Vascular Endothelial Growth Factor Expression Research Articles

LncRNA H19 Promotes Angiogenesis in Mouse Pulmonary Artery Endothelial Cells by Regulating the HIF-1α/VEGF Signaling Pathway.

Persistent pulmonary hypertension of the newborn (PPHN) is a syndrome of acute respiratory failure characterized by systemic hypoxemia and elevated pulmonary arterial pressure, which leads to pathological changes in pulmonary vascular remodeling and endothelial cell function. Long non-coding RNA (lncRNA) H19 has been shown to be involved in the regulation of arterial endothelial cell function, but its regulatory role in PPHN is not fully understood. In the present study, mouse pulmonary artery endothelial cells (MPAECs) were cultured in a hypoxic conditions. Subsequently, the regulatory function of lncRNA H19 on MPAECs was explored by constructing adenoviruses knocking down and overexpressing lncRNA H19. The results revealed that the hypoxic conditions could induce the proliferation and migration of MPAECs, as well as the high expression of lncRNA H19 in MPAECs. Knockdown of lncRNA H19 expression in MPAECs reversed hypoxic environment-induced functional changes in endothelial cells, whereas overexpression of lncRNA H19 further enhanced the proliferation and migration of MPAECs. In addition, lncRNA H19 upregulated the hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF) pathway through sponge of miNA-20a-5p, which in turn promoted changes in endothelial cell function. LncRNA H19 may interfere with vascular remodeling in hypoxia-induced pulmonary hypertension by upregulating the expression of HIF-1α and VEGF in vascular endothelial cells.

Adipose-mesenchymal stem cell-derived extracellular vesicles enhance angiogenesis and skin wound healing via bFGF-mediated VEGF expression.

This study aimed to investigate whether extracellular vesicles (EVs) derived from adipose-derived mesenchymal stem cells (ASCs) promote skin wound healing by delivering basic fibroblast growth factor (bFGF) to enhance vascular endothelial growth factor (VEGF) expression. ASCs were isolated and transfected with either a bFGF knockdown lentivirus (Lv-sh-bFGF) or a control lentivirus (Lv-sh-NC). EVs were extracted from ASCs cultures and characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting for surface markers. EVs were extracted from the conditioned mediums of ASCs and subjected to different treatments. These EVs or control treatments were injected at the wound edges. Wound healing was assessed using histological techniques, including H&E and Masson's trichrome staining to evaluate tissue regeneration, collagen organization, and immunohistochemistry for CD31 to quantify microvessel density. Protein expression of bFGF and VEGF was measured by Western blotting. ASC-derived EVs significantly promoted angiogenesis and improved skin wound healing. EVs encapsulating bFGF enhanced VEGF expression in the wound tissue, while knockdown of bFGF reduced both bFGF and VEGF expression, leading to delayed wound healing. Further knockdown of VEGF partially reversed the pro-angiogenic and wound-healing effects of bFGF-encapsulated EVs. This study demonstrates that ASC-derived EVs promoted skin wound repair by enhancing angiogenesis and accelerating tissue regeneration through the bFGF/VEGF axis. These findings highlight the therapeutic potential of ASCs-derived EVs in improving skin wound healing.

Inhibition of Acute Myeloid Leukaemia Development by Bittersweet Based on miR-let-7a Regulating Vascular Endothelial Growth Factor A Resistance Mechanism

Acute myeloid leukaemia (AML) is closely related to regulation of miR-let-7a and vascular endothelial growth factor A (VEGFA). Picrasidine is a traditional Chinese medicine extract with antitumour effects, but its mechanism of action in AML is unclear. This study investigated picloram’s effect on AML and its relationship with miR-let-7a regulation of VEGFA resistance mechanism. Bone marrow samples from leukaemia patients in the Department of Haematology of our hospital were collected, and RT-PCR detected miR-let-7a and VEGFA expression in the bone marrow of healthy individuals and leukaemia patients. At the same time, cell culture of AML-resistant cell line K562/ADM was performed, which was divided into NC group, Picrasidine L group, Picrasidine M group, Picrasidine H group, si-NC group, Picrasidine H+miR-let-7a inhibitor group, Picrasidine H+miR-let-7a mimic group, miR-let-7a mimic+hVEGF-IN-1 group, miR-let-7a inhibitor+hVEGF-IN-1 group, and Picrasidine H+miR-let-7a mimic+hVEGF-IN-1 group. Cell proliferation and apoptosis was detected and correlation between miR-let-7a and VEGFA was analyzed by clinical samples. Picrasidine had a significant ameliorative effect on acute myeloid leukaemia in a dose-dependent manner. miR-let-7a was lowly expressed and VEGFA was highly expressed in AML patients. miR-let-7a and VEGFA showed significant correlation in human AML disease staging, and there was a statistically significant difference (p <0.05). That is to say, picloram promotes miR-let-7a expression, thus achieving inhibition of VEGFA, which in turn promotes apoptosis of AML drug-resistant cell line K562/ADM and inhibits its proliferation. The ameliorative effect of Picrasidine on acute myeloid leukaemia was achieved by upregulating miR-let-7a and downregulating VEGFA.

Effect of Jinfeng Pill on ovarian function of Tripterygium wilfordii polyglycoside tablets induced poor ovarian response rats

Objective: This study aimed to assess the effects of the Jinfeng pill on ovarian volume and endocrine function in rats with poor ovarian response and to elucidate the gene expression changes that contribute to the Jinfeng pill’s role in improving ovarian function. Methods: Thirty 8-week-old specific pathogen-free (SPF) female rats were randomly assigned to five groups: control, model, estradiol, Jinfeng pill, and Jinfeng pill + estradiol, with six rats in each group. All groups, except the control were administered Tripterygium glycoside tablets. After establishing the poor ovarian response model, the three intervention groups (estradiol, Jinfeng pill, and Jinfeng pill + estradiol) received their respective drug interventions. Hematoxylin-eosin staining was employed to assess follicle distribution, and immunohistochemistry was used to evaluate the expression of vascular endothelial growth factor and endostatin in rat ovaries. Hormone levels were measured using ELISA. Results: The uterine and total ovarian masses in the three intervention groups were higher than those in the model group; however, the differences were not statistically significant. The ovaries of the Jinfeng pill and Jinfeng pill + estradiol groups contained more follicles at various developmental stages. The three intervention groups showed significantly increased vascular endothelial growth factor expression and decreased endostatin expression compared with the model group, with the Jinfeng pill + estradiol group exhibiting the most pronounced differences. Anti-Müllerian hormone levels were slightly elevated in the Jinfeng pill group compared to the model group. However, only the Jinfeng pill + estradiol group showed a statistically significant decrease in follicle-stimulating hormone levels compared to the model group (P = 0.01). Conclusions: Jinfeng pill modulates ovarian protein expression, increases ovarian volume, and improves blood circulation. Additionally, it may elevate hormone levels and enhance ovarian response function in rats with poor ovarian function.

Mebendazole nano-medication which incorporation nano-chitosan decorated by zinc oxide nanoparticles for intestinal murine trichinellosis through a drug delivery system

The objective is to develop the drug system of mebendazole (MBZ) as an antiparasitic drug via the self-assembly of MBZ with nano chitosan (Cs) and zinc oxide nanoparticles (ZnO NPs). The method of nanocomposite was fabricated by the precipitation method. Sixty male Swiss albino mice (25–30 g) were used, equally divided into six groups; group 1 (G1) included the negative control, and the other five groups (G2, G3, G4, G5, and G6) were infected with 200 Trichinella spiralis (T. spiralis) larvae. G2 refers to the positive control (infected without treatment), G3 is treated with 200 mg/kg/day of MBZ, while G4, G5, and G6 were treated with Cs@MBZ (400 mg/kg/day), Cs-MBZ@ZnO full dose of MBZ, and Cs-MBZ@ZnO half dose of MBZ starting from the 3rd dpi for three successive days for the treatment of the intestinal phase of infection. Results of the adult worm counts in all treated groups showed the highest percent (96.4%) reduction in G5 and G6. Improvement of the intestinal histopathological changes and the lowest intensity of vascular endothelial growth factor (VEGF) expression in the intestinal tissue were observed in groups G5 and G6 compared to the control group. The conclusion of biochemical results showed a significant increase in levels of aspartate transferase (AST), alanine transaminase (ALT), and globulin in the infected control group and a decrease in all treated groups. We recommended using the modified drug MBZ as an alternative drug to MBZ for trichinellid treatment, but the results need to be applied clinically.

Regulation of the HMGA2-SNAI2/CXCR4 axis in atherosclerosis and retinal neovascularization: new therapeutic insights.

Atherosclerosis (AS) is a vascular disease associated with endothelial damage, plaque formation, and retinal neovascularization (RNV), leading to visual impairment. Research indicates that vascular endothelial dysfunction, lipid deposition, and inflammatory responses contribute to the formation of plaques and atherosclerotic lesions. Among the common complications, studies have shown that RNV and the molecular mechanisms of AS hold significant clinical importance. In this study, we identified the overexpression of the gene heat shock protein 90 (HSP90) through transcriptome sequencing. Subsequent protein expression analysis and inhibition experiments in corresponding animal models confirmed the crucial role of HSP90 in the modulation of this disease. Research findings revealed an increase in the expression of HSP90, HMGA2, Snail family transcriptional repressor 2 gene (SNAI2), CXC chemokine receptor 4 (CXCR4), and vascular endothelial growth factor (VEGF) in atherosclerotic mouse tissues. Inhibition of HSP90 expression reduced vascular neovascularization and downregulated the expression of HMGA2 and VEGF. Given that HSP90 can promote HMGA2 expression, which, in turn, facilitates angiogenesis, we conducted lentiviral infection experiments on primary retinal endothelial cells obtained from atherosclerotic mice, confirming the regulatory role of HSP90 in modulating HMGA2 expression through the SNAI2/CXCR4 signaling pathway and its involvement in retinal endothelial neovascularization. In conclusion, our study highlights the significant regulatory role of HSP90 in AS-induced RNV, providing a new target for disease treatment. Furthermore, this research extensively explores the mechanism of HSP90 in regulating RNV and associated signaling pathways, offering novel insights and laying a solid foundation for future studies in this disease domain.

Exosomal microRNA-21-5p from gastric cancer cells promotes angiogenesis by targeting LEMD3 in human endothelial cells

Abstract Objectives The effect of exosome-derived miR-21-5p from gastric cancer (GC) on angiogenesis remains unclear. This study aims to examine the angiogenic impact of GC exosome-derived miR-21-5p. Methods Exosomes were isolated from GC cells and co-cultured with human umbilical vein endothelial cells (HUVECs). miR-21-5p levels in HUVECs were measured by qPCR. Flow cytometry was used to assess apoptosis, and the Cell Counting Kit-8 was used to assess cell growth. Bioinformatics analysis was used to identify the miR-21-5p target genes, which were then verified by dual-luciferase gene reporter experiments. qPCR and western blotting were employed to assess the expression of genes and proteins, respectively. Functional rescue assays were conducted to verify that miR-21-5p regulates endothelial cell function by targeting the LEM domain containing 3 (LEMD3). Additionally, cell migration was assessed using a scratch assay. Results Co-cultivation with GC-derived exosomes improved cell survival, decreased apoptosis, and raised miR-21-5p levels in HUVECs. Increases in vascular endothelial growth factor A (VEGFA) and the TGF-β/Smad signaling pathway were seen. It was shown that miR-21-5p targets LEMD3. The biological effects of miR-21-5p were lessened when miR-21-5p was inhibited, or LEMD3 was overexpressed. Conclusions By targeting LEMD3, miR-21-5p in GC cell exosomes stimulates angiogenesis by triggering the TGF-β/Smad signaling cascade and upregulating VEGFA expression. This leads to increased proliferation, survival, and migration of HUVECs, underscoring the potential of targeting this pathway in cancer therapy.

Effect of photobiomodulation and corticopuncture methods on tooth displacement and gene expression: animal study.

The aim of this study was to evaluate the expression levels of vascular endothelial growth factor (VEGF), Peroxiredoxin 1 (PRX1), glucose transporter 1 (GLUT1) and type I collagen (COL1) and the rate of tooth movement comparing 3 accelerated tooth movement (ATM) methods: Corticopuncture (CP), photobiomodulation (PBM) and the combined technique (CP + PBM) on days 1, 3, 7 and 14. Orthodontic tooth movement was induced in 24 male Wistar rats. CP procedure included three perforations: two in the palate and one mesial to the molars. GaAlAs diode laser irradiation was performed on days 0, 2, 4 and 6, totaling 4 irradiations. 14 days (810nm, 100 mW, 15s). Gingival tissue was collected from the cervical area of both first molars and qPCR was performed to isolate and quantify mRNA levels. All ATM groups showed increased tooth displacement compared to control after 14 days (20% for PBM; 40% for CP and 60% for CP + PBM). PBM showed higher VEGF expression on days 1,3 and 7 followed by CP and CP + PBM. PRX1 levels increased on days 1 and 3 in PBM and CP + PBM. GLUT1 increased on day 3 in all groups. No difference was found on levels of VEGF, PRX1 and GLUT1 among the groups on day 14, except for COL1 which increased significantly in PBM group. All ATM methods showed higher expression of all of VEGF, PRX1, GLUT1, COL1 than control group. PBM and CP + PBM groups had more expression related to angiogenesis, glucose uptake, oxidative stress and collagen synthesis.

The Potency of Centella asiatica Leaf Extract on VEGF Expression and Angiogenesis in Second-Degree Burn Wound in Mice

Burn injuries present a significant global health challenge, with the highest incidence rates reported in Southeast Asia, including Indonesia. Healing burn wounds is a complex and dynamic process involving various cellular and molecular mechanisms, prominently featuring the role of Vascular Endothelial Growth Factor (VEGF) in tissue regeneration and wound repair. VEGF is crucial for inducing and regulating angiogenesis and supplying oxygen and nutrients to the healing tissue. This study aims to evaluate the potential of pegagan (Centella asiatica) leaf extract cream 1%, 3%, and 5% daily for 14 days in enhancing VEGF expression and angiogenesis in second-degree burn wounds in mice (Mus musculus). This study investigates the application of C. asiatica extract cream on second-degree burn wounds in mice, comparing its effects on VEGF protein expression and angiogenesis to those of base cream and silver sulfadiazine cream, with outcomes evaluated using immunohistochemistry (IHC) and hematoxylin and eosin (HE) staining methods. Our findings suggest that C. asiatica extract cream promotes reduced burn wound size, significant upregulated VEGF expression, and enhanced angiogenesis in treating burn wounds compared to positive control, with a 5% dose having the best result. The study concludes that C. asiatica extract cream may effectively treat burn wound healing through enhancing VEGF expression and angiogenesis.

Gene Expression Modulation of Markers Involved in Bone Formation and Resorption by Bisphenol A, Bisphenol F, Bisphenol S, and Bisphenol AF.

Bisphenol A (BPA) and its analogs (BPF, BPS, and BPAF) are recognized for inducing detrimental effects on various tissues, including bone. The aim of this study is to investigate their impact on information and repair processes, specifically focusing on vascular endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1), and the receptors for transforming growth factor β (TGFR1, TGFR2, and TGFR3). Human osteoblasts isolated through primary culture from bone samples of healthy volunteers were subjected to cultivation in the presence of various dosage levels (10-5, 10-6, or 10-7 M) of BPA, BPF, BPS, or BPAF for 24 h. Gene expressions of RANKL, OPG, TGF-β1, TGFR1, TGFR2, TGFR3, and VEGF were analyzed by real-time polymerase chain reaction (RT-PCR). All experiments included untreated cells as controls. Expressions of RANKL and OPG were dose-dependently downregulated by the presence of all tested bisphenols (BPs) except for BPAF, whose presence upregulated OPG expression at all three doses. TGF-β1 expression was downregulated by all BP treatments, and TGF-β1 receptor expression was also downregulated as a function of the BP and dose. VEGF expression was downregulated in the presence of BPF and BPAF at all three doses and in the presence of BPA at the two higher doses (10-5, and 10-6 M), but it was not changed by the presence of BPS at any dose. The inhibition of both RANKL and OPG by the BPs, with a higher %inhibition of RANKL than of OPG, appears to rule out BP-induced activation of osteoclastogenesis via RANKL/RANK/OPG. Nevertheless, the effect of the BPs on the expression by osteoblasts of TGF-β1, TGF-β receptors, and VEGF indicates that these compounds can be responsible for major molecular changes in this cell population, contributing to their adverse effects on bone tissue.

Application of extracorporeal shockwave to regulate subchondral bone homeostasis through tumor necrosis factor-α/hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway in treatment of talus bone marrow edema.

To investigate the effect of extracorporeal shock wave on the treatment of talus bone marrow edema by regulating subchondral bone homeostasis through tumor necrosis factor-α (TNF-α)/hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF) signaling pathway. A total of 81 patients with talus bone marrow edema admitted to our hospital from May 2019 to May 2021 were studied and divided into control group (n = 40) and extracorporeal shock group (n = 41) according to random number table method. The control group was given conventional treatment, and the extracorporeal shock group was combined with extracorporeal shock wave therapy on the basis of the control group. The expression of TNF-α, HIF-1α, and VEGF in the 2 groups were compared, pain degree, and the area of talus bone marrow edema was evaluated by magnetic resonance imaging. The visual analogue scale scores of 1 month, 2 months and 5 months after treatment were decreased in both groups, and the extracorporeal shock group was lower than the control group (P < .05). After 5 months of treatment, the expressions of TNF-α and HIF-1α were decreased in both groups, and the extracorporeal shock group was lower than the control group, VEGF was increased, and the extracorporeal shock group was higher than the control group (P < .05), and the western blot expression levels of TNF-α, HIF-1α and VEGF in the extracorporeal shock group were higher than the control group (P < .05). The dorsiflexion motion and plantar flexion motion of both groups were increased, and the extracorporeal shock group was higher than the control group (P < .05). Extracorporeal shock wave therapy can regulate subchondral bone homeostasis through TNF-α/HIF-1α/VEGF signaling pathway to treat talus bone marrow edema, reduce the pain degree of talus bone marrow edema, and improve ankle joint function.

VEGF inhibition increases expression of HIF-regulated angiogenic genes by the RPE limiting the response of wet AMD eyes to aflibercept

Neovascular age-related macular degeneration (nvAMD) is the leading cause of severe vision loss in the elderly in the developed world. While the introduction of therapies targeting vascular endothelial growth factor (VEGF) has provided the first opportunity to significantly improve vision in patients with nvAMD, many patients respond inadequately to current anti-VEGF therapies. It was recently demonstrated that expression of a second angiogenic mediator, angiopoietin-like 4 (ANGPTL4), synergizes with VEGF to promote choroidal neovascularization (CNV) in mice and correlates with reduced response to anti-VEGF therapy in patients with nvAMD. Here, we report that expression of ANGPTL4 in patients with nvAMD increases following treatment with anti-VEGF therapy and that this increase is dependent on accumulation of hypoxia-inducible factor (HIF)-1α in response to inhibition of VEGF/KDR signaling in the retinal pigment epithelium (RPE). We therefore explored HIF-1 inhibition with 32-134D, a recently developed pharmacologic HIF-inhibitor, for the treatment of nvAMD. 32-134D prevented the expression of both VEGF and ANGPTL4 and was at least as effective as aflibercept in treating CNV in mice. Moreover, by preventing the increase in HIF-1α accumulation in the RPE in response to anti-VEGF therapy, combining 32-134D with aflibercept was more effective than either drug alone for the treatment of CNV. Collectively, these results help explain why many patients with nvAMD respond inadequately to anti-VEGF therapy and suggest that the HIF inhibitor 32-134D will be an effective drug-alone or in combination with current anti-VEGF therapies-for the treatment of patients with this blinding disease.

Wound healing after surgical therapy for multiple myeloma: a case-control study.

The aim of this study was to observe the surgical wound healing process in patients with multiple myeloma who had undergone surgery. We collected clinical data on patients with multiple myeloma and observed wound healing following surgical therapy. Additionally, we compared the expression of angiogenesis markers in patients with and without multiple myeloma (undergoing surgical excision of other tumour tissues). In patients who had multiple myeloma bone disease, we examined several clinical features: haemoglobin levels; albumin levels; blood glucose levels; and surgery programme. We then compared expression levels of the angiogenesis markers CD31, CD34 and vascular endothelial growth factor (VEGF) in samples scraped from the skin margin of the surgical incision in 12 patients without multiple myeloma (control) and nine patients with multiple myeloma. All 61 patients with multiple myeloma observed showed no disunion, no delayed union and no infection in their wound healing. CD31 and VEGF expression was higher in the nine patients with multiple myeloma compared with the 12 control patients without. We observed no difference in CD34 expression between control and experimental groups. The results of this study suggest that patients with multiple myeloma who have undergone surgery recover well and produce higher quantities of new vessels compared with patients without multiple myeloma. This occurs through increased expression of CD31 and VEGF, angiogenic factors which promote wound healing. We did not observe higher expression of these factors contributing to increased incisional implantation metastasis.

Intravenous Regeneration-associated Cell Transplantation Enhances Tissue Recovery in Mice with Acute Ischemic Stroke.

Previously, we reported that transplantation of regeneration-associated cells (RACs) via the ipsilateral external carotid artery reduced stroke volume in mice with permanent occlusion of the middle cerebral artery (MCA). However, intracarotid arterial transplantation is invasive and requires skill, and severe complications may occur, such as thromboembolism, infection, and decreased cerebral blood flow. This study aimed to investigate the efficacy of intravenous injection of RACs in reducing stroke volume and increasing anti-inflammatory and angiogenic factors in mice with focal cerebral ischemia. Mice with occluded MCAs received intravenous injections of phosphate-buffered saline (PBS) (control), low-dose RACs, or high-dose RACs. The proximal part of the left MCA was occluded to induce permanent focal ischemia. After 3 days, we administered PBS or low-dose (1 × 104 /50 µL) or high-dose RACs (1 × 105 /50 µL) through the tail vein and assessed the infarct volume on day 7. High-dose RACs significantly decreased infarct volume compared to PBS, whereas low-dose RACs showed no effect. The number of interleukin-10 (IL-10)-positive and vascular endothelial growth factor (VEGF)-positive cells in the peri-infarct area on day 7 was significantly higher in mice treated with low-dose and high-dose RACs than in the PBS control group. Intravenous injection of RACs can reduce ischemic stroke volume; however, a higher dose of RACs is required than the dose used in intraarterial transplantation. By assessing IL-10 and VEGF expression, the study sheds light on the underlying mechanisms of RAC therapy, revealing its potential anti-inflammatory and angiogenic properties in the treatment of cerebral ischemia.

Molecular Imaging of Melanoma VEGF-expressing Tumors through [99mTc]Tc-HYNIC-Fab(Bevacizumab).

Angiogenesis is a process that many tumors depend on for growth, development, and metastasis. Vascular endothelial growth factor (VEGF) is one of the major players in tumor angiogenesis in several tumor types, including melanoma. VEGF inhibition is achieved by bevacizumab, a humanized monoclonal antibody that binds with high affinity to VEGF and prevents its function. In order to successfully enable in vivo VEGF expression imaging in a murine melanoma model, we previously labeled bevacizumab with [99mTc]Tc. We observed that this was feasible, but it had prolonged blood circulation and delayed tumor uptake. The aim of this study was to develop a radiolabeled Fab bevacizumab fragment, [99mTc]Tc-HYNICFab( bevacizumab), for non-invasive in vivo VEGF expression molecular imaging. Flow cytometry was used to examine VEGF presence in the murine melanoma cell line (B16-F10). Bevacizumab was digested with papain for six hours at 37°C to produce Fab(bevacizumab), which was then conjugated to NHS-HYNIC-Tfa for radiolabeling with [99mTc]Tc. Stability and binding affinity assays were also evaluated. Biodistribution and single photon emission computed tomography/computed tomography (SPECT/CT) were performed at 1, 3, and 6 h (n = 4) after injection of [99mTc]Tc-HYNIC-Fab(Bevacizumab) in normal and B16-F10 tumor-bearing C57Bl/6J mice. Using flow cytometry, it was shown that the B16-F10 murine melanoma cell line has intracellular VEGF expression. Papain incubation resulted in the complete digestion of bevacizumab with good purity and homogeneity. The radiolabeling yield of [99mTc]Tc-HYNIC-Fab(bevacizumab) was 85.00 ± 6.06%, with a specific activity of 291.87 ± 18.84 MBq/mg (n=3), showing in vitro stability. Binding assays demonstrated significant intracellular in vitro VEGF expression. Fast blood clearance and high kidney and tumor uptake were observed in biodistribution and SPECT/CT studies. We present the development and evaluation of [99mTc]Tc-HYNIC-Fab(bevacizumab), a novel molecular VEGF expression imaging agent that may be used for precision medicine in melanoma and potentially in other VEGF-expressing tumors.

Updated Survival Follow-Up for Phase Ib Trial of the Histone Deacetylase Inhibitor Abexinostat With Pazopanib in Patients With Solid Tumor Malignancies.

Histone deacetylase (HDAC) inhibition downregulates hypoxia-inducible factor-1α and modulates multiple metabolomic pathways relevant in cancer. Here we report a potential novel biomarker to predict exceptional responders (>3 years) in patients receiving HDAC and vascular endothelial growth factor (VEGF) inhibition. Patients with solid tumor malignancies were enrolled in this phase Ib trial of abexinostat (4/7 ×21 days) and pazopanib (28/28 days), with a dose expansion in renal cell carcinoma (RCC). Plasma was analyzed for metabolomics and peripheral blood mononuclear cells (PBMCs) for VEGF and HDAC2 expression levels. Fifty-one patients were enrolled: n = 36 patients in dose escalation and n = 15 in dose expansion. After the initial report in 2017, six patients had remained on study: four with RCC and one each with medullary thyroid and thymic neuroendocrine carcinoma. One patient with RCC remains on treatment for >11 years after progression on five systemic therapies. Overall, the median duration of therapy measured 5.6 (1-133) months. The median duration of therapy in exceptional responders measured 44.1 (39.8-133+) months. The median overall survival in patients with high PBMC HDAC2 expression versus low HDAC2 was 32.3 versus 9.2 months (P = .004) for all patients and 43.3 versus 25.1 months for patients with RCC (P = .09). Exceptional responders had lower kynurenine levels both pre- and post-treatment (P = .002, P < .001, respectively). HDAC2 and kynurenine expression levels were inversely correlated (P = .02). Abexinostat added to pazopanib shows extended tolerability and long-term responses and survival. PBMC HDAC2 levels, the abexinostat target, are relevant predictors of response. In addition, metabolomic assessment points to kynurenine as a predictor for exceptional response to combined VEGF plus HDAC inhibition.

Uniaxial static strain enhances osteogenic and angiogenic potential under hypoxic conditions in distraction osteogenesis

ObjectiveBone incision leads to interrupted and sluggish blood flow in the process of distraction osteogenesis (DO), creating a hypoxia (0–2% oxygen tension) at the center of the bone callus. This hypoxia is critical in the coupling of osteogenesis and angiogenesis during DO. This study aimed to investigate the effect of Uniaxial Static Strain (USS) on osteogenesis in osteoblasts under hypoxic conditions, with a focus on the expression of osteogenic markers and angiogenic factors.MethodsThe USS was made by a multi-unit tension compression device.Osteoblasts were subjected to 10% USS made under hypoxic conditions to mimic the process of DO in vitro. The cell proliferation, alkaline phosphatase (ALP) activity, mineralized nodule formation, and expression of osteogenic and angiogenic markers were evaluated by using a CCK-8 assay, alkaline phosphatase (ALP) staining, ALP activity assay, alizarin red S staining, qRT-PCR, Western blotting and ELISA.ResultsHypoxia inhibited osteoblast cell proliferation, ALP activity, mineralized nodule formation, and the expression of runt-related transcription factor 2 (Runx- 2), osteopontin(OPN), osteocalcin (OCN), collagen type I (Col1a1). Conversely, hypoxia upregulated the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF), which are associated with angiogenesis. However, the application of USS enhanced osteoblasts’ osteogenic capacity and upregulated angiogenic factors under hypoxic conditions.ConclusionUSS can enhance osteogenesis in osteoblasts under hypoxic conditions. Moreover, it may stimulate angiogenesis by promoting the expression of VEGF, which further contributes to bone formation. This finding provides important implications for understanding the mechanisms involved in bone regeneration and may have clinical applications in optimizing the effectiveness of DO techniques.

New insights in the mechanisms of opioid analgesia and tolerance: Ultramicronized palmitoylethanolamide down-modulates vascular endothelial growth factor-A in the nervous system

Growing evidence suggests that opioid analgesics modulate angiogenesis during pathophysiological processes. Vascular endothelial growth factor-A (VEGF-A) was recently proposed to be involved in pain development. To date, no anti-angiogenic drug is used for pain management. When administered in a bioavailable formulation, (i.e., ultramicronized) N-palmitoylethanolamine (PEA) delays the onset of morphine tolerance, improves morphine analgesic activity and reduces angiogenesis in in vivo models. This study aimed at investigating whether VEGF-A is involved in PEA-induced delay of morphine tolerance. The anti-VEGF-A monoclonal antibody bevacizumab was used as a reference drug. Preemptive and concomitant treatment with ultramicronized PEA delayed morphine tolerance and potentiated the analgesic effect of morphine, while counteracting morphine-induced increase of VEGF-A in the nervous system. Similar results were obtained when bevacizumab was administered together with morphine. Of note, bevacizumab showed an analgesic effect per se. In equianalgesic treatment regimens (obtained through increasing morphine doses and associating PEA), PEA resulted in lower expression of VEGF-A in dorsal root ganglia (DRG) and spinal cord compared to morphine alone. Similar results were observed for plasma levels of the soluble VEGF receptor 1 (sFLT-1). Moreover, in morphine-treated animals, two pain-related genes (i.e., Serpina3n and Eaat2) showed a more than 3-fold increase in their expression at spinal cord and DRG level, with the increase being significantly counteracted by PEA treatment. This study supports the hypothesis that the effects of PEA on morphine analgesia and tolerance may be mediated by the down-modulation of VEGF-A and sFLT-1 in the nervous system and plasma, respectively.

Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor A (VEGF-A) expressions in Ethiopian female breast cancer and their association with histopathologic features.

Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor (VEGF) play important role in breast tumor growth, invasion, metastasis, patient survival and drug resistance. The aim of this study was to evaluate the protein expression status of EGFR and VEGF-A, as well as their association with hormone receptor status and histopathological characteristics in the invasive type of female breast cancer among Ethiopians. The primary breast tumor tissues were obtained from 85 Ethiopian invasive breast cancer cases that underwent modified radical mastectomy (MRM) from June 2014 to June 2015. Their FFPE blocks were analyzed for EGFR and VEGF protein expressions using immunohistochemical techniques. The expressions were also correlated with histopathologic features. Epidermal growth factor receptor over-expression was observed in 22% of the tumor samples. VEGF-A expression was negative in 13.41%, low in 63.41%, moderate in 20.73%, and high in 2.44%. EGFR expression, but not VEGF-A, showed a significant inverse correlation with both estrogen receptor (ER) (P = 0.01) and progesterone receptor (PR) statuses (P = 0.04). EGFR and VEGF expressions did not show significant association with tumor size, grade, lymph node status or age at diagnosis. Epidermal growth factor receptor expression was most likely associated with ER and PR negative tumors. Assessments of multiple molecular markers aid to understand the biological behavior of the disease in Ethiopian population. It might also help to predict which group of patients might get more benefit from the selected treatment strategies and which are not.

Effects of propranolol on the biological behavior of human umbilical vein endothelial cells and the expression of SOX18, MMP-7, and VEGFA

To investigate the effects of propranolol on the proliferation, apoptosis, migration, and tube formation ability of human umbilical vein endothelial cells (HUVEC), as well as its impact on the expression of sex-determining region Y-box 18 (SOX18), matrix metalloproteinase-7 (MMP-7), and vascular endothelial growth factor A (VEGFA). HUVEC were treated with different concentrations of propranolol, and cell viability was assessed using the CCK-8 method to determine the optimal concentration and treatment duration. The experiment consisted of a control group and groups treated with different concentrations of propranolol (50, 100, 150 μmol/L). Apoptosis, migration, and tube formation of HUVEC were observed using flow cytometry, wound healing assays, and tube formation assays. Western blot and real-time quantitative PCR were used to detect the expression levels of SOX18, MMP-7, and VEGFA proteins and mRNA. Compared to the control group, the apoptosis rate in the propranolol treatment groups increased significantly (P<0.05), and it rose significantly with increasing drug concentration (P<0.05). The wound healing rate decreased in the propranolol treatment groups, and both the number of tube formation nodes and total tube length were reduced (P<0.05). The expression levels of SOX18, MMP-7, and VEGFA proteins and mRNA were downregulated in the propranolol treatment groups (P<0.05). Propranolol can inhibit the proliferation, migration, and tube formation ability of HUVEC and promote cell apoptosis, resulting in decreased expression levels of SOX18, MMP-7, and VEGFA.