Vascular Biology Research Research Articles

Utilizing Zebrafish to Identify Anti-(Lymph)Angiogenic Compounds for Cancer Treatment: Promise and Future Challenges.

Cancer metastasis which predominantly occurs through blood and lymphatic vessels, is the leading cause of death in cancer patients. Consequently, several anti-angiogenic agents have been approved as therapeutic agents for human cancers such as metastatic renal cell carcinoma. Also, anti-lymphangiogenic drugs such as monoclonal antibodies VGX-100 and IMC-3C5 have undergone phase I clinical trials for advanced and metastatic solid tumors. Although anti-tumor-associated angiogenesis has proven to be a promising therapeutic strategy for human cancers, this approach is fraught with toxicities and development of drug resistance. This emphasizes the need for alternative anti-(lymph)angiogenic drugs. The use of zebrafish has become accepted as an established model for high-throughput screening, vascular biology, and cancer research. Importantly, various zebrafish transgenic lines have now been generated that can readily discriminate different vascular compartments. This now enables detailed invivo studies that are relevant to both human physiological and tumor (lymph)angiogenesis to be conducted in zebrafish. This review highlights recent advancements in the zebrafish anti-vascular screening platform and showcases promising new anti-(lymph)angiogenic compounds that have been derived from this model. In addition, this review discusses the promises and challenges of the zebrafish model in the context of anti-(lymph)angiogenic compound discovery for cancer treatment.

The Future of Vascular Biology and Medicine.

During the past several decades, landmark discoveries in the field of vascular biology have evolved our understanding of the biology of blood vessels and the pathobiology of local and systemic vascular disease states and have led to novel disease-modifying therapies for patients. In 1980, Furchgott and Zawadzki1 radically changed our focus in blood vessel research with the discovery of endothelium-derived nitric oxide and its effects on vascular tone. This seminal discovery redefined blood vessels as dynamic organs with autocrine, paracrine, and endocrine functions that are capable of regulating their own environment. The later recognition that many risk factors associated with vascular disease perturb nitric oxide-mediated homeostatic mechanisms lent further credence to the concept of blood vessels as complex organs rather than inert tubes. In the decades to follow, research in vascular biology predominantly targeted 3 facets of vascular function: vasomotor tone, inflammation, and the balance between thrombosis and thrombolysis. This occurred because atherosclerotic cardiovascular disease reached epidemic levels and atherothrombosis was found to feature disturbances in these functions that preceded visible pathology and clinical manifestations of the disease. Furthermore, modification of responsible causal factors reversed impaired vascular function (eg, lowering levels of low-density lipoproteins in atherosclerosis), and clinical studies began to validate the importance of preclinical vascular biology research in the treatment of hypertension, atherosclerosis, pulmonary vascular disease, erectile dysfunction, Raynaud phenomenon, and neointimal proliferation after mechanical vascular intervention. More recently, advances in molecular biology and –omics technologies have facilitated in vitro and in vivo studies that revealed that blood vessels regulate their own redox milieu, metabolism, mechanical environment, and phenotype, in part, through complex interactions between cellular components of the blood vessel wall and circulating factors. These interactors include stem, progenitor, and differentiated cells; microRNAs, long noncoding RNAs, and DNA; and, hormones, proteins, and lipids. Dysregulation of these …

Molecular Imaging of Angiogenesis and Vascular Remodeling in Cardiovascular Pathology.

Angiogenesis and vascular remodeling are involved in a wide array of cardiovascular diseases, from myocardial ischemia and peripheral arterial disease, to atherosclerosis and aortic aneurysm. Molecular imaging techniques to detect and quantify key molecular and cellular players in angiogenesis and vascular remodeling (e.g., vascular endothelial growth factor and its receptors, αvβ3 integrin, and matrix metalloproteinases) can advance vascular biology research and serve as clinical tools for early diagnosis, risk stratification, and selection of patients who would benefit most from therapeutic interventions. To target these key mediators, a number of molecular imaging techniques have been developed and evaluated in animal models of angiogenesis and vascular remodeling. This review of the state of the art molecular imaging of angiogenesis and vascular (and valvular) remodeling, will focus mostly on nuclear imaging techniques (positron emission tomography and single photon emission tomography) that offer high potential for clinical translation.

Sphingosine-1-Phosphate Signaling in Endothelial Disorders.

Numerous preclinical studies indicate that sustained endothelial activation significantly contributes to tissue edema, perpetuates the inflammatory response, and exacerbates tissue injury ultimately resulting in organ failure. However, no specific therapies aimed at restoring endothelial function are available as yet. Sphingosine-1-phosphate (S1P) is emerging as a potent modulator of endothelial function and endothelial responses to injury. Recent studies indicate that S1PR are attractive targets to treat not only disorders of the arterial endothelium but also microvascular dysfunction caused by ischemic or inflammatory injury. In this article, we will review the current knowledge of the role of S1P and its receptors in endothelial function in health and disease, and we will discuss the therapeutic potential of targeting S1PR not only for disorders of the arterial endothelium but also the microvasculature. The therapeutic targeting of S1PR in the endothelium could help to bridge the gap between biomedical research in vascular biology and clinical practice.

Clinical and biochemical characteristics are varied in subjects with ABCA1 mutations

s / Atherosclerosis 241 (2015) e32ee71 e37 other fragments of exon 26 and 29 as well as in exon 3 and 22 have been reported in FH patients. The main aim of this project was to functional characterize novel alterations in exon 19 and 26 (APOB) found in clinical FH patients. Methods: LDL from 2 index patients and relatives was separated and marked with FITC-LDL to perform functional studies by flow cytometry with lymphocytes and HepG2, and U937 growth assays. Alterations were screened in a 96 normolipidaemic panel by PCR and sanger sequencing. Results: In vitro analysis of these variants showed a decrease in the binding and internalization of LDL and in U937 growth from individuals with exon 26 alteration, showing a similar effect as APO3527. As reported before also in this family the penetrance is also reduced. Alteration in exon 19 had a neutral effect. Both alterations were not present in the normolipidaemic panel. Conclusions: The spectrum of functional alterations in APOB outside the fragments routinely screened is growing. Screening of all 29 exons of APOB should be performed in routine diagnosis, now possible by NGS, since it is expected that a further 10% of clinical FH patients can have FH due to a novel APOB mutation. If so mutations in APOB can be a common cause of FH than expected. EAS-0599. CLINICAL AND BIOCHEMICAL CHARACTERISTICS ARE VARIED IN SUBJECTS WITH ABCA1 MUTATIONS M. Cuchel , J.T. McParland , J. Wang , R.A. Hegele , D.J. Rader . Department of Medicine Division of Translational Medicine and Human Genetics, University of Pennsylvania, Philadelphia, USA; Vascular Biology Research Group, Robarts Research Institute, London, Canada Aim: The ATP-binding cassette transporter 1 (ABCA1) is a membrane protein responsible for generating HDL through its ability to efflux phospholipid and cholesterol and for maintaining cellular cholesterol homeostasis. Recent data suggest that ABCA1 may affect other metabolic pathways beyond HDL metabolism. The clinical and biochemical characterization of subjects carrying mutations in the gene encoding for ABCA1 may help gain some insights into the frequency and pathophysiology of this condition. Methods: We performed a detailed lipid profile and ABCA1 sequencing in subjects with primary hypoalphalipoproteinemia. Where possible, family members of subjects carrying ABCA1 mutations were also studied. Results: ABCA1 mutations were found in 25 of 49 probands sequenced. Additional carriers were found among family members. Homozygous or compound heterozygous subjects (n1⁄413, age 30±12 years) had HDL-cholesterol1⁄46.5±8.0 mg/dl (median 4, range 1-31 mg/dl) and mean apoA-I1⁄4 21±31 mg/dl. Of these 13 subjects, 3 (23%) had enlarged tonsils, 2 (15%) reported neuropathy, 1 (8%) had clinically evident CAD, and none had diabetes. Heterozygous subjects (n1⁄431, age 39±18 years) had HDL-cholesterol1⁄422±9 mg/dl and apoA-I1⁄4 80±37 mg/dl. Of these, 4 (13%) had diabetes, and 2 (6%) had clinically evident CAD. Conclusions: ABCA1mutations may be amore common cause of lowHDLcholesterol than is appreciated. Additionally, these data highlight the broad spectrum of HDL-cholesterol and apoA-I levels as well as clinical characteristics that are found in carriers of ABCA1 mutations. EAS-0660. THE CARDIOVASCULAR GENE ANNOTATION INITIATIVE: IMPACT ON DATA ANALYSIS R.C. Lovering , M. Rodriguez-Lopez , N.H. Campbell , R.P. Huntley , T. Sawford , C. O’Donovan , S. Orchard , H. Hermjakob , M. Martin , M. Mayr , S.E. Humphries , P.J. Talmud . 1 Institute of Cardiovascular Science, UCL, London, United Kingdom; UniProt development, European Molecular Biology Laboratory European Bioinformatics Institute, Hinxton, United Kingdom; UniProt content, European Molecular Biology Laboratory European Bioinformatics Institute, Hinxton, United Kingdom; 4 Proteomics, European Molecular Biology Laboratory European Bioinformatics Institute, Hinxton, United Kingdom; Cardiovascular, King's College London, London,

Vascular Calcification - Pathological Mechanism and Clinical Application - . Vascular calcification in aged mice

Vascular calcification is a risk factor for cerebral and cardiovascular events and has a high prevalence among elderly. To finding ways of prevent or cure vascular calcification may leads to not only a healthy longevity but also medical expenditure reduction. However, the molecular mechanism underlying this pathogenic process is still obscure. To clarify the mechanism of vascular calcification, the development of animal models that exhibit extensive and robust vascular calcification is an important issue for research in vascular biology.

The Society for Vascular Medicine: the first quarter century.

The Society for Vascular Medicine was founded in 1989. During the subsequent 25 years, the Society has grown to approximately 500 members and has achieved international recognition while making important contributions to vascular disease education, clinical vascular medicine and biology research, and patient care. In celebration of the Society's 25th anniversary, its past and current presidents reflect on the Society's history, challenges, and achievements, and emphasize the vital role of the SVM in the discipline of vascular medicine.

Vascular niches: endothelial cells as tissue- and site-specific multifunctional team players in health and disease.

The cardiovascular system is a circulatory system that transports the blood and reaches out to almost every site of the body. Endothelial cells (EC) as the innermost cell layer of blood vessels constitute a huge interface between the blood and nearly all other tissues and cells. Although blood vessels and their EC were considered as rather inert and passive conduits early in the history of vascular discoveries, progress in vascular research now suggests that EC are actively involved in a plethora of physiological or pathophysiological processes. Endothelial heterogeneity and the concept of vascular niches are in the spotlight of current research in vascular biology. Endothelial heterogeneity comprises morphologic, molecular and functional features. It allows EC to adapt to the organ- and site-specific requirements exhibited by different segments of the vascular tree. In organ-specific vascular niches, EC mutually interact with neighboring cells. EC-derived cytokines, called angiokines, acting in a paracrine (angiocrine) fashion have been shown to be decisively involved in various biological processes ranging from stem cell maintenance to organ regeneration as well as cancer progression and metastasis. In summary, EC should always be considered as teammates in the maintenance and regulation of complex multicellular network interactions in health and disease.

Creating perfused functional vascular channels using 3D bio-printing technology

We developed a methodology using 3D bio-printing technology to create a functional in vitro vascular channel with perfused open lumen using only cells and biological matrices. The fabricated vasculature has a tight, confluent endothelium lining, presenting barrier function for both plasma protein and high-molecular weight dextran molecule. The fluidic vascular channel is capable of supporting the viability of tissue up to 5 mm in distance at 5 million cells/mL density under the physiological flow condition. In static-cultured vascular channels, active angiogenic sprouting from the vessel surface was observed whereas physiological flow strongly suppressed this process. Gene expression analysis was reported in this study to show the potential of this vessel model in vascular biology research. The methods have great potential in vascularized tissue fabrication using 3D bio-printing technology as the vascular channel is simultaneously created while cells and matrix are printed around the channel in desired 3D patterns. It can also serve as a unique experimental tool for investigating fundamental mechanisms of vascular remodeling with extracellular matrix and maturation process under 3D flow condition.

Utilizing PCL Microcarriers for High-Purity Isolation of Primary Endothelial Cells for Tissue Engineering

Endothelial cells (ECs) are widely used in research, both for fundamental vascular biology research and for exploring strategies to create engineered vascularized tissues. Primary isolation often results in contamination from fibroblasts and vascular smooth muscle cells that can potentially affect function, particularly during the initial expansion period needed to establish the cell culture. In the current study, we explored the use of microcarriers to selectively isolate ECs from the lumen of intact vessels to enhance the purity during the isolation procedure. First, rat aortic explant culture was performed and after 2 weeks of culture, flow cytometry revealed that only 60% of the expanded cell population was positive for the endothelial marker CD31. Then, we employed a strategy to selectively isolate ECs and improve their purity by introducing microcarriers to the lumen of intact aorta. After 10 days, microcarriers were carefully removed and placed in cell culture dishes and at 15 days, a large near confluent layer of primary ECs populated the dish. Flow cytometry revealed that >90% of the expanded cells expressed CD31. Moreover, the cells were capable of forming tubule-like structures when plated onto Matrigel, confirming their function also. The highly modular and transportable nature of microcarriers has significant potential for isolating ECs at high purity, with minimal contamination.

Novel Therapeutic Paradigms to Treat Hemodialysis Access Dysfunction

Vascular access dysfunction continues to be a major factor contributor to the morbidity and mortality of hemodialysis patient. Percutaneous intervention has been the central therapeutic modality used to treat dialysis vascular access dysfunction with limited success. Vascular biology research has shed considerable light on the pathophysiologic processes that lead to the development of neointimal hyperplasia, the culprit lesion for vascular access dysfunction, suggesting possible newer novel therapeutic interventions. In this review we discuss; (1) recent advances in the utilization of image based predictive medicine in improving access type selection prior to access creation; (2) current and promising treatment modalities including brachytherapy, gene therapy and pharmacotherapy for prevention of neointimal hyperplasia; (3) alternate imaging modalities during percutaneous endovascular interventions. Though novel therapeutic interventions are evolving, robust clinical studies to identify optimal therapeutic method are needed. A combination of evidenced based interventions from pre access creation up until final abandonment of vascular access is worthy of exploration.

Go with the flow: The passion for vascular biology research

Go with the flow: The passion for vascular biology research

Visualization of Endothelial Actin Cytoskeleton in the Mouse Retina

Angiogenesis requires coordinated changes in cell shape of endothelial cells (ECs), orchestrated by the actin cytoskeleton. The mechanisms that regulate this rearrangement in vivo are poorly understood - largely because of the difficulty to visualize filamentous actin (F-actin) structures with sufficient resolution. Here, we use transgenic mice expressing Lifeact-EGFP to visualize F-actin in ECs. We show that in the retina, Lifeact-EGFP expression is largely restricted to ECs allowing detailed visualization of F-actin in ECs in situ. Lifeact-EGFP labels actin associated with cell-cell junctions, apical and basal membranes and highlights actin-based structures such as filopodia and stress fiber-like cytoplasmic bundles. We also show that in the skin and the skeletal muscle, Lifeact-EGFP is highly expressed in vascular mural cells (vMCs), enabling vMC imaging. In summary, our results indicate that the Lifeact-EGFP transgenic mouse in combination with the postnatal retinal angiogenic model constitutes an excellent system for vascular cell biology research. Our approach is ideally suited to address structural and mechanistic details of angiogenic processes, such as endothelial tip cell migration and fusion, EC polarization or lumen formation.

Endothelium: from cellophane to orchestral maestro

Endothelial cells from human umbilical veins were first cultured nearly four decades ago, initiating explosive growth in research in vascular biology and leading to major insights into angiogenesis, vasculogenesis, and tumor biology. Recent studies now promise to open new horizons in regenerative medicine as well as organ engineering.

A Suggested Roadmap for Cardiovascular Ultrasound Research for the Future

Sanjiv Kaul, MD, FASE,* James G. Miller, PhD,* Paul A. Grayburn, MD, Shinichi Hashimoto,Mark Hibberd, MD, PhD, Mark R. Holland, PhD, FASE, Helene C. Houle, BA, RDMS, RDCS, RVT, FASE,Allan L. Klein, MD, FASE, Peg Knoll, RDCS, FASE, Roberto M. Lang, MD, FASE,Jonathan R. Lindner, MD, FASE, Marti L. McCulloch, RDCS, FASE, Stephen Metz, PhD,Victor Mor-Avi, PhD, FASE, Alan S. Pearlman, MD, FASE, Patricia A. Pellikka, MD, FASE, Nancy DeMarsPlambeck,BS,RDMS,RDCS,RVT,DavidPrater,MS, ThomasR.Porter,MD,FASE,DavidJ.Sahn,MD,FASE,James D. Thomas, MD, FASE, Kai E. Thomenius, PhD, and Neil J. Weissman, MD, FASEINTRODUCTIONThe leadership at the American Society of Echocardiography (ASE)decided on a proactive role in defining selected areas of researchnecessary in this decade that will meet our future clinical needs.Consequently, ASE sponsored a Technology and ResearchSummit in the fall of 2010 in conjunction with the AmericanHeart Association Scientific Sessions in Chicago. In addition to theASE executive committee, in attendance were the editor, deputyeditor, and one of the associate editors of the Journal of theAmerican Society of Echocardiography. Also invited were physician-scientists active in the field of cardiovascular ultrasound, respectedultrasound physicists, and senior engineers from the various ultra-sound companies.The agenda for the full-day meeting covered a selected range ofsubjects including the assessment of global and regional left ventricu-lar function, regional myocardialperfusion, molecular imaging, thera-peutic ultrasound, and peripheral vascular imaging. Also addressedwere research necessary to determine the broad clinical utility ofhand held ultrasound devices and the impact of future technologicaldevelopments on the field of cardiovascular imaging.Because of time constraints, other important and worthy areas ofresearch were not discussed. There was an hour devoted to the dis-cussion of each subject that was initiated by the chairs and panelistsassigned to each of the topics. The discussion was robust, and at theend, the chairs and panelists for each topic were requested to sub-mit in writing a short synopsis of the discussion. These have beencompiled into a document that we believe will serve as a roadmapfor cardiovascular ultrasound research for this decade. At the endof each section a short list of references for selected reading isprovided.Although we have defined the areas that are ripe for future re-search, we also strongly believe that we havetotrain the future scien-tists who will implement this research agenda. ASE has historicallyawarded one or two fellowship training grants a year and also anaward for researchtraining of a sonographer. At some institutions fel-lowshavealsoreceivedtraininggrantsfromthelocalAmericanHeartAssociation, and very occasionally a training grant (F32) from theNational Institutes of Health. However, this is not enough. We needmore institutional training grants from the National Institutes ofHealth in order to train an adequate number of MD and PhD scien-tists in cardiovascular imaging. To our knowledge there are currentlyonlyahandfulofsuchtraininggrantsinthecountry,whichiswoefullyinadequate. We believe that we need at least 20–25 such traininggrants devoted to the general field of cardiovascular imaging so thatwithin a decade there will be enough physicians trained in scientificmethods and clinical research to address the subjects that havebeen discussed in this report.The field of cardiovascular ultrasound is very broad, ranging fromclinical validation of new technology to studies requiring knowledgeof physics, mathematics, organic chemistry, physiology, pharmacol-ogy, molecular and vascular biology, genetics, clinical trials, and out-come research. Cross-training of individuals in one or more of thesefields is essential for cardiovascular ultrasound to thrive and succeed.Ourhopeisthatthisreportwillencourageyoungpeopletorealizethescope of cardiac ultrasound research and make a career in this dy-namic field.Selected Reading

Frontiers of vascular biology and disease research

2010 represents a special year for Acta Pharmacologica Sinica, the 30th anniversary of its debut as an international journal. On this memorable occasion, the journal and its guest editors are proud to present to our readers a special issue in vascular biology and disease research. The research work presented herewith spans from molecular cardiovascular biology to integrated physiology and pharmacology.

Strategic Plan for Lung Vascular Research

The Division of Lung Diseases of the National Heart, Lung, and Blood Institute, with the Office of Rare Diseases Research, held a workshop to identify priority areas and strategic goals to enhance and accelerate research that will result in improved understanding of the lung vasculature, translational research needs, and ultimately the care of patients with pulmonary vascular diseases. Multidisciplinary experts with diverse experience in laboratory, translational, and clinical studies identified seven priority areas and discussed limitations in our current knowledge, technologies, and approaches. The focus for future research efforts include the following: (1) better characterizing vascular genotype-phenotype relationships and incorporating systems biology approaches when appropriate; (2) advancing our understanding of pulmonary vascular metabolic regulatory signaling in health and disease; (3) expanding our knowledge of the biologic relationships between the lung circulation and circulating elements, systemic vascular function, and right heart function and disease; (4) improving translational research for identifying disease-modifying therapies for the pulmonary hypertensive diseases; (5) establishing an appropriate and effective platform for advancing translational findings into clinical studies testing; and (6) developing the specific technologies and tools that will be enabling for these goals, such as question-guided imaging techniques and lung vascular investigator training programs. Recommendations from this workshop will be used within the Lung Vascular Biology and Disease Extramural Research Program for planning and strategic implementation purposes.

Three-dimensional cell-dense constructs containing endothelial cell-networks are an effective tool for in vivo and in vitro vascular biology research

Angiogenesis is a complicated natural process, and understanding the mechanism by which it occurs is important for medical, pharmaceutical, and cell biological sciences. Many techniques for investigating angiogenesis have been reported. In this study, we introduced a novel application of a cell culture technique that can be used in in vitro and in vivo vascular biology research. Cultivated endothelial cells (ECs) were harvested from temperature responsive culture dishes by reducing the temperature, without the need for a proteinase treatment. For this technique, the direct contact of ECs with fibroblasts was important for the formation of a capillary-like network in vitro. Moreover, layered cell sheets containing EC-networks produced lumen and vascular structures in the three-dimensional constructs, as well as in the construct transplanted into a living body. Thus, our culture technique was able to create cell sheets and three-dimensional constructs containing EC-networks, because they preserved normal and intrinsic cell-cell direct contact and various cell adhesive factors. Moreover, the thickness of these three-dimensional (3-D) constructs could be controlled by the number of layered cell sheets. These observations indicated that our novel technology contributed to the progress of vascular biology and lead to a new tool that can be used in in vivo and in vitro vascular biology research.

IT & informatics in translational research: a case study

IT & informatics in translational research: a case study

Variant type Glanzmann thrombasthenia caused by homozygous p.724R > X mutation in β 3 integrin

a Clinical Research Center, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary b Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary c Pal Heim Children's Hospital, Budapest, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary d Department of Obstetrics and Gynecology, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary e Clinical Biochemistry and Molecular Pathology Department, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary