The Stowers Institute For Medical Research
   TrainingPrograms  LecturesPublications
The Stowers Institute for Medical Research home page
Key people and information
A multi-disciplined approach to basic research
Research Campus
Up-to-date news on the Institute and the scientific community
How To Help
Kulesa Lab
Paul Kulesa, Ph.D.
Director of Imaging/Kulesa Lab
pmk@stowers.org
Neural crest cell migration, cancer metastasis, biological imaging

Our Focus
     Cell migration in the vertebrate embryo and cancer metastasis share similar features of cell plasticity and invasive cell behavior, yet it is unclear whether there are common signaling pathways that regulate these processes. A fundamental problem has been our limited ability to study cell movements in an intact model system and couple quantitative information on cell behaviors to underlying molecular mechanisms. We use the chick neural crest and neural crest-derived cancers to address this question.

     We believe that by working at the interface of a multipotent stem-cell like population such as the neural crest and its ancestrally related melanoma, we will discover how molecular signals choreograph the embryonic cell invasion program and whether these signals can reprogram metastatic cancer to a less destructive phenotype.

The Neural Crest and Melanoma
      The neural crest are a highly invasive, multipotent cell population with derivatives that contribute to the face, heart, and trunk peripheral nervous system. Neural crest cells arise all along the vertebrate axis and sort into discrete migratory pathways that reach precise targets. Neural crest-related birth defects often entail a failure of cell navigation, however the cellular and molecular mechanisms that program the neural crest migratory pattern are still unclear. In contrast to the embryo, neural crest-derived cancer cell types, including melanoma and neuroblastoma are highly aggressive and display phenotypic plasticity. Our hypothesis is that there is a convergence of embryonic and tumorigenic signaling mechanisms that regulate cell plasticity and cell invasion.

Our Approach
     We develop innovative cell labeling and live cell and embryo imaging to follow single cell behaviors. We use molecular biology, tissue transplantation, and embryo microsurgery to perturb normal cell behaviors. We use microinjection, targeted electroporation and photoactivation to selectively label cells. We extract quantitative information from 3D/4D biological image sequences.

Projects:

1. An In Vivo Analysis of the Mechanisms of Neural Crest Migration

      Disruption of a neural crest cell’s ability to interpret environmental signals lead to migratory patterning defects. We investigate the role of the embryonic microenvironment in signaling and modulating neural crest migration, with particular focus on neuropilin/VEGF interactions. We have shown that chemotactic and cell communication signaling is critical to cell invasion of the branchial arches and are following up on how specific receptor-ligand interactions regulate this behavior.

      In the trunk, we are interested in sympathetic ganglia formation and have shown that complex cell behaviors shape cells into discrete cell clusters. We have discovered that eph/ephrin and N-cadherin signaling regulate the pattern of cell clusters and chemokine signaling attracts cells to the dorsal aorta.

2. The Embryonic Neural Crest Microenvironment as a Model for Cancer Cell Reprogramming

     The chick embryonic neural crest-rich microenvironment provides an attractive model system to explore tumor cell reprogramming and metastatic ability. We have shown that human metastatic melanoma cells transplanted into the embryonic chick neural tube migrate to host neural crest cell target sites, do not form tumors, and a subset of these tumor cells are reprogrammed to a neural crest cell-like phenotype. We are currently investigating the functional role of key molecular mechanisms that modulate neural crest migration to determine the relevance of these signaling pathways involved in the control of tumor cell fate determination and reprogramming of the metastatic phenotype.

3. Development of Innovative Tools for Tracing Cell Movements in Embryos

     Cells interpret and integrate signals from their local microenvironment, yet our ability to connect cell signaling events with cell behaviors are limited. We develop novel techniques to fluorescently mark and follow cells, using photoactivation and multispectral imaging, and interface cells and embryos with long term time-lapse microscopy, using novel microfluidics and microfabrication designs for cell, tissue slice, and embryo culture.

Academic Appointment: Associate Professor, Department of Anatomy & Cell Biology, The University of Kansas School of Medicine

Selected publications

Kulesa PM, Stark DA, Steen J, Lansford R, Kasemeier-Kulesa JC. Watching the assembly of an organ a single cell at a time using confocal multi-position photoactivation and multi-time acquisition. Organogenesis. 2009;5:238-247.

McLennan R, Teddy JM, Kasemeier-Kulesa JC, Romine MH, Kulesa PM. Vascular Endothelial Growth Factor Regulates Cranial Neural Crest Migration In Vivo [published ahead of print December 24 2009]. Dev Biol. 2009. Abstract

Kulesa PM, Lefcort F, Kasemeier-Kulesa JC. The migration of autonomic precursor cells in the embryo [published ahead of print September 29 2009]. Auton Neurosci. 2009. Abstract

Kasemeier-Kulesa JC TJ, Postovit LM, Seftor EA, Seftor RE, Hendrix MJ, Kulesa PM. Reprogramming Multipotent Tumor Cells with the Embryonic Neural Crest Microenvironment. Dev Dyn. 2008.

Kulesa PM, Teddy JM, Stark DA, Smith SE, McLennan R. Neural Crest Invasion is Spatially-Ordered Progression Into the Head with Higher Cell Proliferation at the Migratory Front as Revewaled by the Photactivatable Protein, KikGR. Dev Biol. 2008.

Rupp PA, Kulesa PM. A role for RhoA in the two-phase migratory pattern of post-otic neural crest cells. Dev Biol. 2007. Abstract

Kulesa PM, Schnell S, Rudloff S, Baker RE, Maini PK. From segment to somite: Segmentation to epithelialization analyzed within quantitative frameworks. Dev Dyn. 2007;236:1392-1402. Abstract

Stark DA, Kulesa PM. An in vivo comparison of photoactivatable fluorescent proteins in an avian embryo model. Dev Dyn. 2007. Abstract

Hendrix MJC, Seftor EA, Seftor REB, Kasemeier-Kulesa J, Kulesa PM, Postovit L-M. Reprogramming metastatic tumor cells with embryonic microenvironments. Nat Rev Cancer. 2007;7:246-255.

McLennan R, Kulesa PM. In vivo analysis reveals a critical role for neuropilin-1 in cranial neural crest cell migration in chick. Dev Biol. 2006

Kasemeier-Kulesa JC, Bradley R, Pasquale EB, Lefcort F, Kulesa PM. Eph/ephrins and N-cadherin coordinate to control the pattern of sympathetic ganglia. Development. 2006;133:4839-4847. Abstract

Kulesa PM, Kasemeier-Kulesa JC, Teddy JM, Margaryan NV, Seftor EA, Seftor RE, Hendrix MJ. Reprogramming metastatic melanoma cells to assume a neural crest cell-like phenotype in an embryonic microenvironment. Proc Natl Acad Sci U S A. 2006;103:3752-3757. Abstract

Kulesa PM, Lu CC, Fraser SE. Time-Lapse Analysis Reveals a Series of Events by Which Cranial Neural Crest Cells Reroute around Physical Barriers. Brain Behav Evol. 2005;66:255-265. Abstract

Stark D, Kulesa PM. In vivo marking of single cells in chick embryos using photoactivation of GFP. Current Protocols in Cell Biology. 2005; Supplement 28:12.18.11-12.18.11.

Teddy JM, Lansford R, Kulesa PM. Four-Color, 4D Time-Lapse Confocal Imaging of Chick Embryos. Biotechniques. 2005;39:703-710.

Stark DA, Kulesa PM. Photoactivatable green fluorescent protein as a single-cell marker in living embryos. Dev Dyn. 2005;233:983-992. Abstract

Kasemeier-Kulesa JC, Kulesa PM, Lefcort F. Imaging neural crest cell dynamics during formation of dorsal root ganglia and sympathetic ganglia. Development. 2005;132:235-245. Abstract

Teddy JM, Kulesa PM.  In vivo evidence for short- and long-range cell communication in cranial neural crest cells. Development. 2004 Dec;131(24):6141-51. Abstract

Kulesa PM. Developmental imaging: Insights into the avian embryo. Birth Defects Res C Embryo Today. 2004 Sep;72(3):260-6. Abstract

Kulesa PM, Fraser SE. In Ovo Imaging of Avian Embryogenesis. In: R Yuste, and A Konnerth, eds. Imaging in Neuroscience and Development: A Laboratory Manual. New York: Oxford: Cold Spring Harbor Laboratory Press; Lavis Marketing; 2004:700 p.

Kasemeier J, Lefcort F, Fraser SE, Kulesa PM. A novel sagittal slice explant technique for time-lapse imaging of the formation of the chick periperal nervous system. In: R Yuste, and A Konnerth, eds. Imaging in neuroscience and development : a laboratory manual. R. Yuste and A. Konnerth ed. New York: Oxford: Cold Spring Harbor Laboratory Press; Lavis Marketing; 2004:700 p.

Kulesa PM, Ellies DL, Trainor PA. Comparative analysis of neural crest cell death, migration, and function during vertebrate embryogenesis. 2004; Dev Dyn. 229:14-29. Abstract.

Patten I, Kulesa PM, Shen MM, Fraser SE, Placzek M. . Distinct modes of floor plate induction in the chick embryo. Development. 2003;130:4809-4821. Abstract.

Kulesa PM, Fraser SE.  Cell dynamics during somite boundary formation revealed by time-lapse analysis. Science. 2002;298:991-995. Movies.

Comments about this paper may be found in Developmental Cell, 3, 605-613, The Scientist, 17, 2 (10), and Caltech News, 12 Nov.

Jones EAV, Crotty D, Kulesa PM, Waters CW, Baron MH, Fraser SE, Dickinson ME.  Dynamic in-vivo imaging of post-implantation mammalian embryos using whole embryo culture. Genesis. 2002;34:228-235.

Mathis L, Kulesa PM, Fraser SE.  FGF receptor signaling is required for the maintenance of neural progenitors during Hensen's node progression. Nat Cell Biol. 2001;3:559-566. Comments about this paper may be found in Nature Reviews Neuroscience 2, 381, Nature Cell Biology, 3 June, Cell 106, 133-136, and Caltech News, 21 June.

Kulesa PM, Bronner-Fraser M, Fraser SE. In ovo time-lapse analysis after dorsal neural tube ablation shows rerouting of chick hindbrain neural crest. Development. 2000;127: 2843-2852.

Kulesa PM, Fraser SE.  In ovo time-lapse analysis of chick hindbrain neural crest cell migration shows interactions during migration to the branchial arches. Development. 2000;127:1161-1172. Movies.

Kulesa PM, Fraser SE. Neural Crest Cell Dynamics Revealed By Time-Lapse Video Microscopy Of Whole Embryo Chick Explant Cultures. Dev Biol. 1998;204:327-344. Comments about this paper may be found in Science 288, 7 April.

Kulesa PM, Fraser SE.  Confocal imaging of living cells in intact embryos. Methods Mol Biol. 1998;122:205-222. Abstract.

Kulesa PM, Fraser SE. Segmentation of the vertebrate hindbrain: a time-lapse analysis. Intl J Dev Biol. 1998;42:385-392.

Krull CE,  Kulesa PM . Embryonic Explant and Slice Preparations for Studies of Cell Migration and Axon Guidance . In: de Pablo F, Ferrus A, Stern C, eds. 36. Cellular Techniques in Developmental Biology. New York: Academic Press;1998.

Burgess PK, Kulesa PM, Murray JD,  Alvord EC Jr.  The Interaction of Growth Rates and Diffusion Coefficients in a Three-dimensional Mathematical Model of Gliomas. J Neuropath  Exp Neurol. 1997;56:704-713

Kulesa PM,  Cruywagen GC, Lubkin SR,  Maini PK, Sneyd J, Ferguson MWJ, Murray JD. On A Model Mechanism for the Spatial Patterning of Teeth Primordia in the Alligator. J Theoret Biol. 1996;180:287-296.

Murray JD, Kulesa PM. On a dynamic reaction-diffusion mechanism for the spatial patterning of teeth primordia in the alligator. J Chem Soc Faraday Trans. 1996;92:2927-2932.

Return to Research Team