Reduced Tumor Angiogenesis Research Articles

PHPS1 enhances PD-L1 serine phosphorylation by regulating ROS/SHP-2/AMPK activity to promote apoptosis of oral squamous cell carcinoma cells

To investigate the mechanism of PHPS1 for promoting apoptosis of oral squamous cell carcinoma cells and the role of AMPK in regulating tumor angiogenesis under hypoxic conditions. Human oral squamous cell carcinoma Ca9-22 cells cultured in hypoxic conditions (1% O2) were inoculated subcutaneously in 16 nude mice, which were divided into control group and PHPS1 group (n=8) for treatment with 10% DMSO and 10% PHPS1 respectively. Tumor growth in the mice was monitored till 14 days after the treatment, and the xenografts were examined pathologically using HE staining. In Ca9-22 cells cultured in 1% O2, the effect of PHPS1, compound C (an AMPK inhibitor), and their combination on expressions of SHP-2, AMPK, HIF-1α, PD-L1, caspase-8, caspase-3 and BAX were evaluated using Western blotting. In the tumor-bearing nude mice, PHPS1 treatment significantly inhibited tumor growth and neovascularization. HE staining showed significantly reduced tumor angiogenesis in PHPS1-treated mice. In Ca9-22 cells in hypoxic cultures, PHPS1 treatment significantly decreased the expression levels of SHP-2, HIF-1α, PD-L1, ERK2, STAT3 and VEGF and increased the expression of AMPK. The inhibitory effects of PHPS1 on HIF-1α and PD-L1 were obviously attenuated by the addition of compound C. PHPS1 also enhanced the expressions of caspase-3, caspase-8 and Bax proteins and increased the phosphorylation levels of PD-L1 and S195 in Ca9-22 cells, and these effects were effectively attenuated by compound C. PHPS1 can enhance PD-L1 serine phosphorylation by regulating SHP-2/AMPK activity to promote apoptosis of oral squamous cell carcinoma cells under hypoxic conditions.

Natural glyceollin soybean extracts elicited with Aspergillus sojae reduce estrogen-dependent breast cancer growth in orally fed mice.

Previous studies have demonstrated antiestrogenic and antiproliferative effects of these molecules in breast cancer cells. Notably, we have reported that pure synthetic glyceollins I and II act through various pathways, including ERα, FOXM1, AhR, and HIF pathways to inhibit cell proliferation and migration. In this study, the potential antitumor activity of glyceollins enriched in crude soybean extracts, obtained by solid fermentation with Aspergillus sojae, was investigated in vivo on MCF-7 breast cancer cells implanted in the chorioallantoic membrane of the chick egg and on ovariectomized nude mice. The first trial showed a substantial reduction in the migration of MCF-7 cells treated with the natural extracts. However, the natural extracts significantly reduced the estrogen-dependent growth of transplanted tumors in orally fed nude mice. Our results showed that natural soybean extracts slightly but significantly reduced estrogen-dependent growth of the transplanted tumors in orally fed nude mice. These results were confirmed by immunohistochemistry of Ki-67 and histone H3S10 phosphorylation (H3S10P), revealing lower expression of these proliferation markers in the transplanted tumors from mice fed with the fermented extracts. Additionally, compared to the control animals, we observed a lower expression of angiogenesis markers such as CD31 and CD34. Surprisingly, transcriptomic analysis of RNA from transplanted MCF-7 cells revealed no differential gene expression. These results may suggest that orally consumed natural glyceollins exert biological effects throughout the body, acting indirectly to reduce tumor angiogenesis and consequently tumor volume. Overall, our results indicate that glyceollins, elicited components of the soy origin, hold potential therapeutic applications for the prevention and treatment of breast cancer.

Extracellular vesicles derived from dendritic cells loaded with VEGF-A siRNA and doxorubicin reduce glioma angiogenesis in vitro

BackgroundNumerous attempts have been devoted to designing anti-angiogenic agents as a strategy to slow tumor growth and progression. Clinical applications of conventional anti-angiogenic agents face some challenges, e.g., off-target effects for TKIs and also low solid tumor penetration for mAbs. Furthermore, although anti-angiogenic therapy provides a normalization window for better chemo-RT response, in long-term treatments, tumor hypoxia as a result of total removal of VEGF-A by mAbs from the TME or complete blockade of TK receptors induces over-activation of compensatory angiogenic pathways, causing escape. Herein, we investigate the efficacy of si-DOX-DC-EVs to reduce glioma angiogenesis and invasiveness. MethodsMature DCs were generated from PBMC and EVs were isolated from the DCs culture media. siRNA and Doxorubicin were loaded into EVs by EP and incubation. Afterward, the uptake of DC-EVs was assessed by flow cytometry, and the subcellular localization of EVs was tested by confocal imaging. Tube formation assay was performed to assess the efficacy of si-DOX-DC-EVs to reduce tumor angiogenesis which was analyzed by DHM. Morphometric analysis of apoptotic cells was performed by DHM and confocal imaging and further, ELISA was performed for hypoxia-related and angiogenic cytokines. The impact of our theranostic system “si-DOX-DC-MVs” on the formation of vascular mimics, colonies, and invasion of C6 cells was checked in vitro. Afterward, orthotropic rat models of glioma were generated and the optimal administration route was selected by in vivo fluorescent analysis. Then, the microvessel density, vimentin expression, and accumulation of immune cells in tumoral tissues were assessed by IHC. Finally, necropsy and autopsy analyses were performed to check the safety of our theranostic agent. ResultsDC-EVs loaded with si-DOX-DC-EVs were successfully uptaken by cells with different subcellular trafficking for MVs and exosomes, reduced tumor angiogenesis in DHM analysis, and induced apoptosis in tumoral cells. Moreover, using DHM, we performed a detailed label-free analysis of tip cells which suggested that the tip cells in si-DC-MV treatments lost their geometrical migration capacity to form tube-like structures. Furthermore, the ELISAs performed highlighted that there is a mild overactivation of compensatory Tie2/Ang2 pathway after VEGF-A blockade which confers with severe hypoxia and sustains normal angiogenesis which is the optimal goal of anti-angiogenesis therapy for cancer to avoid resistance.The results of our VM analyses indicated that si-DOX-DC-MVs completely inhibited VM process. Moreover, the invasion, migration, and colony formation of the C6 cells treated with si-DOX-MVs were the least among all treatments. IN was the optimal route of administration. The MVD analyses indicated that si-DOX-DC-MVs reduced the number of tumoral microvessels and normalized vessel morphology. Intense CD8+ T cells were observed near the tumoral vessels in the si-DOX-DC-MVs group and with minimal activation of MT (low Vimentin expression). Necropsy and toxicology results proved that the theranostic system proposed is safe. ConclusionsDC-EVs loaded with VEGF-A siRNA and Doxorubicin were more potent than BV alone as a multi-disciplinary strategy that combats glioma growth by cytotoxic impacts of DOX and inhibits angiogenesis by VEGF-A siRNAs with excess immunologic benefits from DC-EVs. This next-generation anti-angiogenic agent normalizes tumor vessel density rather than extensively eliminating tumor vessels causing hypoxia and mesenchymal transition.

Abstract 2786: Seizure related 6 homolog (SEZ6), a cancer stem cell-specific secreted protein, is a novel GBM therapeutic target

Abstract Cancer Stem Cells (CSCs) are known to initiate a tumor, resist chemotherapy and radiotherapy, and also cause tumor recurrence. A deeper understanding of CSC biology is needed for a more effective therapeutic response. An integrative study of regulated transcriptome of human patient-derived Glioma Stem-like Cells (GSCs) using Gene Ontology (GO) analysis and Gene Set Enrichment Analysis (GSEA) identified a potential role for GSC in tumor angiogenesis. We chose Seizure Related 6 Homolog (SEZ6), one of the Wnt target genes, for further study, as its downregulation inhibited the potential of GSCs to induce angiogenesis without impacting GSC growth. SEZ6, a transmembrane protein, is shed upon cleavage by Beta-site APP cleaving enzyme 1 (BACE 1) and is known to be involved in normal neuronal functions; however, its role in cancer has not been studied yet. SEZ6 silencing inhibited GSC-initiated tumor growth in an intracranial orthotopic mouse model due to reduced tumor angiogenesis and increased the mouse survival. Pre-treatment with a pharmacological inhibitor of BACE1 (BACE1i) prevented SEZ6 shedding from GSCs and inhibited the ability of the GSC-secretome to induce angiogenesis. Further investigations are ongoing to establish the use of BACE1i or BACE1-silencing as a novel therapeutic strategy to inhibit SEZ6-mediated tumor angiogenesis. We also carried out a protein array-based investigation, which revealed that SEZ6 induced IL8 in endothelial cells to induce angiogenesis. Additional investigation showed that SEZ6 activated the TGFβ pathway in endothelial cells to induce IL8. SEZ6 interacted with TGFβRII as seen by confocal microscopy and co-immunoprecipitation. Generation of various deletion mutants of SEZ6 revealed that three Sushi [also known as complement control protein (CCP) module or the short consensus repeat (SCR)] domains, 3, 4, and 5, interacted with TGFβRII. Molecular docking and dynamic simulation studies identified Sushi-3 domain as the mediator of the interaction between SEZ6 and TGFβRII extracellular domain. Based on the above results, experiments are underway to design specific inhibitors of this interaction. We are also developing an improved BACE1 inhibitor that would inhibit the shedding of SEZ6 into the extracellular milieu to inhibit SEZ6 functions. Thus, this study establishes that GSC-secreted SEZ6 activates the TGFβ pathway in endothelial cells to induce tumor angiogenesis and proposes a novel therapeutic target. Citation Format: Arani Mukherjee, Shreoshi Sengupta, Abhishek Chowdhury, Anand Srivastava, Kumar Somasundaram. Seizure related 6 homolog (SEZ6), a cancer stem cell-specific secreted protein, is a novel GBM therapeutic target [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2786.

The Use of Isoflavones as Lung Cancer Chemoprevention Agents and their Implications in Treatment through Radio Sensitization.

Epidemiological trends in cancer research show that lung cancer can affect up to 1 in 15 men and 1 in 17 women. With incidence rates as high as these and significant associated mortality and morbidity, it is no wonder that lung cancer is one of the main areas of research focused on cancer. Advances in targeted treatments and specialized irradiation protocols have allowed the treatment of more advanced cases. However, as the patient numbers grow, so does the need for cancer-preventive strategies. The present narrative review focuses on soy isoflavones' role in the chemoprevention of lung cancer and their possible role in therapeutic adjuncts. Laboratory studies on lung cancer cell lines have shown that isoflavones can induce apoptosis, tamper with the expression of proliferative molecular pathways, and even reduce tumor angiogenesis. Additionally, population-level studies have emerged that correlate the consumption of isoflavonoids with reduced risk for the development of lung cancer. Interestingly enough, the literature also contains small-scale studies with evidence of isoflavones being effective chemotherapeutic adjuncts that are currently understudied. Our literature review underlines such findings and provides a call for the enhancement of research regarding naturally occurring dietary products with possible anticarcinogenic effects.

Vasohibin-2-Targeting Therapies for the Treatment of Pancreatic Ductal Adenocarcinoma

As pancreatic ductal adenocarcinoma (PDAC) is extremely malignant and refractory, therapeutic options for this cancer are anticipated worldwide. We isolated vasohihibin-2 (VASH2) and observed its overexpression in various types of cancer. We then noticed that upregulated expression of VASH2 in patients with PDAC resulted in a conspicuous reduction in the postoperative survival period and further revealed that the abrogation of Vash2 expression in pancreatic cancer cells inhibited its growth and metastasis and augmented tumor infiltration of CD8+ cells in the mouse model. We developed VASH2-targeting therapies, 2',4'-BNA-based antisense oligonucleotide targeting VASH2 (VASH2-ASO) as a nucleotide-based therapy, and VASH2-peptide vaccine as an antibody-based therapy. We also showed that the VASH2-peptide vaccine inhibited PDAC metastasis in an orthotopic mouse model. Here, we expanded our analysis of the efficacy of VASH2-targeting therapies for PDAC. VASH2-ASO treatment inhibited the growth of primary tumors by reducing tumor angiogenesis, normalizing tumor vessels, preventing ascites accumulation and distant metastasis to the liver and lungs, and augmenting the infiltration of CD8+ cells in metastatic tumors. VASH2-peptide vaccine did not affect the infiltration of CD8+ cells into tumors. The present study revealed that VASH2-targeting therapies are promising options for the treatment of PDAC. VASH2-ASO therapy can be administered at any stage of PDAC. Because of the increase of CD8+ cell infiltration, the combination therapy with immune checkpoint inhibitors may be an attractive option. The VASH2-peptide vaccine therapy may be useful for preventing metastasis and/or recurrence after successful initial treatment.

Inhibition of SUV39H1 reduces tumor angiogenesis via Notch1 in oral squamous cell carcinoma.

Targeting tumor angiogenesis is an important approach in advanced tumor therapy. Here we investigated the effect of the suppressor of variegation 3-9 homolog 1 (SUV39H1) on tumor angiogenesis in oral squamous cell carcinoma (OSCC). The GEPIA database was used to analyze the expression of SUV39H1 in various cancer tissues. The expression of SUV39H1 in OSCC was detected by immunohistochemistry, and the correlation between SUV39H1 and Notch1 and microvascular density (MVD) was analyzed. The effect of SUV39H1 inhibition on OSCC was investigated in vivo by chaetocin treatment. The migration and tube formation of vascular endothelial cells by conditioned culture-medium of different treatments of oral squamous cell cells were measured. The transcriptional level of SUV39H1 is elevated in various cancer tissues. The transcription level of SUV39H1 in head and neck squamous cell carcinoma was significantly higher than that in control. Immunohistochemistry result showed increased SUV39H1 expression in OSCC, which was significantly correlated with T staging. The expression of SUV39H1 was significantly correlated with Notch1 and CD31. In vivo experiment chaetocin treatment significantly inhibit the growth of tumor, and reduce SUV39H1, Notch1, CD31 expression. The decreased expression of SUV39H1 in OSCC cells lead to the decreased expression of Notch1 and VEGF proteins, as well as the decreased migration and tube formation ability of vascular endothelial cells. Inhibition of Notch1 further enhance this effect. Our results suggest inhibition of SUV39H1 may affect angiogenesis by regulating Notch1 expression. This study provides a foundation for SUV39H1 as a potential therapeutic target for OSCC.

GPR4 Knockout Attenuates Intestinal Inflammation and Forestalls the Development of Colitis-Associated Colorectal Cancer in Murine Models.

GPR4 is a proton-sensing G protein-coupled receptor highly expressed in vascular endothelial cells and has been shown to potentiate intestinal inflammation in murine colitis models. Herein, we evaluated the proinflammatory role of GPR4 in the development of colitis-associated colorectal cancer (CAC) using the dextran sulfate sodium (DSS) and azoxymethane (AOM) mouse models in wild-type and GPR4 knockout mice. We found that GPR4 contributed to chronic intestinal inflammation and heightened DSS/AOM-induced intestinal tumor burden. Tumor blood vessel density was markedly reduced in mice deficient in GPR4, which correlated with increased tumor necrosis and reduced tumor cell proliferation. These data demonstrate that GPR4 ablation alleviates intestinal inflammation and reduces tumor angiogenesis, development, and progression in the AOM/DSS mouse model.

Disintegrin-like Protein Strategy to Inhibit Aggressive Triple-Negative Breast Cancer.

Venoms are a rich source of bioactive compounds, and among them is leberagin-C (Leb-C), a disintegrin-like protein derived from the venom of Macrovipera lebetina transmediterrannea snakes. Leb-C has shown promising inhibitory effects on platelet aggregation. Previous studies have demonstrated that this SECD protein specifically targets α5β1, αvβ3, and αvβ6 integrins through a mimic mechanism of RGD disintegrins. In our current study, we focused on exploring the potential effects of Leb-C on metastatic breast cancer. Our findings revealed that Leb-C disrupted the adhesion, migration, and invasion capabilities of MDA-MB-231 breast cancer cells and its highly metastatic D3H2LN sub-population. Additionally, we observed significant suppression of adhesion, migration, and invasion of human umbilical vein endothelial cells (HUVECs). Furthermore, Leb-C demonstrated a strong inhibitory effect on fibroblast-growth-factor-2-induced proliferation of HUVEC. We conducted in vivo experiments using nude mice and found that treatment with 2 µM of Leb-C resulted in a remarkable 73% reduction in D3H2LN xenograft tumor size. Additionally, quantification of intratumor microvessels revealed a 50% reduction in tumor angiogenesis in xenograft after 21 days of twice-weekly treatment with 2 µM of Leb-C. Collectively, these findings suggest the potential utility of this disintegrin-like protein for inhibiting aggressive and resistant metastatic breast cancer.

The 14-Kilodalton Human Growth Hormone Fragment a Potent Inhibitor of Angiogenesis and Tumor Metastasis.

The 14-kilodalton human growth hormone (14 kDa hGH) N-terminal fragment derived from the proteolytic cleavage of its full-length counterpart has been shown to sustain antiangiogenic potentials. This study investigated the antitumoral and antimetastatic effects of 14 kDa hGH on B16-F10 murine melanoma cells. B16-F10 murine melanoma cells transfected with 14 kDa hGH expression vectors showed a significant reduction in cellular proliferation and migration associated with an increase in cell apoptosis in vitro. In vivo, 14 kDa hGH mitigated tumor growth and metastasis of B16-F10 cells and was associated with a significant reduction in tumor angiogenesis. Similarly, 14 kDa hGH expression reduced human brain microvascular endothelial (HBME) cell proliferation, migration, and tube formation abilities and triggered apoptosis in vitro. The antiangiogenic effects of 14 kDa hGH on HBME cells were abolished when we stably downregulated plasminogen activator inhibitor-1 (PAI-1) expression in vitro. In this study, we showed the potential anticancer role of 14 kDa hGH, its ability to inhibit primary tumor growth and metastasis establishment, and the possible involvement of PAI-1 in promoting its antiangiogenic effects. Therefore, these results suggest that the 14 kDa hGH fragment can be used as a therapeutic molecule to inhibit angiogenesis and cancer progression.

CLINICAL RESEARCH PROGRESS OF COMPOUND KUSHEN INJECTION ON ANTI-TUMOR EFFECT

Kushen, a traditional Chinese medicine, has the functions of clearing away heat and dampness, reducing inflammation and detoxifying, and its alkaloids are its main anti-tumor substances. In recent years, as the extract of Kushen, compound Kushen injection has the characteristics of significant anti-tumor efficacy and few side effects, it is often combined with various anti-tumor treatments such as surgery, chemotherapy, radiotherapy, and targeted therapy in clinical practice in China. This paper summarizes the animal and clinical experiments of cervical, breast, gastric, and colon cancer through the drug effect of the compound Kushen injection. Many studies have shown that compound Kushen injection mainly exerts its anti-tumor effect by regulating tumor cell proliferation, inducing tumor cell apoptosis, reducing tumor angiogenesis, and improving body immunity. Based on traditional anti-tumor therapy, adding the compound Kushen injection can have a more apparent therapeutic effect on patients, reduce the side effects of anti-tumor therapy, improve patients' quality of life, and prolong patients' survival time. This paper significantly summarizes the research on the anti-tumor treatment of compound Kushen injection, further clarifies the anti-tumor effect of the effective extracts of traditional Chinese medicine, which provides a basis for future anti-tumor research of compound Kushen injection and a basis for the clinical promotion of the drug.

Abstract 571: Angiopoietin-like 4 as a potential therapeutic target for clear cell renal carcinoma

Abstract Renal cell carcinoma (RCC) is the 7th most common type of cancer, with over 400,000 new cases each year and 170,000 deaths worldwide. RCCs are grouped into several histological subtypes, with clear cell renal cell carcinoma (ccRCC) being the most common and most aggressive type. The first line of therapy for ccRCC is tyrosine kinase inhibitors (TKIs) which target multiple kinases including the VEGF receptor. The treatment can be used alone or in combination with immune checkpoint inhibitors. However, most patients develop resistance to the TKIs within six months of treatment. Therefore, our aim is to develop novel single or combinatorial therapies and discover ways to overcome the resistance to TKIs. By analyzing TCGA and published datasets from GEO, we found that angiopoietin-like 4 (ANGPTL4) is significantly elevated in both the WT and TKI-resistant ccRCC tumors. ANGPTL4 has been shown to participate in angiogenesis in several other tumor models, and blocking ANGPTL4 with an antibody in vitro, reduces the angiogenic potential of 786-O and RCC4 human ccRCC cell lines, suggesting a pro-angiogenic effect of ANGPTL4 in ccRCC. Knockout of ANGPTL4 in Caki-1 xenograft tumor model significantly reduced tumor angiogenesis and increased vessel normalization, as evidenced by increased expression of VEGFR2 and adhesion molecules on endothelial cells. The increased normalization of blood vessels is an indication of the therapeutic potential of combing anti-ANGPTL4 therapy with immune checkpoint inhibitors. Furthermore, both knockout of ANGPTL4 in 786-O cells and treatment of 786-O tumor-bearing mice with an ANGPTL4 blocking antibody significantly reduced tumor growth. To illustrate the mechanism of ANGPTL4-induced angiogenesis, we performed live cell receptor capturing to find cANGPTL4 binding partners on the cell membrane of human endothelial cells. After the mass spectrometry analysis, several novel receptors were found, and the top candidate is endothelial cell surface chemotaxis receptor (ECSCR), which is highly expressed on endothelial and adipocytes. Previous studies have shown that ECSCR can potentiate angiogenic signaling, which is similar to our data showing that ANGPTL4 can synergistically increase FGF2-induced angiogenesis. Thus, ANGPTL4 may promote angiogenesis in ccRCC by potentiating other angiogenic signaling via interaction with ECSCR. To summarize, we demonstrated the pro-angiogenic potential of ANGPTL4 in ccRCC and its critical role in tumor development. Knockout or antibody-mediated inhibition of ANGPTL4 reduced tumor angiogenesis and growth and increased vessel normalization. This data supports the therapeutic potential of ANGPTL4 targeted treatment in ccRCC. Citation Format: Zeng Jin, Umasankar De, Lei Wang, Jeremy Kleberg, Weizhou Zhang, Ryan Kolb. Angiopoietin-like 4 as a potential therapeutic target for clear cell renal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 571.

Combination of immune checkpoint blockade and targeted gene regulation of angiogenesis for facilitating antitumor immunotherapy.

The rapid development of tumor immunotherapy has improved the management of patients with cancer. However, several key problems of tumor immunotherapy, including the insufficient activation of effector T cells, poor tumor invasion, and poor immune killing ability, lead to a low response rate. In the present study, a synergistic strategy was developed by combining in situ tumor vaccines, gene-mediated downregulation of tumor angiogenesis, and anti-PD-L1 therapy. In situ tumor vaccines and antitumor angiogenesis were achieved by codelivering unmethylated cytosine-phosphate-guanine (CpG) and vascular endothelial growth factor (VEGF)-silencing gene (shVEGF) via a hyaluronic acid (HA)-modified HA/PEI/shVEGF/CpG system. Necrotic tumor cells and CpG adjuvants formed in situ tumor vaccines and activated the host immune response. Moreover, VEGF silencing reduced tumor angiogenesis and prompted the homogeneous distribution of tumor blood vessels to facilitate immune cell infiltration. Meanwhile, anti-angiogenesis also improved the immunosuppressive tumor microenvironment. To further improve the specific tumor-killing effect, an anti-PD-L1 antibody was introduced for immune checkpoint blockade, thereby boosting antitumor immune responses. The combination therapy strategy presented in the present study could act in the multiple stages of the tumor immunotherapy cycle, which is expected to offer a new avenue for clinical tumor immunotherapy.

A zebrafish HCT116 xenograft model to predict anandamide outcomes on colorectal cancer

Colon cancer is one of the leading causes of death worldwide. In recent years, cannabinoids have been extensively studied for their potential anticancer effects and symptom management. Several in vitro studies reported anandamide’s (AEA) ability to block cancer cell proliferation and migration, but evidence from in vivo studies is still lacking. Thus, in this study, the effects of AEA exposure in zebrafish embryos transplanted with HCT116 cells were evaluated. Totally, 48 hpf xenografts were exposed to 10 nM AEA, 10 nM AM251, one of the cannabinoid 1 receptor (CB1) antagonist/inverse agonists, and to AEA + AM251, to verify the specific effect of AEA treatment. AEA efficacy was evaluated by confocal microscopy, which demonstrated that these xenografts presented a smaller tumor size, reduced tumor angiogenesis, and lacked micrometastasis formation. To gain deeper evidence into AEA action, microscopic observations were completed by molecular analyses. RNA seq performed on zebrafish transcriptome reported the downregulation of genes involved in cell proliferation, angiogenesis, and the immune system. Conversely, HCT116 cell transcripts resulted not affected by AEA treatment. In vitro HCT116 culture, in fact, confirmed that AEA exposure did not affect cell proliferation and viability, thus suggesting that the reduced tumor size mainly depends on direct effects on the fish rather than on the transplanted cancer cells. AEA reduced cell proliferation and tumor angiogenesis, as suggested by socs3 and pcnp mRNAs and Vegfc protein levels, and exerted anti-inflammatory activity, as indicated by the reduction of il-11a, mhc1uba, and csf3b mRNA. Of note, are the results obtained in groups exposed to AM251, which presence nullifies AEA’s beneficial effects. In conclusion, this study promotes the efficacy of AEA in personalized cancer therapy, as suggested by its ability to drive tumor growth and metastasis, and strongly supports the use of zebrafish xenograft as an emerging model platform for cancer studies.

Clathrin light chain-conjugated drug delivery for cancer.

Targeted drug delivery systems hold the remarkable potential to improve the therapeutic index of anticancer medications markedly. Here, we report a targeted delivery platform for cancer treatment using clathrin light chain (CLC)-conjugated drugs. We conjugated CLC to paclitaxel (PTX) through a glutaric anhydride at high efficiency. Labeled CLCs localized to 4T1 tumors implanted in mice, and conjugation of PTX to CLC enhanced its delivery to these tumors. Treatment of three different mouse models of cancer-melanoma, breast cancer, and lung cancer-with CLC-PTX resulted in significant growth inhibition of both the primary tumor and metastatic lesions, as compared to treatment with free PTX. CLC-PTX treatment caused a marked increase in apoptosis of tumor cells and reduction of tumor angiogenesis. Our data suggested HSP70 as a binding partner for CLC. Our study demonstrates that CLC-based drug-conjugates constitute a novel drug delivery platform that can augment the effects of chemotherapeutics in treating a variety of cancers. Moreover, conjugation of therapeutics with CLC may be used as means by which drugs are delivered specifically to primary tumors and metastatic lesions, thereby prolonging the survival of cancer patients.

Visfatin-Induced Inhibition of miR-1264 Facilitates PDGF-C Synthesis in Chondrosarcoma Cells and Enhances Endothelial Progenitor Cell Angiogenesis.

New treatments for chondrosarcoma are extremely important. Chondrosarcoma is a primary malignant bone tumor with a very unfavorable prognosis. High-grade chondrosarcoma has a high potential to metastasize to any organ in the body. Platelet-derived growth factor (PDGF) is a potent angiogenic factor that promotes tumor angiogenesis and metastasis. The adipocytokine visfatin promotes metastatic potential of chondrosarcoma; however, the role of visfatin in angiogenesis in human chondrosarcoma is unclear. We report that the levels of PDGF-C expression were positively correlated with tumor stages, significantly higher than the levels of expression in normal cartilage. Visfatin increased PDGF-C expression and endothelial progenitor cell (EPC) angiogenesis through the PI3K/Akt/mTOR signaling pathway, and dose-dependently down-regulated the synthesis of miR-1264, which targets the 3'-UTR of PDGF-C. Additionally, we discovered inhibition of visfatin or PDGF-C in chondrosarcoma tumors significantly reduced tumor angiogenesis and size. Our results indicate that visfatin inhibits miR-1264 production through the PI3K/Akt/mTOR signaling cascade, and thereby promotes PDGF-C expression and chondrosarcoma angiogenesis. Visfatin may be worth targeting in the treatment of chondrosarcoma angiogenesis.

Research Progress of Xiaoaiping Injection on Anti-tumor

Xiaoaiping injection is an injection solution of effective substances extracted from the traditional Chinese medicine Tongguanteng, which has the effects of reducing inflammation and phlegm, clearing heat, and detoxifying. Xiaoaiping injection has no apparent side effects, and the price of Xiaoaiping injection is low. Xiaoaiping injection is commonly used in clinical anti-tumor therapy in China and is often combined with tumor therapy such as surgery, chemotherapy, radiotherapy, and targeted therapy, and it has an excellent curative effect. Doctors and patients widely praise it. This paper summarizes the therapeutic effect of Xiaoaiping injection in treating breast cancer, liver cancer, lung cancer, and esophageal cancer in clinical and animal experiments. Many studies have supported that Xiaoaiping injection can regulate the proliferation and apoptosis of tumor cells, reduce tumor angiogenesis, and improve human immunity to achieve anti-tumor effects. During combined chemotherapy treatment, Xiaoaiping injection can also alleviate the adverse reactions caused by chemotherapy and significantly improve patient's quality of life. This paper summarizes the research on the anti-tumor therapy of Xiaoaiping injection, which provides a basis for the clinical promotion of the drug and assistance in developing traditional Chinese medicine.

Synergy of Polydopamine Nanovaccine and Endostar Alginate Hydrogel for Improving Antitumor Immune Responses Against Colon Tumor.

BackgroundTumor immunotherapy, a novel type of therapeutic treatment, has a wide range of applications with potentially prolonged benefits. However, current immunotherapy has a low overall response rate in treating a variety of tumors. Combination of immunotherapy with other therapies can improve the therapeutic response rates. The purpose of this work was to explore the potential of anti-angiogenic treatment in combination with tumor cell lysate loaded polydopamine nanoparticle vaccine as a therapeutic strategy for colon tumor.MethodsWe grafted tumor cell lysate onto polydopamine nanoparticles as nano-vaccine (TCLN) and fabricated alginate hydrogel loaded with Endostar (EH), then detected characteristics of EH and TCLN. We also estimated the cytotoxicity of EH/TCLN in vitro. In the tumor-bearing mouse model, we evaluated the antitumor effect of EH/TCLN treatment, and developed the animal survival study. After performing the EH/TCLN treatment, we also analyzed T cells and DCs using flow cytometry, and determined T cell responses and tumor microenvironmental cytokines. At last, we assessed the effect of the EH/TCLN treatment on anti-angiogenesis further.ResultsWhen applied in combination with TCLN in MC-38 tumor-bearing mice, EH/TCLN significantly suppressed tumor growth with more than half of the mice showing tumor regression. In addition, EH/TCLN treatment resulted in noticeable changes in the tumor microenvironment. As compared with the control group, EH/TCLN treatment led to significantly reduced tumor angiogenesis and expression of tumor microenvironment-related cytokines (TMCs), increased proportion of CD8+ T cells in the spleen, lymph node and tumor, elevated activity of cytotoxic T lymphocytes (CTLs) and tumor cell apoptosis.ConclusionThe present study demonstrated that the EH/TCLN treatment effectively created a favorable immune microenvironment for the induction of antitumor immunity and improved antitumor immune responses.

Roles of CD4+ T cells as mediators of antitumor immunity.

It has been well established that CD8+ T cells serve as effector cells of the adaptive immune response against tumors, whereas CD4+ T cells either help or suppress the generation of CD8+ cytotoxic T cells. However, in several experimental models as well as in cancer patients, it has been shown that CD4+ T cells can also mediate antitumor immunity either directly by killing tumor cells or indirectly by activating innate immune cells or by reducing tumor angiogenesis. In this review, we discuss the growing evidence of this underappreciated role of CD4+ T cells as mediators of antitumor immunity.

Lenvatinib for effectively treating antiangiogenic drug-resistant nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) clinical trials show that antiangiogenic drugs (AADs) fail to achieve the expected efficacy, and combining AAD with chemoradiotherapy does not show superiority over chemoradiotherapy alone. Accumulating evidence suggests the intrinsic AAD resistance in NPC patients with poorly understood molecular mechanisms. Here, we describe NPC-specific FGF-2 expression-triggered, VEGF-independent angiogenesis as a mechanism of AAD resistance. Angiogenic factors screening between AAD-sensitive cancer type and AAD-resistant NPC showed high FGF-2 expression in NPC in both xenograft models and clinical samples. Mechanistically, the FGF-2-FGFR1-MYC axis drove endothelial cell survival and proliferation as an alternative to VEGF-VEGFR2-MYC signaling. Genetic knockdown of FGF-2 in NPC tumor cells reduced tumor angiogenesis, enhanced AAD sensitivity, and reduced pulmonary metastasis. Moreover, lenvatinib, an FDA recently approved multi-kinase inhibitor targeting both VEGFR2 and FGFR1, effectively inhibits the tumor vasculature, and exhibited robust anti-tumor effects in NPC-bearing nude mice and humanized mice compared with an agent equivalent to bevacizumab. These findings provide mechanistic insights on FGF-2 signaling in the modulation of VEGF pathway activation in the NPC microenvironment and propose an effective NPC-targeted therapy by using a clinically available drug.