Developmental Vascular Biology Program

Current events/important calendars

2010 schedule of presenters for Developmental Vascular Biology Program

Vascular Biology Interest Group

Leadership

Ramani Ramchandran, PhD, Director                                                       

 George Wilkinson, PhD, Assistant Professor

Contact information:
8701 Watertown Plank Road
Milwaukee, WI 53226

Dr. Ramchandran phone and e-mail: (414) 955-2387, rramchan@mcw.edu   

Dr. Wilkinson phone and e-mail: (414) 955-2390, gwilkins@mcw.edu

Staff

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 vasculogenesis problem during embryonic vascular development.
Mark Horswill, MS, research associate I - My research philosophy is to understand general ideas and mechanisms fundamental to biological processes in the hope that basic knowledge acquired will have an impact on people's health. Currently, my research interest is to determine the signaling molecules and pathways responsible for artery/vein specification in the developing vasculature by examining structure/functions studies of Snrk-1.
	Noah Leigh, graduate student- My research interests are focused on drug discovery related to the process of angiogenesis.  Currently I am working on the characterization the expression and function of a novel matrix metalloprotease-like gene in embryonic zebrafish development.
Keguo Li, PhD, post-doctoral fellow - I work on the roles of antisense transcripts and the positional cloning of subtelomeric genes. The antisense transcripts arise from the antisense strand of DNA and regulate the translational output of their complementary transcripts. Subtelomeric sequences vary from individual to individual and are underrepresented in conventional genomic libraries, therefore, an alternative method needs to be developed to circumvent the drawbacks of chromosome walking.
	Kallal Pramanik, PhD, post-doctoral fellow - My research involves identifying targets for vascular related diseases. Vessel formation involves two fundamentally distinct processes, vasculogenesis, the process of differentiation of precursor cells into endothelial cells, and angiogenesis, the process of endothelial sprouting from established vasculature. I am interested in studying the regulators that play important roles in maintaining and migration of the endothelial precursor cells during vasculogenesis.
Ganesh Samant, PhD, post-doctoral fellow - My broad research interests include investigation of the bio-molecular pathways that underlie malignancy and metastasis. Specifically, I want to elucidate intracellular mechanisms that lead to pathological angiogenesis. I utilize molecular biology techniques coupled with pharmacological functional assays to answer problems associated with pathological and physiological angiogenesis.
	Indranil Sinha, PhD, post-doctoral fellow - In a developing embryo, in addition to blood vessels, a second vessel system, namely lymphatics, is necessary to clear toxins and other deleterious substances and molecules from tissue spaces. My research investigates genes that are important for the development of the lymphatic vessels. Lymphatic endothelial cells (LECs) line the lymphatic vessels and they arise from venous endothelial cells (VECs). Prox1, a DNA binding protein is responsible for maintaining the identity of LECs and turning off Prox1 switches LECs into VECs. If successful, my studies will shed light on the mechanisms that may have gone wrong in patients with defective lymphatic function leading to accumulation of fluid under the skin and will benefit patients with lymphedema.

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.
Anjali Verma, PhD, post-doctoral fellow - My specific interest lies in understanding the molecular mechanisms associated with early vascular development. I am using zebrafish as a model system to study embryonic vascular development.

Research in the Developmental Vascular Biology Program

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 axon guidance. We study one member of the axon guidance family, roundabout4 (robo4), and its role in this process.
• Translational disease models in fish
  Vessel growth is tightly regulated during development. Any imbalance in this regulation often is associated with disease. For example, tumor growth is dependant 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.

Wilkinson lab:

• Role of vascular EphrinB2 in lung development
  To understand lung alveolar development, the Wilkinson Lab is undertaking a molecular genetics approach using knockin mice. In particular, the laboratory studies mice with a mutation in the gene ephrinB2, which is expressed in the vasculature. Mice homozygous for this mutation are born with normal lung appearance, but subsequently show profound postnatal lung defects, including an almost complete failure to form alveoli. The abnormal lung development of these mutant mice provides an opportunity to investigate the phenomenon of alveolar initiation from a cell and developmental biology point of view. We would like to know how the loss of function of ephrinB2 causes failed alveolar development. We also hope to learn which vasculature and what signals contribute to the process.

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

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.

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 "Functional characterization of Snrk-1 gene in developing vasculature – an update". CVRC Works-in-Progress, Medical College of Wisconsin, Milwaukee, WI
• 2010 "Vasculogenesis mechanisms in vertebrates". CBNA Presentation, Medical College of Wisconsin, Milwaukee, WI
• 2010 "How to make sense of anti-sense RNA". CRI Noon Conference, Medical College of Wisconsin, Milwaukee, WI
• 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

 Publications (2007-present)

• Kaur S, Castellone MD, Bedell VM, Konar M, Gutkind JS, and Ramchandran R.  Robo4 signaling in endothelial cells imply attraction guidance mechanisms.  J. Biol. Chemistry 2006, 281(16):11347-56.
• Bedell V, Yeo SY, Park KW, Chung J, Seth P, Shivalingappa V, Zhao J, Obara T, Sukhatme VP, Drummond I, Li DY, and Ramchandran R.  Roundabout4 is essential for angiogenesis in vivo.  PNAS 2005, 1023:6373-78.
• Arbiser JL, Kau T, Konar M, Narra K, Ramchandran R, Summers SA, Vlahos CJ, Ye K, Perry BN, Matter W, Fischl A, Coolk J, Silver PA, Bain J, Cohen P, Whitmire D, Furness S, Govindarajan B, and Bowen P. Solenopsin, the alkaloidal component of the fire ant (Solenopsis invicta), is a naturally occurring inhibitor of phosphatidylinositol-3-kinase signaling and angiogenesis. Blood 2007, 109 (2):560-5.
• Kanungo J, Li BS, Goswami M, Zheng YL, Ramchandran R and Pant HC. Cloning and characterization of zebrafish (Danio rerio) Cyclin dependent kinase 5. Neurosci Lett. 2007, 412 (3): 233-8.
• Isenberg JS, Jia Y, Field L, Ridnour LA, Sparatore A, Soldato PD, Sowers AL, Mitchell JB, Yeh GC, Moody TW, Wink DD, Ramchandran R, and Roberts, DD. Modulation of Angiogenesis by Dithiolethione-modified NSAIDs and Valproic Acid. Br. J. Pharmacol. 2007, 151 (1): 63-72.
• Kaur S, Abu-Abab MS, Singla S, Yeo SY, and Ramchandran R. Expression pattern for unc5b, an axon guidance gene in embryonic zebrafish development. Gene Expression 2007, 13 (6): 321-7.
• 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.
• 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. **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.  **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.
• 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.
• 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 and Chaperones, In press.
• 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.

• 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 First Edition Paper, prepublished online October 30, 2009; DOI 10.1182/blood-2009-09-242180.

 Press

• 1997 - Interview and picture appeared in the MCG Today magazine regarding graduate students' perspective on current issues such as NIH funding.
• October 15, 1999 - Organization brings together Indian scientists.  Bulletin in Focus (Harvard Medical School publication)
• Summer 1999 - Intent on killing cancer - Scientists move closer to a new approach.  Basic Research in Pulse (Beth israel Deaconess Medical Center)
• May 4, 2001 - First cell surface receptor identified for endostatin.  Research brief in Focus (Harvard Medical School publication)
• August 2006 - PNAS paper highlighted in the CCR newsletter, Frontiers in Science http://ccr.cancer.gov/news/frontiers/august2006/ramchandran.asp
• March 2007 – Our research program has been highlighted in the "Searchlight" e-newsletter (Children's Research Institute, Medical College of Wisconsin)
• March 2007 – Faculty appointment – Press release – Medical College of Wisconsin
• May 2007 – Investigation of Blood Vessel Formation Strives to Understand Cancer Tumor Growth – Article in MCW Cancer Center News (Medical College of Wisconsin)
• October 2007 - State Tax Check-Off Program Raises $115,000 for MCW Breast Cancer Research – Ganesh Samant, post-doctoral fellow award announcement
• 2008 - Kaur S*, Samant GV*, Pramanik K, Loscombe PW, Pendrak ML, Roberts DD and Ramchandran R. Silencing of directional migration in robo4 knockdown endothelial cells 2008. BMC Cell Biology in press.  BMC Cell Biology Article figure chosen as "image of month" for publication online.  http://www.biomedcentral.com/bmccellbiol/imageofthemonth/2008/12  BMC Cell Biology Article:  Highly Accessed Article Citation given based on number of downloads in a given month for this article 
• January 29, 2009 – Press coverage concerning article - Dusp-5 and Snrk-1 coordinately function during vascular development and disease 2008.  Blood 2009, 113 (5):1184-1191.   Birthmark or Blood Vessel Problem?
  • The Washington Post http://www.washingtonpost.com/wp-dyn/content/article/2009/01/29/AR2009012903270.
  • News Wise http://www.newswise.com/articles/view/548143/.
  • MSN Health and Fitness http://health.msn.com/kids-health/articlepage.aspx?cp-documentid=100232148.
  • Forbes.com http://www.forbes.com/feeds/hscout/2009/01/29/hscout623304.
  • Yahoo News  http://news.yahoo.com/s/hsn/20090130/hl_hsn/birthmarkorbloodvesselproblem.
  • Womenshealth.gov (The federal government source for women's health)  http://www.womenshealth.gov/news/english/623304.
  • The Investigator (Children's Research Institute newsletter) http://chw.org/vascularbiology
• April 2009, Medical College of Wisconsin Receives Grant to Study Vascular Tumor Proteins, MCW Press Release             http://infoscope.mcw.edu/NewsCenter/CollegeNews/Collegereceivesgranttostudyvasculartumorproteins.htm

Current funding

1K22CA095325-01 (Ramchandran, R.) 2002-2009 - NIH/NCI Scholar Award. Role:  PI       

AHW 5520106 (Ramchandran, R.) 2008-2011 - Advancing Healthier Wisconsin. Cloche:  A basic and translational model for cardiovascular disease - Role:  PI

1R01HL090712-01A2 (Ramchandran, R.) 2009-2013 - NIH/NHLBI. Roundabout4 signaling in endothelial cells – Role:  PI

3R01HL090712-01A2S1 (Ramchandran, R.) 2009-2011  - NIH/NHLBI. Administrative Supplement for Roundabout4 signaling in endothelial cells – Role:  PI   

Christmas 2009 - Donation to Hunger Task Force