Novel Model Systems for Discovering New Therapeutic Targets

1:00 Prof. Dennis CleggResearch in Stem Cell Biology and Engineering

1:20 Harriman Prof. Kenneth KosikMicroRNAs: a Novel Class of Targets for Medical Intervention

1:40- Break

1:50 Prof. Joel RothmanAging and Age-related Research

2:10 Prof. Thomas WeimbsNew Functions of Polycystin-1; Polycystic Kidney Disease

2:30 Prof. Michael MahanVaccines against Emerging Infectious Agents

Dennis Clegg

Professor and Chair,Department of Molecular,

Cellular & DevelopmentalBiology

Professor, BiomolecularSciences & EngineeringGraduate Program

Stem Cell Research

Interdisciplinary Research in Stem Cell Biology and

Engineering

Dennis O. CleggProfessor and Chair

Department of Molecular, Cellular and Developmental BiologyNeuroscience Research Institute

University of CaliforniaSanta Barbara

Michael May: Blindness cured by corneal stem cell implant + corneal transplant

Copyright 2003, American Association for the Advancement of Science

Test new drugs on tissues generated from stem cells in vitro.

Faster drug development.

What is the genetic program that controls cell differentiation?

Faster gain in knowledge.

www.stemcells.nih.gov

UCSB Consortium for Stem Cell Biologyand Engineering

Interdisciplinary collaboration between biologists, engineers, ethicists, and neuroscientists representing 18 different departments and centers

• Don Anderson, Director, Center for the Study of Macular Degeneration, Neuroscience Research Institute• Andrew Cleland, Professor, Department of Physics

Dennis O. Clegg, Professor and Chair, Department of Molecular, Cellular and Developmental Biology (also NRI, CSMD, BMSE)• Francis Doyle, Professor, Department of Chemical Engineering (BMSE)• Patrick Daugherty, Assistant Professor, Department of Chemical Engineering (BMSE)• Stuart Feinstein, Co-Director, Neuroscience Research Institute; Professor, Department of Molecular, Cellular and Developmental Biology• Steve Fisher, Professor, Department of Molecular, Cellular and Developmental Biology (NRI)• Kathy Foltz, Associate Professor, Department of Molecular, Cellular and Developmental Biology• Michael Gazzaniga, Professor, Department of Psychology, Director, Sage Center for the Study of the Mind.• Anita Guerrini, Professor, Department of Environmental Studies (History)• Craig Hawker, Professor, Department of Chemistry and Biochemistry; Director, MRL• Evelyn Hu, Professor, ECE and Materials, Co-Director, California NanoSystems Institute.• Lincoln Johnson, Associate Director, Neuroscience Research Institute, Associate Director, Center for the Study of Macular Degeneration, Adjunct Professor, MCDB• Lois Jovanovic, Director, Sansum Medical Research Institute, Adjunct Professor, BMSE• Mustafa Khammash, Director, Center of Control, Dynamical Systems and Computations; Professor, Department of Mechanical and Environmental Engineering• Tod Kippen, Assistant Professor, Department of Psychology (NRI)• Ken Kosik, Co-Director, Neuroscience Research Institute; Professor, Department of Molecular, Cellular and Developmental Biology (BMSE)• David Low, Professor and Vice Chair, Department of Molecular, Cellular and Developmental Biology (BMSE)• B. S. Manjunath, Professor, Department of Electrical and Computer Engineering• Carl Meinhart, Associate Professor, Department of Mechanical and Environmental Engineering • Daniel Morse, Professor, Department of Molecular, Cellular and Developmental Biology; Director, Institute for Collaborative Biotechnologies (BMSE)• James Murai, Research Scientist, Advanced Cell Technology, Inc.• Michael Osborne, Associate Professor of Environmental Studies (History)• Linda Petzold, Professor and Chair, Department of Computer Science (MEE)• Norbert Reich, Professor, Department of Chemistry and Biochemistry (BMSE) • Joel Rothman, Professor, Department of Molecular, Cellular and Developmental Biology (NRI, BMSE)• Erkki Ruoslahti, Distinguished Professor, Department of Molecular, Cellular and Developmental Biology, Distinguished Professor, Burnham Institute for Medical Research. • Cyrus Safinya, Professor, Department of Physics (Materials, MCDB, BMSE)• Geoff Sargent, Research Scientist, Advanced Cell Technology, Inc.• Ambuj Singh, Professor, Department of Computer Science• William Smith, Professor, Department of Molecular, Cellular and Developmental Biology (NRI, BMSE) • Evan Snyder, Professor, Burnham Institute, Director, Stem Cells and Regeneration, Burnham Institute• Hyongsok Tom Soh, Assistant Professor, Department of Mechanical and Environmental Engineering (BMSE) • James Thomson, Scientific Director, Wicell Institute; Professor, Department of Anatomy, University of Wisconsin• Matthew Tirrell, Dean, College of Engineering, The Richard A. Auhll Professor, Department of Chemical Engineering, (also Materials)• Michael West, President and Chief Scientific Officer, Advanced Cell Technology, Inc.

UCSB Consortium for Stem Cell Biology and Engineering

Research Goals:• Understand the basic biology of embryonic and

adult stem cell differentiation.

• Discover new methods for stem cell growth, sorting and delivery.

• Focus on generation of ocular cells for treatment of eye disease.

• November 2, 2004: California passes Proposition 71, creating the California Institute for Regenerative Medicine, funding 3 billion dollars over 10 years. Bond issue blocked by lawsuits.Other states have passed initiatives with lower levels of funding.

• April 10, 2006: CIRM awards training grants using philanthropic funding; UCSB receives 1.3M for Stem Cell Training Program.

• April 21, 2006: Alameda court upholds constitutionality of stem cell program in its entirety. Opponents vow to appeal…

Stem Cell BiologyAdult Stem Cells

Mechanism and functional consequences of neural stem cell differentiation in the adult mammalian brain.– Todd Kippen

Stem cell activation during retinal degeneration.– Steve Fisher

Embryonic Stem CellsRole of Micro RNAs in regulating differentiation of human embryonic stem cells (hES).– Ken Kosik

Genes specifying stem cell competence states. – Joel Rothman / James Thomson

Growth factors that regulate stem cell fate. – William Smith / James Thomson

Stem Cell EngineeringStem Cell surface epitope mapping and screening using peptide libraries.

– Patrick Daugherty

Synthetic and natural extracellular matrix regulation of stem cell fate. – Dennis Clegg, Craig Hawker, Matt Tirrell

Development of new cell sorting technology. – Tom Soh / James Thomson www.lifecell.com

DACS –DielectrophoresisActivated Cell Sorting

www.foresight.org

Copyright 2005, National Academy of Science

Stem Cells and Ocular BiologyDifferentiation of hES to generate Retinal Pigment Epithelial (RPE) cells.- Sherry Hikita / Linc Johnson / Irina Klimanskaya / Robert Lanza / Ken Kosik / Mike West / Dennis Clegg

From The Human Brain Coloring Book,Copyright 1985,Coloring Concepts, Inc.National Eye Institute

www.csmd.ucsb.eduwww.csmd.ucsb.edu

Age-Related Macular Degeneration

• AMD is the leading cause of blindness in people over the age of 55 in North America.

• The disease is characterized by progressive loss of fine acuity vision in central portion of the visual field.

• Loss of vision is due to death of Retinal Pigment Epithelium (RPE) and photoreceptors in macula, possibly due to autoimmune attack,loss of attachment, or accumulation of toxic byproducts.

• Candidate disease for cell-based therapeutics.

The Macula

The macula is a cone rich, circular area of the central retina, about 1/10 inch in diameter,

located next to the optic nerve.

Macula

AHAF.org

Copyright 2005, Cloning and Stem Cells, Mary Ann Leibert Publishers, Inc.

hES-Derived RPE Express RPE Markers

Red = AutofluorescentLipofuscin

Green = Emmprin

hES-Derived RPE ultrastructure is similar to native RPE

hES-RPE retain expression of melanin granules after explantation but not after cell

dissociation with trypsin

Original Culture Explanted Colony Trypsinized Colony

RPE Identity: MultiRPE Identity: Multi--step Commitmentstep Commitment

To identify miRNA ‘expression signatures’ in developmental stages of retinal cell differentiation:

Oct 4 +ESC

Oct 4 +ESC

Oct 4 -

Pax6 -

Pigment -

Pax6 +

Pigment +

Mitf - Mitf +

Undifferentiated hESC

Differentiated hESC

Retinal Pigment Epithelium Precursor

Retinal Precursor

Retinal Pigment Epithelium

Six3Rx

Otx1/Otx2

•Compare with miRNA profile of fhRPE

•Assay for RPE65 (mRNA, protein)

Six3: 4

Rx: 3

Pax6: 9

Otx1: 27

Otx2: 11

Mitf: 20

Number of miRNAs Predicted to Target Key Transcription Factors in

Retinal Development

Conclusions:1. Stem cells can be coaxed to differentiate into ocular

cells, including retinal pigment epithelium-like cells.

2. RPE derived from hES display many characteristics of native RPE.

3. Future research will address regulation of differentiation, functional abilities, and stability of phenotype.

AcknowledgementsUCSB• Sherry Hikita• Erin Dunkle• Dave Buchholz• Sergiu Leu• Julian Davis• Vivian Ericson• Steve Morris• Kathryn Blaschke• Andy Reese

• Linc Johnson• Don Anderson• Christy Curletti• Monte Radeke• Pat Johnson• Ken Kosik

• Craig Hawker• Joel Rothman

Collaborators• Mike West, Robert Lanza, Irina Klimanskaya,

Geoff Sargent, Jim Murai, Advanced Cell Technology

• Babak Azad, CDNA Microarray Inc• Igal Gery, National Eye Institute• David Jackson, Genentech• Jun Parsons, Evan Snyder, Burnham Institute

Funding: California Institute for RegenerativeMedicine, NIH, California Tobacco-RelatedDisease Research Program, Fight for Sight,American Health Assistance Foundation,Cottage Hospital Research Program, TriCounties Blood Bank Fellowship, Sigma Xi