contribute to systemic toxicity and limit therapeutic windows[39-41]. In contrast, the lipid bilayer architecture of EVs enables efficient encapsulation, preferential tumor-site delivery, and reduced off-target exposure[42-44]. By integrating acidic preconditioning with EV-mediated PTX loading, the PTX@LP-EV platform achieved enhanced tumor accumulation and realized an integrated therapeutic benefit at the system level.
Collectively, our findings support a therapeutic platform in which LP-EV enhance chemotherapy efficacy through multi-layered integration, encompassing improved tumor targeting, biologically active cargo remodeling, and optimized drug delivery. This work highlights the potential of tumor microenvironment-inspired EV engineering as a versatile and clinically translatable strategy for precision oncology. Future studies focusing on cargo-function validation and therapeutic scalability will further advance the translational prospects of this platform.
In summary, this study establishes a microenvironment-inspired strategy to functionally customize macrophage-derived EVs by acidic preconditioning, mimicking key features of the tumor microenvironment. The resulting LP-EV exhibit markedly enhanced tumor-targeting capability and a distinct miRNA cargo profile associated with reported anti-tumor activity. When employed as nanocarriers for PTX delivery, LP-EV enable efficient tumor accumulation and achieve superior anti-tumor efficacy through coordinated, multi-level mechanisms in a gastric cancer model. Collectively, this work not only provides a biomimetic nanoplatform to address key challenges in chemotherapeutic drug delivery, but also advances the understanding of how microenvironmental cues regulate EV function, offering a rational foundation for the development of precision and personalized EV-based cancer therapies.
| 1. | Sundar R, Nakayama I, Markar SR, Shitara K, van Laarhoven HWM, Janjigian YY, Smyth EC. Gastric cancer. Lancet. 2025;405:2087-2102. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 187] [Cited by in RCA: 191] [Article Influence: 191.0] [Reference Citation Analysis (0)] |
| 2. | Bray F, Laversanne M, Sung H, Ferlay J, Siegel RL, Soerjomataram I, Jemal A. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024;74:229-263. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 16785] [Cited by in RCA: 15073] [Article Influence: 7536.5] [Reference Citation Analysis (23)] |
| 3. | Lordick F, Shitara K, Janjigian YY. New agents on the horizon in gastric cancer. Ann Oncol. 2017;28:1767-1775. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 104] [Cited by in RCA: 95] [Article Influence: 10.6] [Reference Citation Analysis (0)] |
| 4. | Pao YS, Liao KJ, Shiau YC, Chao MH, Li MC, Lin LM, Chang HH, Yeh HW, Chen YJ, Chiu YT, Pan MY, Chang YH, Shen SY, Lin SY, Cheng HC, Lin YC, Sun YJ, Kuo CC, Hsieh HP, Wang LH. KIF2C promotes paclitaxel resistance by depolymerizing polyglutamylated microtubules. Dev Cell. 2025;60:2097-2113.e8. [RCA] [PubMed] [DOI] [Full Text] [Cited by in RCA: 7] [Reference Citation Analysis (0)] |
| 5. | Wang M, Qiu R, Yu S, Xu X, Li G, Gu R, Tan C, Zhu W, Shen B. Paclitaxel-resistant gastric cancer MGC-803 cells promote epithelial-to-mesenchymal transition and chemoresistance in paclitaxel-sensitive cells via exosomal delivery of miR-155-5p. Int J Oncol. 2019;54:326-338. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 28] [Cited by in RCA: 74] [Article Influence: 9.3] [Reference Citation Analysis (0)] |
| 6. | Hsieh CH, Huang YW, Tsai TF. Oral Conventional Synthetic Disease-Modifying Antirheumatic Drugs with Antineoplastic Potential: a Review. Dermatol Ther (Heidelb). 2022;12:835-860. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in RCA: 5] [Reference Citation Analysis (0)] |
| 7. | van Niel G, D'Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nat Rev Mol Cell Biol. 2018;19:213-228. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 7245] [Cited by in RCA: 6344] [Article Influence: 793.0] [Reference Citation Analysis (7)] |
| 8. | Kalluri R, LeBleu VS. The biology, function, and biomedical applications of exosomes. Science. 2020;367:eaau6977. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 9106] [Cited by in RCA: 8001] [Article Influence: 1333.5] [Reference Citation Analysis (13)] |
| 9. | Wang J, Jia R, Wei W, Hu M, Li F, Wang W, Ye P, Zhao J, Xu L, Wang S, Wang Y, Shi M, Ma G. Spleen-liver dual accumulation of ly6clowExo potentiates synergistic immune modulation for liver fibrosis therapy. Nano Today. 2024;58:102422. [RCA] [DOI] [Full Text] [Cited by in RCA: 1] [Reference Citation Analysis (0)] |
| 10. | Herrmann IK, Wood MJA, Fuhrmann G. Extracellular vesicles as a next-generation drug delivery platform. Nat Nanotechnol. 2021;16:748-759. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 1665] [Cited by in RCA: 1380] [Article Influence: 276.0] [Reference Citation Analysis (3)] |
| 11. | Ahmadi M, Rezaie J. Tumor cells derived-exosomes as angiogenenic agents: possible therapeutic implications. J Transl Med. 2020;18:249. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 126] [Cited by in RCA: 117] [Article Influence: 19.5] [Reference Citation Analysis (0)] |
| 12. | Yang C, Xue Y, Duan Y, Mao C, Wan M. Extracellular vesicles and their engineering strategies, delivery systems, and biomedical applications. J Control Release. 2024;365:1089-1123. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 157] [Cited by in RCA: 121] [Article Influence: 60.5] [Reference Citation Analysis (0)] |
| 13. | Liu Y, Zhang L, Ju X, Wang S, Qie J. Single-Cell Transcriptomic Analysis Reveals Macrophage-Tumor Crosstalk in Hepatocellular Carcinoma. Front Immunol. 2022;13:955390. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 1] [Cited by in RCA: 25] [Article Influence: 6.3] [Reference Citation Analysis (0)] |
| 14. | Pang L, Zhu Y, Qin J, Zhao W, Wang J. Primary M1 macrophages as multifunctional carrier combined with PLGA nanoparticle delivering anticancer drug for efficient glioma therapy. Drug Deliv. 2018;25:1922-1931. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 75] [Cited by in RCA: 102] [Article Influence: 12.8] [Reference Citation Analysis (0)] |
| 15. | Crone SG, Jacobsen A, Federspiel B, Bardram L, Krogh A, Lund AH, Friis-Hansen L. microRNA-146a inhibits G protein-coupled receptor-mediated activation of NF-κB by targeting CARD10 and COPS8 in gastric cancer. Mol Cancer. 2012;11:71. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 74] [Cited by in RCA: 88] [Article Influence: 6.3] [Reference Citation Analysis (0)] |
| 16. | Sancho-Albero M, Navascués N, Mendoza G, Sebastián V, Arruebo M, Martín-Duque P, Santamaría J. Exosome origin determines cell targeting and the transfer of therapeutic nanoparticles towards target cells. J Nanobiotechnology. 2019;17:16. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 100] [Cited by in RCA: 188] [Article Influence: 26.9] [Reference Citation Analysis (0)] |
| 17. | Han C, Zhang C, Wang H, Zhao L. Exosome-mediated communication between tumor cells and tumor-associated macrophages: implications for tumor microenvironment. Oncoimmunology. 2021;10:1887552. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 23] [Cited by in RCA: 84] [Article Influence: 16.8] [Reference Citation Analysis (0)] |
| 18. | Zhang R, Bu T, Cao R, Li Z, Wang C, Huang B, Wei M, Yuan L, Yang G. An optimized exosome production strategy for enhanced yield while without sacrificing cargo loading efficiency. J Nanobiotechnology. 2022;20:463. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in RCA: 23] [Reference Citation Analysis (0)] |
| 19. | Ma Y, Dong S, Grippin AJ, Teng L, Lee AS, Kim BYS, Jiang W. Engineering therapeutical extracellular vesicles for clinical translation. Trends Biotechnol. 2025;43:61-82. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 78] [Cited by in RCA: 60] [Article Influence: 60.0] [Reference Citation Analysis (0)] |
| 20. | Erana-Perez Z, Igartua M, Santos-Vizcaino E, Hernandez RM. Genetically engineered loaded extracellular vesicles for drug delivery. Trends Pharmacol Sci. 2024;45:350-365. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 17] [Cited by in RCA: 44] [Article Influence: 22.0] [Reference Citation Analysis (0)] |
| 21. | Gong C, Zhang X, Shi M, Li F, Wang S, Wang Y, Wang Y, Wei W, Ma G. Tumor Exosomes Reprogrammed by Low pH Are Efficient Targeting Vehicles for Smart Drug Delivery and Personalized Therapy against their Homologous Tumor. Adv Sci (Weinh). 2021;8:2002787. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 64] [Cited by in RCA: 80] [Article Influence: 16.0] [Reference Citation Analysis (0)] |
| 22. | Boussadia Z, Lamberti J, Mattei F, Pizzi E, Puglisi R, Zanetti C, Pasquini L, Fratini F, Fantozzi L, Felicetti F, Fecchi K, Raggi C, Sanchez M, D'Atri S, Carè A, Sargiacomo M, Parolini I. Acidic microenvironment plays a key role in human melanoma progression through a sustained exosome mediated transfer of clinically relevant metastatic molecules. J Exp Clin Cancer Res. 2018;37:245. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 135] [Cited by in RCA: 124] [Article Influence: 15.5] [Reference Citation Analysis (0)] |
| 23. | Lee AH, Ghosh D, Quach N, Schroeder D, Dawson MR. Ovarian Cancer Exosomes Trigger Differential Biophysical Response in Tumor-Derived Fibroblasts. Sci Rep. 2020;10:8686. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 8] [Cited by in RCA: 24] [Article Influence: 4.0] [Reference Citation Analysis (0)] |
| 24. | Shukla S, Telraja J, Yadav M, Prakash H. Editorial: Modulation of Macrophage Signaling Pathways During Bacterial Infections. Front Cell Infect Microbiol. 2021;11:689759. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in RCA: 2] [Reference Citation Analysis (0)] |
| 25. | Liu X, Chen W, Zhu G, Yang H, Li W, Luo M, Shu C, Zhou Z. Single-cell RNA sequencing identifies an Il1rn(+)/Trem1(+) macrophage subpopulation as a cellular target for mitigating the progression of thoracic aortic aneurysm and dissection. Cell Discov. 2022;8:11. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 5] [Cited by in RCA: 80] [Article Influence: 20.0] [Reference Citation Analysis (0)] |
| 26. | Ding W, Duan Y, Qu Z, Feng J, Zhang R, Li X, Sun D, Zhang X, Lu Y. Acidic Microenvironment Aggravates the Severity of Hepatic Ischemia/Reperfusion Injury by Modulating M1-Polarization Through Regulating PPAR-γ Signal. Front Immunol. 2021;12:697362. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 23] [Cited by in RCA: 24] [Article Influence: 4.8] [Reference Citation Analysis (0)] |
| 27. | Blaszczak W, Swietach P. What do cellular responses to acidity tell us about cancer? Cancer Metastasis Rev. 2021;40:1159-1176. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 16] [Cited by in RCA: 15] [Article Influence: 3.0] [Reference Citation Analysis (0)] |
| 28. | Coman D, Huang Y, Rao JU, De Feyter HM, Rothman DL, Juchem C, Hyder F. Imaging the intratumoral-peritumoral extracellular pH gradient of gliomas. NMR Biomed. 2016;29:309-319. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 49] [Cited by in RCA: 51] [Article Influence: 5.1] [Reference Citation Analysis (0)] |
| 29. | Lee JW, Guan W, Han S, Hong DK, Kim LS, Kim H. MicroRNA-708-3p mediates metastasis and chemoresistance through inhibition of epithelial-to-mesenchymal transition in breast cancer. Cancer Sci. 2018;109:1404-1413. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 50] [Cited by in RCA: 60] [Article Influence: 7.5] [Reference Citation Analysis (1)] |
| 30. | Li X, Tibenda JJ, Nan Y, Huang SC, Ning N, Chen GQ, Du YH, Yang YT, Meng FD, Yuan L. MiR-204-3p overexpression inhibits gastric carcinoma cell proliferation by inhibiting the MAPK pathway and RIP1/MLK1 necroptosis pathway to promote apoptosis. World J Gastroenterol. 2023;29:4542-4556. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in RCA: 10] [Reference Citation Analysis (0)] |
| 31. | Lin H, Zhou AJ, Zhang JY, Liu SF, Gu JX. MiR-324-5p reduces viability and induces apoptosis in gastric cancer cells through modulating TSPAN8. J Pharm Pharmacol. 2018;70:1513-1520. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 22] [Cited by in RCA: 28] [Article Influence: 3.5] [Reference Citation Analysis (0)] |
| 32. | Etcheverry A, Aubry M, Idbaih A, Vauleon E, Marie Y, Menei P, Boniface R, Figarella-Branger D, Karayan-Tapon L, Quillien V, Sanson M, de Tayrac M, Delattre JY, Mosser J. DGKI methylation status modulates the prognostic value of MGMT in glioblastoma patients treated with combined radio-chemotherapy with temozolomide. PLoS One. 2014;9:e104455. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 16] [Cited by in RCA: 20] [Article Influence: 1.7] [Reference Citation Analysis (0)] |
| 33. | Huang C, Zhao J, Luo C, Zhu Z. Overexpression of DGKI in Gastric Cancer Predicts Poor Prognosis. Front Med (Lausanne). 2020;7:320. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 5] [Cited by in RCA: 11] [Article Influence: 1.8] [Reference Citation Analysis (1)] |
| 34. | Matsson H, Tammimies K, Zucchelli M, Anthoni H, Onkamo P, Nopola-Hemmi J, Lyytinen H, Leppanen PH, Neuhoff N, Warnke A, Schulte-Körne G, Schumacher J, Nöthen MM, Kere J, Peyrard-Janvid M. SNP variations in the 7q33 region containing DGKI are associated with dyslexia in the Finnish and German populations. Behav Genet. 2011;41:134-140. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 21] [Cited by in RCA: 18] [Article Influence: 1.2] [Reference Citation Analysis (0)] |
| 35. | Lin JF, Liu ZX, Chen DL, Huang RZ, Cao F, Yu K, Li T, Mo HY, Sheng H, Liang ZB, Liao K, Han Y, Li SS, Zeng ZL, Gao S, Ju HQ, Xu RH. Nucleus-translocated GCLM promotes chemoresistance in colorectal cancer through a moonlighting function. Nat Commun. 2025;16:263. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 1] [Cited by in RCA: 10] [Article Influence: 10.0] [Reference Citation Analysis (0)] |
| 36. | Gray WD, French KM, Ghosh-Choudhary S, Maxwell JT, Brown ME, Platt MO, Searles CD, Davis ME. Identification of therapeutic covariant microRNA clusters in hypoxia-treated cardiac progenitor cell exosomes using systems biology. Circ Res. 2015;116:255-263. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 342] [Cited by in RCA: 330] [Article Influence: 30.0] [Reference Citation Analysis (0)] |
| 37. | Li C, Fang F, Wang E, Yang H, Yang X, Wang Q, Si L, Zhang Z, Liu X. Engineering extracellular vesicles derived from endothelial cells sheared by laminar flow for anti-atherosclerotic therapy through reprogramming macrophage. Biomaterials. 2025;314:122832. [RCA] [PubMed] [DOI] [Full Text] [Cited by in RCA: 30] [Reference Citation Analysis (0)] |
| 38. | Peng B, Bartkowiak K, Song F, Nissen P, Schlüter H, Siebels B. Hypoxia-Induced Adaptations of N-Glycomes and Proteomes in Breast Cancer Cells and Their Secreted Extracellular Vesicles. Int J Mol Sci. 2024;25:10216. [RCA] [PubMed] [DOI] [Full Text] [Cited by in RCA: 6] [Reference Citation Analysis (0)] |
| 39. | Jain S, Kesharwani P, Tekade RK, Jain NK. One platform comparison of solubilization potential of dendrimer with some solubilizing agents. Drug Dev Ind Pharm. 2015;41:722-727. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 34] [Cited by in RCA: 26] [Article Influence: 2.2] [Reference Citation Analysis (0)] |
| 40. | Fraguas-Sánchez AI, Martín-Sabroso C, Fernández-Carballido A, Torres-Suárez AI. Current status of nanomedicine in the chemotherapy of breast cancer. Cancer Chemother Pharmacol. 2019;84:689-706. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 32] [Cited by in RCA: 53] [Article Influence: 7.6] [Reference Citation Analysis (0)] |
| 41. | Sharifi-Rad J, Quispe C, Patra JK, Singh YD, Panda MK, Das G, Adetunji CO, Michael OS, Sytar O, Polito L, Živković J, Cruz-Martins N, Klimek-Szczykutowicz M, Ekiert H, Choudhary MI, Ayatollahi SA, Tynybekov B, Kobarfard F, Muntean AC, Grozea I, Daştan SD, Butnariu M, Szopa A, Calina D. Paclitaxel: Application in Modern Oncology and Nanomedicine-Based Cancer Therapy. Oxid Med Cell Longev. 2021;2021:3687700. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 160] [Cited by in RCA: 162] [Article Influence: 32.4] [Reference Citation Analysis (0)] |
| 42. | Huis In 't Veld RV, Lara P, Jager MJ, Koning RI, Ossendorp F, Cruz LJ. M1-derived extracellular vesicles enhance photodynamic therapy and promote immunological memory in preclinical models of colon cancer. J Nanobiotechnology. 2022;20:252. [RCA] [PubMed] [DOI] [Full Text] [Full Text (PDF)] [Cited by in Crossref: 2] [Cited by in RCA: 23] [Article Influence: 5.8] [Reference Citation Analysis (0)] |
| 43. | Sharma S, Masud MK, Kaneti YV, Rewatkar P, Koradia A, Hossain MSA, Yamauchi Y, Popat A, Salomon C. Extracellular Vesicle Nanoarchitectonics for Novel Drug Delivery Applications. Small. 2021;17:e2102220. [RCA] [PubMed] [DOI] [Full Text] [Cited by in Crossref: 38] [Cited by in RCA: 77] [Article Influence: 15.4] [Reference Citation Analysis (3)] |
| 44. | Deng C, Zhang H, Li Y, Cheng X, Liu Y, Huang S, Cheng J, Chen H, Shao P, Jiang B, Wang X, Wang K. Exosomes derived from mesenchymal stem cells containing berberine for ulcerative colitis therapy. J Colloid Interface Sci. 2024;671:354-373. [RCA] [PubMed] [DOI] [Full Text] [Cited by in RCA: 24] [Reference Citation Analysis (0)] |