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      1. Author :
        Foster, Greg A.; Headen, Devon M.; González-García, Cristina; Salmerón-Sánchez, Manuel; Shirwan, Haval; García, Andrés J.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2017
      5. Publication :
        Biomaterials
      6. Products :
      7. Volume :
        113
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        VEGF; Microfluidics; Biomaterials; Hydrogels; Protein delivery; IVIS; IVIS FLI in vivo
      12. Abstract :
        Degradable hydrogels to deliver bioactive proteins represent an emerging platform for promoting tissue repair and vascularization in various applications. However, implanting these biomaterials requires invasive surgery, which is associated with complications such as inflammation, scarring, and infection. To address these shortcomings, we applied microfluidics-based polymerization to engineer injectable poly(ethylene glycol) microgels of defined size and crosslinked with a protease degradable peptide to allow for triggered release of proteins. The release rate of proteins covalently tethered within the microgel network was tuned by modifying the ratio of degradable to non-degradable crosslinkers, and the released proteins retained full bioactivity. Microgels injected into the dorsum of mice were maintained in the subcutaneous space and degraded within 2 weeks in response to local proteases. Furthermore, controlled release of VEGF from degradable microgels promoted increased vascularization compared to empty microgels or bolus injection of VEGF. Collectively, this study motivates the use of microgels as a viable method for controlled protein delivery in regenerative medicine applications.
      13. URL :
        http://www.sciencedirect.com/science/article/pii/S0142961216305956
      14. Call Number :
        PKI @ catherine.lautenschlager @ 12588
      15. Serial :
        14094
      1. Author :
        Güç, Esra; Briquez, Priscilla S.; Foretay, Didier; Fankhauser, Manuel A.; Hubbell, Jeffrey A.; Kilarski, Witold W.; Swartz, Melody A.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2017
      5. Publication :
        Biomaterials
      6. Products :
      7. Volume :
        131
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        VEGF-C; Fibrin; Lymphatics; Regenerative medicine; IVIS; IVIS FLI in vivo
      12. Abstract :
        Lymphangiogenesis occurs in inflammation and wound healing, yet its functional roles in these processes are not fully understood. Consequently, clinically relevant strategies for therapeutic lymphangiogenesis remain underdeveloped, particularly using growth factors. To achieve controlled, local capillary lymphangiogenesis with protein engineering and determine its effects on fluid clearance, leukocyte trafficking, and wound healing, we developed a fibrin-binding variant of vascular endothelial growth factor C (FB-VEGF-C) that is slowly released upon demand from infiltrating cells. Using a novel wound healing model, we show that implanted fibrin containing FB-VEGF-C, but not free VEGF-C, could stimulate local lymphangiogenesis in a dose-dependent manner. Importantly, the effects of FB-VEGF-C were restricted to lymphatic capillaries, with no apparent changes to blood vessels and downstream collecting vessels. Leukocyte intravasation and trafficking to lymph nodes were increased in hyperplastic lymphatics, while fluid clearance was maintained at physiological levels. In diabetic wounds, FB-VEGF-C-induced lymphangiogenesis increased extracellular matrix deposition and granulation tissue thickening, indicators of improved wound healing. Together, these results indicate that FB-VEGF-C is a promising strategy for inducing lymphangiogenesis locally, and that such lymphangiogenesis can promote wound healing by enhancing leukocyte trafficking without affecting downstream lymphatic collecting vessels.
      13. URL :
        http://www.sciencedirect.com/science/article/pii/S0142961217301825 http://ac.els-cdn.com/S0142961217301825/1-s2.0-S0142961217301825-main.pdf?_tid=1ef4b8b0-6dc8-11e7-b45d-00000aab0f01&acdnat=1500609374_b267d8eb122c08a034c2801886586ed8
      14. Call Number :
        PKI @ catherine.lautenschlager @ 13585
      15. Serial :
        14066
      1. Author :
        Rusckowski, Mary; Wang, Yuzhen; Blankenberg, Francis G; Levashova, Zoia; Backer, Marina V; Backer, Joseph M
      2. Title :
        Targeted scVEGF/177Lu radiopharmaceutical inhibits growth of metastases and can be effectively combined with chemotherapy
      3. Type :
        Journal Article
      4. Year :
        2016
      5. Publication :
        EJNMMI Research
      6. Products :
      7. Volume :
        6
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        VEGF receptors,; 177Lu,; Targeted radiopharmaceutical,; Breast cancer,; Metastases; IVIS; IVIS BLI in vivo
      12. Abstract :
        Background: scVEGF/177Lu is a novel radiopharmaceutical targeted by recombinant single-chain (sc) derivative of vascular endothelial growth factor (VEGF) that binds to and is internalized by vascular endothelial growth factor receptors (VEGFR). scVEGF/177Lu potential as adjuvant and neoadjuvant anti-angiogenic therapy was assessed in metastatic and orthotopic mouse models of triple-negative breast cancer. Methods: Metastatic lesions in Balb/c mice were established by intracardiac injection of luciferase-expressing 4T1luc mouse breast carcinoma cells. Mice with metastatic lesions received single intravenous (i.v.) injection of well-tolerated dose of scVEGF/177Lu (7.4 MBq/mouse) at day 8 after 4T1luc cell injection. Primary orthotopic breast tumors in immunodeficient mice were established by injecting luciferase-expressing MDA231luc human breast carcinoma cells into mammary fat pad. Tumor-bearing mice were treated with single injections of scVEGF/177Lu (7.4 MBq/mouse, i.v), or liposomal doxorubicin (Doxil, 1 mg doxorubicin per kg, i.v.), or with a combination of Doxil and scVEGF/177Lu given at the same doses, but two hours apart. “Cold” scVEGF-targeting conjugate was included in controls and in Doxil alone group. The effects of treatments were defined by bioluminescent imaging (BLI), computed tomography (CT), computed microtomography (microCT), measurements of primary tumor growth, and immunohistochemical analysis. Results: In metastatic model, adjuvant treatment with scVEGF/177Lu decreased overall metastatic burden and improved survival. In orthotopic primary tumor model, a combination of Doxil and scVEGF/177Lu was more efficient in tumor growth inhibition than each treatment alone. scVEGF/177Lu treatment decreased immunostaining for VEGFR-1, VEGFR-2, and pro-tumorigenic M2-type macrophage marker CD206. Conclusions: Selective targeting of VEGFR with well-tolerated doses of scVEGF/177Lu is effective in metastatic and primary breast cancer models and can be combined with chemotherapy. As high level of VEGFR expression is a common feature in a variety of cancers, targeted delivery of 177Lu for specific receptor-mediated uptake warrants further exploration.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10761
      15. Serial :
        19542
      1. Author :
        Lagerweij, Tonny; Dusoswa, Sophie A.; Negrean, Adrian; Hendrikx, Esther M. L.; de Vries, Helga E.; Kole, Jeroen; Garcia-Vallejo, Juan J.; Mansvelder, Huibert D.; Vandertop, W. Peter; Noske, David P.; Tannous, Bakhos A.; Musters, René J. P.; van Kooyk, Yvette; Wesseling, Pieter; Zhao, Xi Wen; Wurdinger, Thomas
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2017
      5. Publication :
        Angiogenesis
      6. Products :
      7. Volume :
        N/A
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Vasculature; Imaging; 3D; CLARITY; iDISCO; Multicellular network; IVIS; IVIS BLI in vivo
      12. Abstract :
        Three-dimensional visualization of the brain vasculature and its interactions with surrounding cells may shed light on diseases where aberrant microvascular organization is involved, including glioblastoma (GBM). Intravital confocal imaging allows 3D visualization of microvascular structures and migration of cells in the brain of mice, however, with limited imaging depth. To enable comprehensive analysis of GBM and the brain microenvironment, in-depth 3D imaging methods are needed. Here, we employed methods for optical tissue clearing prior to 3D microscopy to visualize the brain microvasculature and routes of invasion of GBM cells.
      13. URL :
        https://link.springer.com/content/pdf/10.1007%2Fs10456-017-9565-6.pdf
      14. Call Number :
        PKI @ catherine.lautenschlager @ 14083
      15. Serial :
        13972
      1. Author :
        Huiling, Liu; Fakhri, Mahdi; Eddie, Perkins
      2. Title :
        Growth factor purification and delivery systems (PADS) for therapeutic angiogenesis
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Vasc Cell.
      6. Products :
      7. Volume :
        7
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Vascular endothelial growth factor; Elastin-like polypeptide; Drug delivery; Therapeutic angiogenesis; Purification and delivery system; IVIS; IVIS FLI ex vivo
      12. Abstract :
        Background Therapeutic angiogenesis with vascular endothelial growth factor (VEGF), delivered either via recombinant protein infusion or via gene therapy, has shown promise in preclinical models of various diseases including myocardial infarction, renovascular disease, preeclampsia, and neurodegenerative disorders. However, dosing, duration of expression, and tissue specificity are challenges to VEGF gene therapy, and recombinant VEGF delivery suffers from extremely rapid plasma clearance, necessitating continuous infusion and/or direct injection at the site of interest. Methods Here we describe a novel growth factor purification and delivery system (PADS) generated by fusion of VEGF 121to a protein polymer based on Elastin-like Polypeptide (ELP). ELP is a thermally responsive biopolymer derived from a five amino acid repeat sequence found in human tropoelastin. VEGFPADS were constructed by fusion of the ELP coding sequence in-frame with the VEGF 121coding sequence connected by a flexible di-glycine linker. In vitroactivity of VEGFPADS was determined using cell proliferation, tube formation, and migration assays with vascular endothelial cells. Pharmacokinetics and biodistribution of VEGFPADS in vivowere compared to free VEGF in mice using quantitative fluorescence techniques. Results ELP fusion allowed for recombinant expression and simple, non-chromatographic purification of the ELP-VEGF 121chimera in yields as high as 90 mg/L of culture and at very high purity. ELP fusion had no effect on the VEGF activity, as the VEGFPADS were equally potent as free VEGF 121in stimulating HUVEC proliferation, tube formation, and migration. Additionally, the VEGFPADS had a molecular weight five-fold larger than free VEGF 121, which lead to slower plasma clearance and an altered biodistribution after systemic delivery in vivo. Conclusion PADS represent a new method of both purification and in vivostabilization of recombinant growth factors. The use of this system could permit recombinant growth factors to become viable options for therapeutic angiogenesis in a number of disease models.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 13683
      15. Serial :
        20764
      1. Author :
        George, Eric M; Liu, Huiling; Robinson, Grant G; Mahdi, Fakhri; Perkins, Eddie; Bidwell III, Gene L
      2. Title :
        Growth factor purification and delivery systems (PADS) for therapeutic angiogenesis
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Vascular cell
      6. Products :
      7. Volume :
        7
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Vascular endothelial growth factor,; Elastin-like polypeptide,; Drug delivery,; Therapeutic angiogenesis,; Purification and delivery system; IVIS; IVIS FLI ex vivo
      12. Abstract :
        Background: Therapeutic angiogenesis with vascular endothelial growth factor (VEGF), delivered either via recombinant protein infusion or via gene therapy, has shown promise in preclinical models of various diseases including myocardial infarction, renovascular disease, preeclampsia, and neurodegenerative disorders. However, dosing, duration of expression, and tissue specificity are challenges to VEGF gene therapy, and recombinant VEGF delivery suffers from extremely rapid plasma clearance, necessitating continuous infusion and/or direct injection at the site of interest. Methods: Here we describe a novel growth factor purification and delivery system (PADS) generated by fusion of VEGF121 to a protein polymer based on Elastin-like Polypeptide (ELP). ELP is a thermally responsive biopolymer derived from a five amino acid repeat sequence found in human tropoelastin. VEGFPADS were constructed by fusion of the ELP coding sequence in-frame with the VEGF121 coding sequence connected by a flexible di-glycine linker. In vitro activity of VEGFPADS was determined using cell proliferation, tube formation, and migration assays with vascular endothelial cells. Pharmacokinetics and biodistribution of VEGFPADS in vivo were compared to free VEGF in mice using quantitative fluorescence techniques. Results: ELP fusion allowed for recombinant expression and simple, non-chromatographic purification of the ELP-VEGF121 chimera in yields as high as 90 mg/L of culture and at very high purity. ELP fusion had no effect on the VEGF activity, as the VEGFPADS were equally potent as free VEGF121 in stimulating HUVEC proliferation, tube formation, and migration. Additionally, the VEGFPADS had a molecular weight five-fold larger than free VEGF121, which lead to slower plasma clearance and an altered biodistribution after systemic delivery in vivo. Conclusion: PADS represent a new method of both purification and in vivo stabilization of recombinant growth factors. The use of this system could permit recombinant growth factors to become viable options for therapeutic angiogenesis in a number of disease models.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 8931
      15. Serial :
        20839
      1. Author :
        Zhou, Heling; Hallac, Rami R; Lopez, Ramona; Denney, Rebecca; MacDonough, Matthew T; Li, Li; Liu, Li; Graves, Edward E; Trawick, Mary Lynn; Pinney, Kevin G
      2. Title :
        Evaluation of tumor ischemia in response to an indole-based vascular disrupting agent using BLI and 19F MRI
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Am J Nucl Med Mol Imaging
      6. Products :
      7. Volume :
        5
      8. Issue :
        2
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Vascular disrupting agents (VDAs),; bioluminescence imaging (BLI),; oximetry,; hypoxia,; 19F MRI,; hexafluorobenzene,; OXi8007,; CA4P; IVIS; IVIS BLI in vivo
      12. Abstract :
        Vascular disrupting agents (VDAs) have been proposed as an effective broad spectrum approach to cancer therapy, by inducing ischemia leading to hypoxia and cell death. A novel VDA (OXi8007) was recently reported to show rapid acute selective shutdown of tumor vasculature based on color-Doppler ultrasound. We have now expanded investigations to noninvasively assess perfusion and hypoxiation of orthotopic human MDA-MB-231/luc breast tumor xenografts following the administration of OXi8007 based on dynamic bioluminescence imaging (BLI) and magnetic resonance imaging (MRI). BLI showed significantly lower signal four hours after the administration of OXi8007, which was very similar to the response to combretastatin A-4P (CA4P), but the effect lasted considerably longer, with the BLI signal remaining depressed at 72 hrs. Meanwhile, control tumors exhibited minimal change. Oximetry used 19F MRI of the reporter molecule hexafluorobenzene and FREDOM (Fluorocarbon Relaxometry using Echo Planar Imaging for Dynamic Oxygen Mapping) to assess pO2 distributions during air and oxygen breathing. pO2 decreased significantly upon the administration of OXi8007 during oxygen breathing (from 122 ± 64 to 34 ± 20 Torr), with further decrease upon switching the gas to air (pO2 = 17 ± 9 Torr). pO2 maps indicated intra-tumor heterogeneity in response to OXi8007, though ultimately all tumor regions became hypoxic. Both BLI and FREDOM showed the efficacy of OXi8007. The pO2 changes measured by FREDOM may be crucial for future study of combined therapy.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 9130
      15. Serial :
        20178
      1. Author :
        Strecker, Tracy E; Odutola, Samuel O; Lopez, Ramona; Cooper, Morgan S; Tidmore, Justin K; Charlton-Sevcik, Amanda K; Li, Li; MacDonough, Matthew T; Hadimani, Mallinath B; Ghatak, Anjan
      2. Title :
        The Vascular Disrupting Activity of OXi8006 in Endothelial Cells and Its Phosphate Prodrug OXi8007 in Breast Tumor Xenografts
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Cancer Letters
      6. Products :
      7. Volume :
        N/A
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Vascular disrupting agent (VDA), M; icrotubules,; Focal adhesion kinase (FAK),; Bioluminescence imaging (BLI),; Breast cancer; IVIS; IVIS BLI in vivo
      12. Abstract :
        This study describes the vascular disrupting ability and the mechanism of action of the indole-based tubulin-binding compound, OXi8006, and its water-soluble phosphate prodrug OXi8007. Treatment of rapidly proliferating human umbilical vein endothelial cells (HUVECs), used as a model for the tumor vasculature, with OXi8006 or OXi8007, caused potent microtubule disruption followed by extensive reorganization of the cytoskeletal network. The mechanism of action involved an increase in focal adhesion formation associated with an increase in phosphorylation of both non-muscle myosin light chain and focal adhesion kinase. These effects were dramatically diminished by an inhibitor of RhoA kinase, a downstream effector of RhoA. Cell cycle blockade at G2/M and cytotoxicity toward rapidly proliferating HUVECs were also observed. Capillary-like networks of HUVECs were disrupted by the action of both OXi8006 and OXi8007. The prodrug OXi8007 exhibited potent and rapid dose-dependent antivascular activity assessed by dynamic bioluminescence imaging (BLI) in an MDA-MB-231-luc breast cancer xenograft mouse model. By 6 hours post treatment, over 93% of the BLI signal was abolished with only a slight recovery at 24 hours. These findings were confirmed by histology. The results from this study demonstrate that OXi8007 is a potent vascular disrupting agent acting through an anti-microtubule mechanism involving RhoA.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10049
      15. Serial :
        20379
      1. Author :
        Jiang, Hai-Fei; Wang, Wei; Jiang, Xuan; Zeng, Wen-Bo; Shen, Zhang-Zhou; Song, Yi-Ge; Yang, Hong; Liu, Xi-Juan; Dong, Xiao; Zhou, Jing; Sun, Jin-Yan; Yu, Fei-Long; Guo, Lin; Cheng, Tong; Rayner, Simon; Zhao, Fei; Zhu, Hua; Luo, Min-Hua
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2017
      5. Publication :
        Journal of Virology
      6. Products :
      7. Volume :
        91
      8. Issue :
        12
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        varicella-zoster virus; ORF7; differentiated neural progenitor cells; differentiated SH-SY5Y cells; cytoplasmic envelopment; Maestro
      12. Abstract :
        Although a varicella-zoster virus (VZV) vaccine has been used for many years, the neuropathy caused by VZV infection is still a major health concern. ORF7 of VZV has been recognized as a neurotropic gene in vivo, but its neurovirulent role remains unclear. In the present study, we investigated the effect of ORF7 deletion on VZV replication cycle at virus entry, genome replication, gene expression, capsid assembly and cytoplasmic envelopment, and transcellular transmission in differentiated neural progenitor cells (dNPCs) and neuroblastoma SH-SY5Y (dSY5Y) cells. Our results demonstrate that the ORF7 protein is a component of the tegument layer of VZV virions. Deleting ORF7 did not affect viral entry, viral genome replication or expression of typical viral genes, but clearly impacted cytoplasmic envelopment of VZV capsids resulting in a dramatic increase of envelop-defective particles and a decrease in intact virions. The defect was more severe in differentiated neuronal cells of dNPCs and dSY5Y. ORF7 deletion also impaired transmission of ORF7-deficient virus among the neuronal cells. These results indicate that ORF7 is required for cytoplasmic envelopment of VZV capsids, virus transmission among neuronal cells and probably the neuropathy induced by VZV infection.IMPORTANCE The neurological damage caused by varicella-zoster virus (VZV) reactivation is commonly manifested as clinical problems. Thus, identifying viral neurovirulent genes and characterizing their functions are important for relieving VZV related neurological complications. ORF7 has been previously identified as a potential neurotropic gene, but its involvement in VZV replication is unclear. In this study, we found that ORF7 is required for VZV cytoplasmic envelopment in differentiated neuronal cells, and the envelopment deficiency caused by ORF7 deletion results in poor dissemination of VZV among neuronal cells. These findings imply that ORF7 plays a role in neuropathy, highlighting a potential strategy to develop a neurovirulence-attenuated vaccine against chickenpox and herpes zoster, and providing a new target for intervention of neuropathy induced by VZV.
      13. URL :
        http://jvi.asm.org/content/early/2017/03/23/JVI.00127-17.abstract http://jvi.asm.org/content/91/12/e00127-17.full.pdf
      14. Call Number :
        PKI @ catherine.lautenschlager @ 13607
      15. Serial :
        12849
      1. Author :
        Zhang, Zhen; Huang, Ying; Zhu, Hua
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        In Vitro Mutagenesis Protocols
      6. Products :
      7. Volume :
        634
      8. Issue :
        N/A
      9. Page Numbers :
        75
      10. Research Area :
        N/A
      11. Keywords :
        Varicella-zoster virus, Bacterial artificial chromosome, Deletion mutagenesis, Bioluminescence IVIS, Xenogen
      12. Abstract :
        Varicella-zoster virus (VZV) causes both varicella (chicken pox) and herpes zoster (shingles). As a member of the human herpesvirus family, VZV contains a large 125-kb DNA genome, encoding 70 unique open reading frames (ORFs). The genetic study of VZV has been hindered by the large size of viral genome, and thus the functions of the majority of these ORFs remain unclear. Recently, an efficient protocol has been developed based on a luciferase-containing VZV bacteria artificial chromosome (BAC) system to rapidly isolate and study VZV ORF deletion mutants.
      13. URL :
        http://www.springerprotocols.com/Abstract/doi/10.1007/978-1-60761-652-8_5
      14. Call Number :
        145084
      15. Serial :
        5383
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