Vascular Endothelial Growth factor-C Research Articles

HHEX is a transcriptional regulator of the VEGFC/FLT4/PROX1 signaling axis during vascular development

Formation of the lymphatic system requires the coordinated expression of several key regulators: vascular endothelial growth factor C (VEGFC), its receptor FLT4, and a key transcriptional effector, PROX1. Yet, how expression of these signaling components is regulated remains poorly understood. Here, using a combination of genetic and molecular approaches, we identify the transcription factor hematopoietically expressed homeobox (HHEX) as an upstream regulator of VEGFC, FLT4, and PROX1 during angiogenic sprouting and lymphatic formation in vertebrates. By analyzing zebrafish mutants, we found that hhex is necessary for sprouting angiogenesis from the posterior cardinal vein, a process required for lymphangiogenesis. Furthermore, studies of mammalian HHEX using tissue-specific genetic deletions in mouse and knockdowns in cultured human endothelial cells reveal its highly conserved function during vascular and lymphatic development. Our findings that HHEX is essential for the regulation of the VEGFC/FLT4/PROX1 axis provide insights into the molecular regulation of lymphangiogenesis.

Abstract 1750: Inducing therapeutic lymphangiogenesis for potentiating cancer immunotherapy in melanoma

Abstract Tumor-associated lymphangiogenesis is well known to promote metastasis and to correlate with poor prognosis in melanoma and other cancers, yet its functional roles in antitumor immunity had remained largely unexplored. In a recent study, we investigated the role of the vascular endothelial growth factor C (VEGF-C)—the main driver of lymphangiogenesis—in regulating antitumor immune response. We showed that in the B16 F10 injectable melanoma model, VEGF-C inhibition prevented T cells from infiltrating the tumor and reduced the efficacy of immunotherapies. Here, we sought to extend this concept to exploit the immunologic benefits of VEGF-C for therapeutic applications. We engineered a VEGF-C variant with an extracellular matrix-binding domain (MB-VEGF-C) allowing retention into the microenvironment after intratumoral injection. MB-VEGF-C induced increased lymphatic vasculature in tumors without reducing tumor growth rate. In an autochthonous melanoma model with reduced immune infiltrates, MB-VEGF-C intratumoral injections promoted the infiltration of cross-presenting DCs, as well as CD8+ and CD4+ T cells. Crucially, in these tumors that are normally unresponsive to immune checkpoint blockade (ICB), we could restore the responsiveness to ICB by delivering MB-VEGF-C together with immune adjuvants. Taken together, our findings suggest that VEGF-C and tumor-associated lymphangiogenesis, albeit promoting metastasis, can also be exploited to promote a T cell-inflamed microenvironment and to subsequently potentiate immunotherapies. Citation Format: Lambert Potin, Lea Maillat, Priscilla S. Briquez, Jeffrey A. Hubbell, Melody A. Swartz. Inducing therapeutic lymphangiogenesis for potentiating cancer immunotherapy in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1750.

High MUC2 Mucin Biosynthesis in Goblet Cells Impedes Restitution and Wound Healing by Elevating Endoplasmic Reticulum Stress and Altered Production of Growth Factors

Intestinal epithelial cell wound healing involves cell migration, proliferation, and differentiation. Although numerous studies have analyzed the migration of absorptive epithelial cells during wound healing, it remains unclear how goblet cells restitute and how MUC2 mucin production affects this process. In this study, we examined the role of high MUC2 production in goblet cell migration during wound healing and demonstrated that during high MUC2 output, goblet cells migrated slower because of impaired production of wound healing factors and endoplasmic reticulum (ER) stress. Two goblet cell lines, HT29-H and HT29-L, that produced high and low MUC2 mucin, respectively, were used. HT29-L healed wounds faster than HT29-H cells by producing significantly higher amounts of fibroblast growth factor (FGF) 1, FGF2, vascular endothelial growth factor-C, and matrix metallopeptidase 1. Predictably, treatment of HT29-H cells with recombinant FGF2 significantly enhanced migration and wound healing. High MUC2 biosynthesis in HT29-H cells induced ER stress and delayed migration that was abrogated by inhibiting ER stress with tauroursodeoxycholic acid and IL-22. FGF2- and IL-22-induced wound repair was dependent on STAT1 and STAT3 signaling. During wound healing after dextran sulfate sodium-induced colitis, restitution of Math1M1GFP+ goblet cells occurred earlier in the proximal colon, followed by the middle and then distal colon, where ulceration was severe. We conclude that high MUC2 output during colitis impairs goblet cell migration and wound healing by reducing production of growth factorscritical in wound repair.

High serum vascular endothelial growth factor C predicts better relapse-free survival in early clinically node-negative breast cancer.

A recent meta-analysis indicated that higher tumoral expression of vascular endothelial growth factor C (VEGF-C) was related to poorer relapse-free and overall survival in breast cancer patients. However, a retrospective study found that higher circulating VEGF-C levels were associated with better survival in breast cancer patients. In 2009, we initiated a prospective study to determine the utility of preoperative serum VEGF-C levels for predicting the risk of sentinel lymph node involvement in early breast cancer and to assess serum VEGF-C levels as a prognostic factor for relapse-free and overall survival. We analyzed serum samples from 174 patients with early breast cancer who underwent sentinel lymph node biopsies. VEGF-C levels were determined using an ELISA. Serum VEGF-C levels were normally distributed, with a median value of 6561.5 pg/mL, and did not correlate with any other clinical or pathological variables. During a median follow-up period of 58 months, the five-year relapse-free survival rate was higher in patients with VEGF-C levels above the median than in patients with lower levels (95.3% vs. 85.9%, p < 0.04). No association was found between VEGF-C levels and overall survival. Our study demonstrates that the prognosis was better for early breast cancer patients with high serum VEGF-C levels.

Steroid receptor coactivator-1 interacts with NF-κB to increase VEGFC levels in human thyroid cancer.

Thyroid cancer is the most common endocrine cancer, and has a high incidence of lymphatic metastasis. Vascular endothelial growth factor C (VEGFC) is essential for development of lymphatic vessels and lymphatic metastases during carcinogenesis. Steroid receptor coactivator-1 (SRC-1) interacts with nuclear receptors and transcription factors to promote tumor proliferation and metastasis. However, the correlation between SRC-1 and VEGFC levels in the lymphatic metastases of thyroid cancer remains unclear. We analyzed 20-paired specimens of thyroid cancer tissue and normal thyroid tissue and found increased levels of SRC-1 and VEGFC proteins in 13/20 and 15/20 thyroid cancer specimens, respectively, when compared with those levels in specimens of normal thyroid tissue. A high level of SRC-1 expression was positively correlated with VEGFC and lymphatic endothelial cell marker LYVE-1 expression. Papillary thyroid carcinoma cell line TPC-1 displayed high levels of SRC-1 and VEGFC expression and was selected for stable knockdown of SRC-1 in vitro. Inhibition of SRC-1 significantly reduced the VEGFC levels in TPC-1 cells. We found that SRC-1 binds to transcription factor NF-kB (p50/p65), and that this coactivation complex directly promoted VEGFC transcription, which could be abrogated by SRC-1 knockdown. Up-regulated NF-kB signaling was also confirmed in thyroid cancer tissues. In vivo studies showed that SRC-1 knockdown restricted tumor growth, reduced the numbers of LYVE-1-positive lymphatic vessels, and decreased the levels of VEGFC in tumor tissues. These results suggest a tumorigenic role for SRC-1 in thyroid cancer via its ability to regulate VEGFC expression.

Expression of 4E-BP1 and phospho-4E-BP1 correlates with the prognosis of patients with clear cell renal carcinoma.

BackgroundEukaryotic translation initiation factor 4E (eIF4E) is a key regulator of protein synthesis. Changes in eIF4E activity disproportionally affect the translation of a subset of oncogenic mRNAs in some cancers.Materials and methodsWe have assessed the expression levels of vascular endothelial growth factor C (VEGFC), eIF4E, eIF4E-binding proteins (4E-BPs) and phospho-4E-BP1 in clear cell renal carcinoma (ccRCC; n=101) using immunohistochemistry and analyzed the relevant mRNA levels and survival using online databases.ResultsThe protein levels of VEGFC, an eIF4E-regulated gene, were upregulated in ccRCC tissues compared with adjacent normal renal tissues, indicating an enhanced eIF4E activity in ccRCC. The expression of eIF4E had no significant changes in ccRCC tissues. However, 4E-BP1 and phospho-4E-BP1 were found to be overexpressed in ccRCC tissues (P<0.05), and the high mRNA and protein levels of 4E-BP1 and phospho-4E-BP1 correlated with an unfavorable clinical outcome in ccRCC patients. Meanwhile, the mRNA expression of PIK3CD and PIK3CG were enhanced in ccRCC.ConclusionFrom these results, we could infer that the increase in eIF4E activity may be caused by the increased phospho-4E-BP1 level, which was probably due to the activation of phosphoinositide 3-kinase (PI3K) pathway.

Impact of fenofibrate on choroidal neovascularization formation and VEGF-C plus VEGFR-3 in Brown Norway rats

ObjectiveThis study aims to explore the possible role of fenofibrate in inhibiting choroidal neovascularization (CNV) in Brown Norway (BN) rats. MethodsBN rats underwent binocular retinal laser photocoagulation to induce CNV. On day one, fenofibrate was injected into the vitreous cavity of rats in the control and experimental groups. Fundus fluorescein angiography (FFA), isolectin B4-FITC staining, immunofluorescence staining, qRT-PCR and western blot were performed at 1, 2, 3 and 4 weeks to observe the morphological changes of CNV and the expression of the vascular endothelial growth factor C (VEGF-C) and the vascular endothelial growth factor receptor-3 (VEGFR-3). ResultsCNV with the spontaneous gradual regression and scarring phenomenon appeared in BN rats. In neovascularization, VEGF-C was mainly distributed in the ganglion cell layer, while VEGFR-3 was mainly expressed in the choroid. In the control group, choroidal VEGF-C initially increased, and subsequently decreased, while VEGFR-3 level maintained a constant level after the decrease. Both had a decreasing expression in the retina. The early formation of CNV was significantly weakened in the experimental group, but there was no difference in the later period. VEGF-C and VEGFR-3 expression in the choroid and retina were lower than in the control group. Furthermore, VEGFR-3 protein was not expressed in the retina. However, this gradually increased in the early period and declined in the terminal stage in the choroid. ConclusionVEGF-C and VEGFR-3 participated in the laser-induced CNV formation in BN rats. Fenofibrate could inhibit CNV formation.

Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2

BackgroundTumor-associated macrophages (TAMs) play a critical role in modulating the tumor microenvironment and promote tumor metastases. Our studies have demonstrated that ginsenoside Rh2 (G-Rh2), a monomeric compound extracted from ginseng, is a promising anti-tumor agent in lung cancer cells. However, it remains unclear whetherG-Rh2 can modulate the differentiation of TAMs and its interaction with tumor microenvironment. In this study, we investigated how G-Rh2 regulates the phenotype of macrophages and affects the migration of non-small cell lung cancer (NSCLC) cells.MethodsMurine macrophage-like RAW264.7 cells and human THP-1 monocyte were differentiated into M1 and M2 subsets of macrophages with different cytokines combination, which were further identified by flow cytometry with specific biomarkers. M2 macrophages were sorted out to co-culture with NSCLC cell lines, A549 and H1299. Wound healing assay was performed to examine the cell migration. Expression levels of matrix metalloproteinases 2 and 9 (MMP-2, − 9) and vascular endothelial growth factor-C (VEGF-C) were measured by RT-qPCR and western blot, and the release of VEGF in the supernatant was measured by a VEGF ELISA kit. Finally, modulation of TAMs phenotype and VEGF expression by G-Rh2 was examined in vivo.ResultsWe demonstrated that M2 subset of macrophages alternatively differentiated from RAW264.7 or THP-1cells promote migration of NSCLC cells. Further examinations revealed that NSCLC significantly increased the release of VEGF to the media and elevated the expression levels of VEGF at mRNA and protein levels after being co-cultured with M2 macrophages. Similar alterations in MMP-2 and MMP-9 were observed in NSCLC after being co-cultured. Of note,G-Rh2 had a potential to effectively convert M2 phenotype to M1 subset of macrophages. Importantly, G-Rh2 had a preference to decrease the expression levels of VEGF, MMP2, and MMP9 in co-cultured lung cancer cells, over than those in lung cancer cells without co-culturing. Consistently, G-Rh2 reduced M2 macrophage marker CD206 and VEGF expression levels in vivo.ConclusionsAll of these results suggested that M2 subset macrophages drive lung cancer cells with more aggressive phenotypes. G-Rh2 has a potential to convert TAMs from M2 subset to M1 in the microenvironment and prevents lung cancer cell migration, suggesting the therapeutic effects of G-Rh2onlung cancer.

Historical overview of lymphangiogenesis.

The investigation of the lymphatic system has a very long and intriguing history, with several medical figures which brought important contributions. Work on the lymphatic system began in the 17th century, and by the beginning of the 19th century the anatomy of most of the lymphatic system had been described. This system is an essential component of the immune system, as well as vital to the maintenance of fluid homeostasis within the body. In this review article, I summarize the most important contributions to this field, until to the discovery of a specific lymphangiogenic factor, namely vascular endothelial growth factor-c (VEGF-C) (summarizing figure).

Resistin facilitates VEGF-C-associated lymphangiogenesis by inhibiting miR-186 in human chondrosarcoma cells

Chondrosarcoma is a common primary malignant tumor of the bone that can metastasize through the vascular system to other organs. A key step in the metastatic process, lymphangiogenesis, involves vascular endothelial growth factor-C (VEGF-C). However, the effects of lymphangiogenesis in chondrosarcoma metastasis remain to be clarified. Accumulating evidence shows that resistin, a cytokine secreted from adipocytes and monocytes, also promotes tumor pathogenesis. Notably, chondrosarcoma can easily metastasize. In this study, we demonstrate that resistin enhances VEGF-C expression and lymphatic endothelial cells (LECs)-associated lymphangiogenesis in human chondrosarcoma cells. We also show that resistin triggers VEGF-C-dependent lymphangiogenesis via the c-Src signaling pathway and down-regulating micro RNA (miR)-186. Overexpression of resistin in chondrosarcoma cells significantly enhanced VEGF-C production and LECs-associated lymphangiogenesis in vitro and tumor-related lymphangiogenesis in vivo. Resistin levels were positively correlated with VEGF-C-dependent lymphangiogenesis via the down-regulation of miR-186 expression in clinical samples from chondrosarcoma tissue. This study is the first to evaluate the mechanism underlying resistin-induced promotion of LECs-associated lymphangiogenesis via the upregulation of VEGF-C expression in human chondrosarcomas. We suggest that resistin may represent a molecular target in VEGF-C-associated tumor lymphangiogenesis in chondrosarcoma metastasis.

RIP1 regulates TNF-α-mediated lymphangiogenesis and lymphatic metastasis in gallbladder cancer by modulating the NF-κB-VEGF-C pathway.

BackgroundTumor necrosis factor alpha (TNF-α) enhances lymphangiogenesis in gallbladder carcinoma (GBC) via activation of nuclear factor (NF-κB)-dependent vascular endothelial growth factor-C (VEGF-C). Receptor-interacting protein 1 (RIP1) is a multifunctional protein in the TNF-α signaling pathway and is highly expressed in GBC. However, whether RIP1 participates in the signaling pathway of TNF-α-mediated VEGF-C expression that enhances lymphangiogenesis in GBC remains unclear.MethodsThe RIP1 protein levels in the GBC-SD and NOZ cells upon stimulation with increasing concentrations of TNF-α as indicated was examined using Western blot. Lentiviral RIP1 shRNA and siIκBα were constructed and transduced respectively them into NOZ and GBC-SD cells, and then PcDNA3.1-RIP1 vectors was transduced into siRIP1 cell lines to reverse RIP1 expression. The protein expression of RIP1, inhibitor of NF-κB alpha (IκBα), p-IκBα, TAK1, NF-κB essential modulator were examined through immunoblotting or immunoprecipitation. Moreover, VEGF-C mRNA levels were measured by quantitative real-time polymerase chain reaction, VEGF-C protein levels were measured by immunoblotting and enzyme-linked immunosorbent assay, and VEGF-C promoter and NF-κB activities were quantified using a dual luciferase reporter assay. The association of NF-κB with the VEGF-C promoter was analysed by chromatin immunoprecipitation assay. A three-dimensional coculture method and orthotopic transplantation nude mice model were used to evaluate lymphatic tube-forming and metastasis ability in GBC cells. The expression of RIP1 protein, TNF-α protein and lymphatic vessels in human GBC tissues was examined by immunohistochemistry, and the dependence between RIP1 protein with TNF-α protein and lymphatic vessel density was analysed.ResultsTNF-α dose- and time-dependently increased RIP1 protein expression in the GBC-SD and NOZ cells of GBC, and the strongest effect was observed with a concentration of 50 ng/ml. RIP1 is fundamental for TNF-α-mediated NF-κB activation in GBC cells and can regulate TNF-α-mediated VEGF-C expression at the protein and transcriptional levels through the NF-κB pathway. RIP1 can regulate TNF-α-mediated lymphatic tube formation and metastasis in GBC cells both in vitro and vivo. The average optical density of RIP1 was linearly related to that of TNF-α protein and the lymphatic vessel density in GBC tissues.ConclusionWe conclude that RIP1 regulates TNF-α-mediated lymphangiogenesis and lymph node metastasis in GBC by modulating the NF-κB-VEGF-C pathway.

Cytokines regulating lymphangiogenesis.

Lymphatic vessels are established by differentiation of lymphendothelial progenitors during embryogenesis. Lymphangiogenesis, the formation of new lymphatic vessels from pre-existing ones is rare in the healthy adult but takes place during pathological conditions such as inflammation, tissue repair and tumor growth. Conditions of dysfunctional lymphatics exist after surgical interventions or in certain genetic diseases. A key lymphangiogenic stimulator is vascular endothelial growth factor-C (VEGFC) acting on VEGF receptor-3 (VEGFR3) expressed on lymphendothelial cells. Other cytokines may act directly to regulate lymphangiogenesis positively or negatively, or indirectly by inducing expression of VEGFC. This review describes different known lymphangiogenic cytokines, their mechanism of action and role in lymphangiogenesis in health and disease.

Vascular endothelial growth factor C promotes cervical cancer cell invasiveness via regulation of microRNA-326/cortactin expression

Vascular endothelial growth factor C (VEGF-C) accelerates cervical cancer metastasis, while the detailed mechanism remains largely unknown. Recent evidence indicates that microRNA play a crucial role in controlling cancer cell invasiveness. In the present study, we investigated the role of miR-326 in VEGF-C-induced cervical cancer cell invasion. VEGF-C expression was higher and miR-326 was much lower in primary cervical cancer specimens than that in non-cancerous specimens, and a negative correlation between VEGF-C and miR-326 was found. On cervical carcinoma cell line SiHa cells, treatment with VEGF-C downregulated miR-326 level and increased cortactin protein expression. Transfection with miR-326 mimic reversed cortactin expression induced by VEGF-C, suggesting that VEGF-C increased cortactin via downregulation of miR-326. VEGF-C activated c-Src and c-Src inhibitor PP2 abolished VEGF-C effect on miR-326 and cortactin expression, implying that VEGF-C regulated miR-326/cortactin via c-Src signaling. VEGF-C promoted SiHa cell invasion index, which was largely inhibited by transfection with miR-326 antagonist or by siRNA against cortactin. In conclusion, our findings implied that VEGF-C reduced miR-326 expression and increased cortactin expression through c-Src signaling, leading to enhanced cervical cancer invasiveness. This may shed light on potential therapeutic strategies for cervical cancer therapy.

VEGFR-3 signaling is regulated by a G-protein activator, activator of G-protein signaling 8, in lymphatic endothelial cells

Vascular endothelial growth factor C (VEGFC) and its cognate receptor VEGFR-3 play a key role in lymphangiogenesis. We previously reported that an ischemia-inducible Gβγ signal regulator, activator of G-protein signaling 8 (AGS8), regulated the subcellular distribution of vascular endothelial growth factor receptor-2 (VEGFR-2) and influenced VEGFA-induced signaling in vascular endothelial cells. Here, we report that AGS8 regulates VEGFR-3, which is another subtype of the VEGF receptor family, and mediates VEGFC signaling in human dermal lymphatic endothelial cells (HDLECs). VEGFC stimulated the proliferation of HDLECs and tube formation by HDLECs, which were inhibited by knocking down AGS8 by small interfering RNA (siRNA). AGS8 siRNA inhibited VEGFC-mediated phosphorylation of VEGFR-3 and its downstream molecules, including ERK1/2 and AKT. Analysis of fluorescence-activated cell sorting and immunofluorescence staining demonstrated that AGS8 knockdown was associated with a reduction of VEGFR-3 at the cell surface. Endocytosis inhibitors did not rescue the decrease of cell-surface VEGFR-3, suggesting that AGS8 regulated the trafficking of VEGFR-3 to the plasma membrane. An immunoprecipitation assay indicated that VEGFR-3 formed a complex including AGS8 and Gβγ in cells. These data suggest the novel regulation of VEGFC-VEGFR-3 by AGS8 in HDLECs and a potential role for AGS8 in lymphangiogenesis.

The expression of anterior gradient-2 and vascular endothelial growth factor-C in prostate cancer and clinical data

The expression of anterior gradient-2 and vascular endothelial growth factor-C in prostate cancer and clinical data

PGF and VEGF‐C secreted by preosteoclasts induce migration and differentiation of Mesenchymal stem cells

Osteoblast‐mediated bone formation is coupled to osteoclast‐mediated bone resorption. Several studies suggested that the molecular mechanisms by which osteoblasts control osteoclastogenesis and bone degradation. But, the recruitment of mesenchymal stem cells (MSCs) and their differentiation to osteoblasts is poorly understood. Here we show that preosteoclasts secrete placental growth factor (PGF) and vascular endothelial growth factor C (VEGF‐C) to induce migration and osteogenesis of MSCs. Conditioned media from preosteoclasts increased migration ability of MSCs and alkaline phosphatase activity and mineralization in osteoblast cultures. Osteoclast precursors induced PGF and VEGF‐C expression in response to receptor activator of nuclear factor‐κB ligand (RANKL), an essential cytokine for osteoclast differentiation. In vitro experiments revealed that migration was stimulated by rhPGF and rhVEGF‐C in a dose‐dependent manner. Furthermore, these factors increased osteoblast differentiation. Vascular endothelial growth factor receptor (VEGFR) inhibitor treatment reduced these responses. Taken together, PGF and VEGF‐C secreted by preosteoclasts may enhance the recruitment of MSCs and further induce osteogenesis.Support or Funding InformationThis work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF‐ 2015R1D1A4A01020104).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Investigating the Role of Vascular Endothelial Growth Factor in Hematopoietic Stem Cell Driven Perivascular Niche Remodeling

Hematopoiesis is the process that produces the entirety of mature blood cells. A balance between the quiescence and proliferation of hematopoietic stem cells (HSC) is essential to maintain the blood supply. During development, HSCs expand exponentially in an intermediate niche before migration to the adult bone marrow. In zebrafish, the intermediate niche is the caudal hematopoietic tissue (CHT), a vascular plexus in the tail of the embryo, and in mice it is the fetal liver. Previous work by Tamplin et al. 2015 showed that arrival of an HSC triggers endothelial cells (EC) in the CHT niche to remodel and form a pocket around the HSC. Our goal is to uncover the function of the EC pocket and the mechanism of its formation. One candidate for this signaling cascade is vascular endothelial growth factor C (VEGF‐C) and its receptors VEGFR2 and VEGFR3. Within all hematopoietic lineages, VEGF‐C is highly expressed in HSC. VEGFR2/3 are expressed on sinusoidal endothelial cells of the CHT and fetal liver niches. Previous studies have shown a decrease in hematopoietic progenitor numbers when VEGFR3 is knocked out in mouse embryos (Hamada, 2000). Our preliminary data in zebrafish embryos have agreed with these findings. We have seen a decrease in hematopoietic stem and progenitor cells in vivo marked by the transgenic reporter cd41:gfp and in whole mount in situ with c‐myb probe after knocking down VEGFR3 alone as well as in combination with VEGFR2. To further confirm this phenotype, we administered pan VEGFR inhibitor, Semaxinib, and saw a similar decrease in cd41:gfp+ progenitors. We plan to further investigate this phenotype with a variety of small molecule inhibitors, genetic mutants, and synthetic oligo “morpholino” knockdown. Once we establish the mechanism for VEGF signaling in the remodeling of the perivascular EC niche, we plan to investigate the downstream targets. Uncovering the signals that govern the interaction between HSCs and their perivascular niche is clinically applicable in improving the efficiency of HSC transplants.Support or Funding InformationDepartment of Pharmacology, Lung and Vascular Biology Training Grant; NIH T32 HL027829This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Inhibitory effect of Sumu (Lignum Sappan) plus Fuzi (Radix Aconiti Lateralis Praeparata) on a lung carcinoma model

OBJECTIVETo investigate the inhibitory effect of Sumu (Lignum Sappan) plus Fuzi (Radix Aconiti Lateralis Praeparata) (SF) on the growth and metastasis of Lewis lung carcinoma. METHODSA lung carcinoma model was established by subcutaneously inoculating Lewis lung carcinoma cells into C57BL/6 mice. The mice were randomly divided into four groups of 13 mice (control, low-dose, moderate-dose, and high-dose), and gavaged once-daily for 21 consecutive days. The rates of tumor inhibition, metastasis, and metastasis inhibition were observed. The differential expressions of sP-selectin and vascular endothelial growth factor C (VEGFC) were compared between the treatment groups and the control group. RESULTSThe tumor weights differed significantly between the treatment groups and the control group (P < 0.05). Administration of SF in the moderate-dosage and low-dose groups significantly inhibited the expression of sP-selectin and VEGFC (both P < 0.05), suggesting anti-tumor activity. CONCLUSIONSF can inhibit the growth and metastasis of Lewis lung carcinoma.

Effect of High VEGF-C mRNA Expression on Achievement of Complete Remission in Adult Acute Myeloid Leukemia

Although vascular endothelial growth factor-C (VEGF-C) is known to be expressed in acute myeloid leukemia (AML) blasts, the relevance of VEGF-C in the clinical setting remains to be fully explored. We examined the effect of VEGF-C on achievement of complete remission (CR) in adult de novo AML and immune cell population profiles according to VEGF-C mRNA expression. In comparison of VEGF-C expression between the no-CR and CR groups, the CR group showed a trend toward higher levels of plasma VEGF-C (P = .088), whereas mRNA expression of VEGF-C was downregulated (P = .008). Next, patients with continuous data for VEGF-C were divided into two groups (low vs. high) by a ROC curve analysis. The low- versus high-level groups for plasma VEGF-C (RR of 0.20, P = .030), mRNA expression of VEGF-C (RR of 18.75, P = .003), and the ratio of plasma level to mRNA expression (RR of 0.05, P = .007) were potential predictors of CR on univariate analysis. After adjusting for potential clinical factors including genetic group, multivariate analyses revealed that high VEGF-C mRNA expression was an independent risk factor for failure of induction chemotherapy. Furthermore, patients with high VEGF-C mRNA expression had a lower frequency of NKT and CD8+ cells and showed a trend for a lower frequency of NK cells. These results suggest that interruption of VEGF-C signaling might be a potential therapeutic target for antileukemic treatment in AML patients.

Expression of CD44v3, erythropoietin and VEGF-C in gastric adenocarcinomas: correlations with clinicopathological features.

To investigate the expression of CD44v3 and vascular endothelial growth factor C (VEGF-C) in gastric adenocarcinoma and the correlation with erythropoietin (EPO) and clinicopathological features. The expression of CD44v3, VEGF-C and EPO was evaluated by immunohistochemical staining in 169 gastric adenocarcinomas. The correlations between these parameters and patients' clinicopathological features were analyzed statistically. CD44v3 and VEGF-C were positive in 50 (29.6%) and 82 (48.5%) patients, respectively. High CD44v3 expression was associated with poor cellular differentiation, extensive lymph node metastasis and advanced stage of gastric adenocarcinoma (P <0.05). High VEGF-C expression was significantly correlated with Lauren type, extensive lymph node metastasis and advanced stage of gastric adenocarcinoma (P <0.05). Univariate analysis of survival demonstrated that patients with a strong CD44v3 immunoreaction had a significantly worse overall survival compared with patients showing a weak CD44v3 immunoreaction (log-rank test: P = 0.0449). The mean survival time of patients with low CD44v3 expression was 30.6 months, which was much longer than the 21.6 months in patients with high expression. In addition, a strong association between immunohistochemical expression of CD44v3 and EPO was noted. Increased expression of CD44v3 and VEGF-C may play a significant role in the carcinogenesis and progression of gastric adenocarcinoma. High CD44v3 expression may be a predictor of poor prognosis in gastric adenocarcinoma patients. We infer that some mechanisms may exist in regulating the expression of CD44v3 and EPO.