Developmental Vascular Biology Program

Current events/important calendars

Vascular Biology Interest Group

Developmental Vascular Biology Program Presentation Schedule

Leadership

Contact information:
8701 Watertown Plank Road
Milwaukee, WI 53226

Ramchandran lab:

	Chang Zoon Chun, PhD, research scientist I - My research focuses on how endothelial precursor cells, angioblasts differentiate into primary axial vessels during embryonic development in vertebrates. Research interests include studying fundamental vasculogenesis process and its contribution to vasculogenic diseases. cchun@mcw.edu (414) 955-2388
	Chris Koceja, technician - My research entails combining two different transgenic approaches to over-express oncogenes in sympathetic neurons of the zebrafish nervous system, while utilizing the GAL4/UAS system. ckoceja@mcw.edu (414) 456.5896
	Noah Leigh, MS, research associate I- My research interests are focused on drug discovery related to the process of angiogenesis. Currently I am working on the characterization of a novel gene DU644441 that I worked on for my Masters thesis.  My goal is to determine the cell type the gene is expressed in during embryonic development in zebrafish and to confirm this gene is a matrix metalloproteinase (MMP) by conducting MMP assays using expressed protein.   nleigh@mcw.edu (414) 955-2392
	Keguo Li, PhD, post-doctoral fellow - My research interest lies in understanding the significance of intergenic transcripts. Specifically, I am interested in the roles of the transcripts in the flanking regions of Dll4 locus. We use quantitative visualizing technology with single molecular sensitivity to track such transcripts during vascular development. Gain- and loss-of-function studies are utilized to probe the impact of the transcripts on angiogenesis. Another intriguing field to me is the positional cloning of subtelomeric genes. kli@mcw.edu (414) 456-7356
	Marcus Schupp, PhD, post-doctoral fellow - My work aims to unravel molecular mechanisms controling the patterning and cellular behavior of vascular progenitor cell populations during early steps of vertebrate vasculogenesis. In particular, by using zebrafish transgenesis and imaging technology, I will investigate the gene-regulatory networks underlying the specification and differentiation of vasculogenic precursors and investigate signaling pathways affecting their proliferative and migratory properties. mschupp@mcw.edu (414) 955-2557
	Ling Wang, PhD, post-doctoral fellow - xxxxx. lingwang@mcw.edu (414) 955-2390
	Srividya Suryanarayana, PhD, post-doctoral fellow - My project focuses on understanding the role of specific components in the MAP kinase pathway. MAPKs play an important role in hemangiomas and vascular anamolies and are therefore promising drug targets. More specifically, using biochemical approaches, my current work involves studying the interaction of DUSP5 (a dual-specificity protein MAPK phosphatase) with ERKs (Extracellular signal-regulated kinases) 1 and 2. My long-term interest is to develop a high throughput assay for small molecule screening on the target Dusp-5. vsuryanaraya@mcw.edu (414) 955-2397
	Padmanabhan Vakeel, PhD, post-doctoral fellow - Kinases and phosphatases play a major role in all cellular functions of the cell and have wider implications in the disease state. My investigations are focused on understanding their intricate connection during vasculogenesis and angiogenesis using model systems such as zebrafish, mouse, and mammalian cell culture. The GOF/LOF (Gain/Loss of function) studies on these molecules in model systems would reveal the spatial and temporal nature of their function within the complex cellular signaling network.pvakeel@mcw.edu (414) 955-2963
	Anil Challa, PhD, post-doctoral fellow - I am interested in understanding the molecular mechanisms that control cell motility and changes in cell morphology in various developmental processes during vertebrate embryogenesis. I have a special interest in the biological functions of the Roundabout (Robo) family of receptor proteins, which have been implicated in phenomena such as axon guidance and vasculogenesis. achalla@mcw.edu (414) 955-2274

Wilkinson Lab:

Indu Remadevi, MS, research associate I - My primary interests are in studying the function of early markers associated with vascular development in zebrafish. iremadev@mcw.edu (414) 955-2219 Sreenivas Kilari, PhD, post-doctoral fellow - At present, I have two research interests based on my current projects. In the first project, I am focusing on the VEGFR-2/KDR signaling as modified by ECSCR (Endothelial Cell Specific Chemotaxis Receptor) during vascular development. In a second project, I am examining ephrin-B2 signaling partners in endothelial cells. skilari@mcw.edu (414) 955-2273.

The Developmental Vascular Biology program investigates the basic mechanisms of blood vessel formation in vertebrates and the contribution of the vasculature to disease states. We study how a specified endothelial precursor cell (angioblast) eventually differentiates into artery or vein. The basic mechanisms of this process often are dysregulated in disease. Therefore, understanding the basic mechanisms of vessel formation will generate new treatments for conditions affected by deregulated vessel growth. In particular, vascular conditions associated with children such as hemangiomas and solid tumors will benefit because ongoing studies in this program utilize developing zebrafish and mouse embryos. Zebrafish studies specifically are primarily embryonic in nature and thus directly contribute to child development. In addition, tools for performing drug screens using zebrafish embryos are being developed, which will identify targets and potential drug leads for treating pediatric vascular conditions.

During development blood vessels adapt to specific needs of the organ they supply. One highly vascularized organ system is the lungs. In the lungs, the vasculature becomes highly specialized to provide efficient gas-exchange by participating with alveoli. Development of lung alveoli requires complex interactions among blood vessels, other specialized cells and the extracellular matrix. Defects in any one of these elements will adversely affect alveolar development. Human lung pathologies including emphysema and infant bronchopulmonary dysplasia are associated with defective alveolar structure and concomitant vascular pathologies. 

Other research projects

Ramchandran lab:

• Angioblast development in vertebrates
  

  Endothelial precursor cells or angioblasts are specified from lateral mesoderm cells in the developing vertebrate embryo. The intermediate steps necessary for the angioblast to form arteries and veins are not clear. The Ramchandran laboratory studies the signals and processes involved in the different steps of angioblast development in a developing embryo using a variety of cell biology, genetic and molecular approaches.

• Role of axon guidance genes in vascular development
  

  Vessels and nerves are branching networks that often lay side by side in a developing embryo. Recently, mechanisms governing branching morphogenesis are shared both at the cell surface and the intracellular levels in endothelial cells and axons respectively. The Ramchandran lab is interested in understanding the molecular mechanisms that guide an endothelial cell to its target using clues from the axon guidance system. We study one member of the axon guidance family, roundabout4 (robo4), and its role in this process

• Translational disease models and Drug Discovery

  Vessel growth is tightly regulated during development. Any imbalance in this regulation often is associated with disease. For example, tumor growth is dependent on neo-vessel growth or angiogenesis. The Ramchandran laboratory is developing disease models in fish using a variety of genetic and molecular approaches that will eventually be used for drug screening. We have recently launched computational small molecule and conventional screening approaches to target mutant protein vs. wild type protein in the vasculature. Using such approaches we have identified candidate small molecules against targets mutated in human vascular anomaly disease. The efficacy of some compounds is in the nanomolar range, which is being further developed as therapeutics. In addition, the Ramchandran lab has taken a leading role in forming a drug discovery consortium in Southeastern Wisconsin with the focus on merging individual expertise across participating institution laboratories in SE Wisconsin towards a common goal of generating affordable therapeutics that will benefit the health of the local communities and beyond.

Wilkinson lab:

• Cell surface receptors affecting endothelial migration Angiogenesis, the formation of new vessels from a preexisting network, depends proper mobilization of blood endothelial cells. We are using a combination of molecular genetic and cell biology approaches to study cell-surface control of endothelial cell motility. In our first project, we are studying the impact of endothelial ephrin-B2 in lung alveolar development. The abnormal lung development of mice expressing mutant ephrin-B2 provides an opportunity to investigate the phenomenon of alveolar initiation from a cell and developmental biology point of view. In a second project, we are evaluating a poorly understood novel cell-surface protein ECSCR, which is selectively expressed in endothelial cells and influences endothelial migration and proliferation

List of Articles Presented

Important Research Technique/Methodology Papers

The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments Stephen A. Bustin,1* Vladimir Benes,2 Jeremy A. Garson,3,4 Jan Hellemans,5 Jim Huggett,6 Mikael Kubista,7,8 Reinhold Mueller,9 Tania Nolan,10 Michael W. Pfaffl,11 Gregory L. Shipley,12 Jo Vandesompele,5 and Carl T. Wittwer13,14

Generation of Transgenic Zebrafish Using Tol2 Thomas S. Becker Lab, Pavla Navratilova

ZF Sectioning Nature Protocols  Jessica Sullivan-Brown, Margaret E. Bisher & Rebecca D. Burdine

Resource links

Resources for gene-specific reagents

Public aggregator sites

1. The NCBI Gene Page provides annotation including PubMed links and (at the bottom) aggregates cDNA, small nucleotide and antiserum links.

2. Bioinformatics Harvester gathers data from a number of gene pages, protein predictions and also links to products related to specific genes. Entry is via text search and is not very good with alternate gene names.

Commercial aggregators

1. Biocompare lists antisera, short nucleotide reagents and small molecules from about 20 vendors. Searchable by text, does not typically recognize alternate names.
2. Exact Antigen (To be renamed Labome in 2010) lists antisera, short nucleotide reagents and cDNA clones from multiple vendors. Text search is very robust and will identify alternative names for gene products.
3. Origene sells expression constructs including C-terminal fusion constructs for many human and mouse gene products. Also sells short nucleotide reagents.

Individual vendors

1. Open Biosystems has a very large set of cDNAs and nucleotide reagents. Searchable by text or BLAST.
2. Invitrogen sells a large number of gene-specific reagents.
3. Genecopoeia sells a wide variety of gene-specific reagents, including Gateway-ready entry constructs.

Mutant allele collections

1. ITGC, the International Gene Trap Consortium, lists a large number of mouse ES cell lines with gene trap integration intended to generate null alleles.
2. The Sanger Institute Zebrafish Mutation Resource generates zebrafish mutant lines by ENU mutagenesis followed by exon sequencing recovery. Users can request a gene to be targeted with delivery time of up to 12 months.

Miscellaneous

1. http://www.fishnet.org.au/  This site has optical position tomography images of zebrafish sections.  Use this site to refer to anatomical location of the structure that you are interested in identifying.
2. http://www.anaspec.com/products/productcategory.asp?id+857  This company sells a few antisera against zebrafish antigens.  They also sell de-yolked lysates, probably by a similar protocol to ours.  The name of the company is Eurogentec Anaspec.

Awards

• 2002-2008
  National Institutes of Health NCI Scholar Award, Ramani Ramchandran, PhD
• 2007-2010
  Advancing a Healthier Wisconsin Cloche: A basic and translational model for cardiovascular disease
• 2008
  Angioblast development in zebrafish development, Young Investigator Award, Mayo Clinic Angiogenesis Symposia, Chang Zoon Chun, PhD
• 2008
  EphrinB2 role in mouse lung vascular development, Young Investigator Award, 3rd Mayo Clinic Angiogenesis Symposia, George Wilkinson, PhD
• 2010-2011
  Identification of the physiologically relevant ligand for the angiogenic roundabout (Robo-4) receptor, American Heart Association Midwest Affiliate, Ganesh Samant, PhD

Invited presentations 2010-present

Intramural--(MCW)

• 2010 "Functional characterization of Snrk-1 gene in developing vasculature – an update"- CVRC Works-in-Progress
• 2010 "Vasculogenesis mechanisms in vertebrates" - CBNA Presentation
• 2010 "How to make sense of anti-sense RNA" - CRI Noon Conference
• 2010 "Early mechanistic steps of vasculogenesis process revealed in zebrafish" - June Multidisciplinary Research Conference
• 2010 "Successful strategies for science and life" - Summer Research Lecture Series
• 2011 "Early mechanistic steps of vasculogenesis process revealed in zebrafish" - Pediatric Surgery Multidisciplinary Research Conference
• 2011 Small Molecule Identification for a Vascular Target: Dusp-5. Pediatric Surgery Multidisciplinary Research Conference, Medical College of Wisconsin, Milwaukee, WI
• 2011 Bridging the gap from targets to drugs (Valley of Death):  A proposal for a SE WI Drug Development Core.  Medical College of Wisconsin, Milwaukee, WI
• 2011 Targeting Dusp-5:  An emerging option for treating vascular anomalies.  CTSI Seminar Presentation, Medical College of Wisconsin, Milwaukee, WI
• 2011 Insights on Aortic Coarctation Defects from Studying Embryonic Vasculogenesis Process in Zebrafish.  Children's Research Institute, Medical College of Wisconsin, Milwaukee, WI

Extramural--

• 2010 "Vascular Development in Vertebrates" - Physiology and Biophysics, University of Louisville School of Medicine, Louisville, KY
• 2010 "Vasculogenesis mechanisms in a developing vertebrate" -  Cardiovascular Seminar Series, University of Virginia Health System, Charlottesville, VA
• 2010 "Nogo-B receptor is essential for angiogenesis in vivo via Akt pathway" -  American Heart Association, San Francisco, CA
• 2010 "Developmental vasculogenesis mechanisms during embryogenesis and disease in vertebrates" - Beihang University of Aeronautics and Astronautics, Beijing, China
• 2010 "Enigma of long-non-coding RNA in Vertebrate Biology" - The Chinese University of Hong Kong, Hong Kong, China
• 2010 "Developmental vasculogenesis mechanisms during embryogenesis and disease in vertebrates" - Department of Biology and Center for Chinese Medicine, Hong Kong University of Science and Technology, Clear Water Bay Road, Hong Kong, China
• 2010 "Dusp-5: a putative target for vascular disease" - Caliper Life Sciences, San Francisco, CA
• 2010 "A Robo: Sox nexus in vascular development" - Children's Environmental Health Sciences Core Center, External Advisory Committee Meeting, Intercontinental Hotel, Milwaukee, WI
• 2010 "Vasculogenesis mechanisms in a developing vertebrate" - 2010 Mayo Angiogenesis Symposia, Mackinac Island, MI
• 2011 "Tools for a Successful Career in Science" - K.J. Somaiya College of Science and Commerce, Somaiya Vidyavihar Complex, Vidyavihar-East, Mumbai, India
• 2011 "Vasculogenesis mechanisms during vertebrate embryonic development" - AU-KBC Research Centre, Anna University, Chennai, India

• 2012 "Serendipitous discovery implicates signaling molecules in vascular anomalies" and "Transcriptional Regulation of Roundabout4 (Robo4) in the Developing Vasculature" - Georgia Health Sciences University, Augusta, GA

 Publications (2008-present)

• Garnaas MK, Liu M, Marx R, Li K, Baraban J, Horowitz A, Ramchandran R. Syx, a novel Rho A guanine exchange factor, is essential for angiogenesis in vivo 2008. Circulation Research 2008 Sep 26;103(7):710-6; PMID:  18757825.
• Chun CZ, Kaur S, Samant GV, Wang L, Pramanik K, Garnaas M, Li K, Field L, Mukhopadhyay D, and Ramchandran R. Snrk-1 is involved in multiple steps of angioblast development and acts via notch signaling pathway in artery-vein specification in vertebrates.  Blood 2009, 113(5):1192-1199; PMID:  18723694. **Inside Blood commentary on both articles.
• Pramanik K, Chun CZ, Garnaas MK, Samant GV, Li K, Horswill MA, North PE, and Ramchandran R. Dusp-5 and Snrk-1 coordinately function during vascular development and disease.  Blood 2009, 113(5):1184-1191; PMID:  18927432.  **Inside Blood commentary on both articles.
• Kaur S*, Samant GV*, Pramanik K, Loscombe PW, Pendrak ML, Roberts DD, and Ramchandran R. Silencing of directional migration in robo4 knockdown endothelial cells.  BMC Cell Biology 2008 Nov 39:61; PMID:  18980679.
• Makky K, Duvnjak P, Pramanik K, Ramchandran R and Mayer AN. A whole-animal microplate assay for metabolic rate using zebrafish. Journal of Bio Molecular Screening 2008 Dec; 13(10):960-7; PMID:  19029015.
• Jia Y, Wu S-L, Isenberg JS, Sipes JM, Field L, Zeng B, Bandle RW, Ramchandran R, Karger BL, and Roberts DD.  Thiolutin inhibits endothelial cell adhesion by perturbing Hsp27 interactions with the actin cytoskeleton.  Cell Stress Chaperones 2010 Mar; 15(20:165-81; PMID:  19579057.
• Kanungo J, Zhen, Y, Amin N, Kaur S, Ramchandran R, and Pant, H.  Specific inhibition of cyclin-dependent kinase 5 activity induces motor neuron development in vivo 2009.  Biochemical and Biophysical Research Communications 386 (2009):263-267; PMID:  19523926.
• Li K, Blum Y, Verma A, Liu Z, Pramanik K, Leigh N, Chun C, Samant G, Zhao B, Garnaas M, Horswill M, Stanhope S, North P, Miao R, Wilkinson G, Affolter M, and Ramchandran R.  A noncoding antisense RNA at the tie-1 locus regulates tie-1 transcript levels function in vivo. Blood 2010 Jan7;115(1):133-9; PMID:  19880500.
• Verma A, Bhattacharya R, Remadevi I, Li K, Pramanik K, Samant G, Horswill M, Chun C, Mukhopadhyay D, Ramchandran R, and Wilkinson G.  Ecscr promotes angioblast migration during vasculogenesis and enhances kdr sensitivity.  Blood, 2010 Jun 3; 115(22):4614-22; PMID:  20086248.  **Inside Blood commentary on EC-specific chemotaxis receptor:  a double-edged sword, p.4614.
• Zhao B, Chun C, Liu Z, Horswill M, Pramanik K, Wilkinson G, Ramchandran R, and Miao R. Nogo-B receptor is essential for angiogenesis in vivo via Akt pathway. Blood, 2010. Dec 9;116(24):5423-33. PMID: 20813898.
• Chun CZ, Remadevi I, Schupp M, Samant GV, Pramanik K, Wilkinson GA and Ramchandran R. Fli+ etsrp+ hemato-vascular progenitor cells proliferate at the lateral plate mesoderm during vasculogenesis in zebrafish. PLoS One. 2011 Feb 25;6(2):e14732; PMC3045372.
• Lakshmikanthan S, Sobczak M, Chun CZ, Henschel A, Dargatz J, Ramchandran R, Chrzanowska-Wodnicka M. Rap1b regulates angiogenesis by an endothelial cell-autonomous mechanism involving integrin ávâ3-dependent VEGFR2 activation. Blood 2011. In press. PMID not available.
• Samant GV, Schupp MO, François M, Moleri S, Kothinti RK, Chun CZ, Sinha I, Sellars S, Leigh N, Pramanik K, Horswill MA, Remadevi I, Li K, Wilkinson GA, Tabatabai NM, Beltrame M, Koopman P, and Ramchandran R. Sox factors transcriptionally regulate robo4 expression in the developing vasculature in zebrafish. J. Biol. Chemistry 2011. PMID: 21730073; PMC3162435.
• Patra C, Kim J, Pramanik K, d'Uscio LV, Patra S, Ramchandran R, Strano M, and Mukhopadhyay D. Reactive Oxygen Species Driven Angiogenesis by Inorganic Nanorods. PMID 21967244.
  

1R01HL090712-01A2 (Ramchandran, R.) 2009-2014 - NIH/NHLBI. Roundabout4 signaling in endothelial cells – Role:  PI

1R01HL108938-01 (Miao, R. and Ramchandran, R.) 2011-2016 NIH Role of NgBR - Role: Co-I

1R43NS067749-01 (Ferrante, A and Ramchandran, R) 2010-2012 High content discovery assay for muscular dystrophy therapeutics - Role:  Co-I

HL007792 (Harder, D) 2009-2014 NIH T32 Training Program - Hypertension and Vascular Biology Training Grant

HL007792 (Harder, D) 2009-2014 -  NIH T32 Training Program. Hypertension and Vascular Biology Training Grant Role:  Mentor

(Ramchandran, R.) 2011 - Marjorie Siebert Aylen Foundation.  MYCN Zebrafish Model for Neuroblastoma-  Role:  PI        

  
(Ramchandran, R.) 2011 - Alvin and Marion Birnschein Foundation . MYCN Zebrafish Model for Neuroblastoma-  Role:  PI  

        
(Ramchandran, R.) 2011 - The Donald & Judith Alstadt Charitable Research Foundation- Role:  PI