Engineering of the Extracellular Matrix: Working toward Neural Stem Cell Programming and Neurorestoration— Concept and Progress Report

IN THE CONCEPT of neurorestoration, cellular and structural elements that have been lost are replaced, and their function is restored. Central to this therapeutic strategy is the transplantation of neural progenitor cells such as clonogenically expanded stem cells. Stem cells make decisions regarding fate and patterning in response to external environmental signals. The therapeutic effectiveness of neural stem cells may be facilitated by the ability to manipulate these signals in a temporal and spatially appropriate fashion. Artificial deoxyribonucleic acid and artificial protein technology combines elements of protein engineering, molecular biology, and recombinant deoxyribonucleic acid technology to produce proteins with functional domains derived from naturally occurring proteins and represents a potentially powerful tool to modulate stem cell behavior. To this end, we have developed three artificial extracellular matrix proteins that incorporate the active domain of hJagged1 and hDelta1 into an elastin backbone. hJagged1 and hDelta1 are members of the DSL family of ligands to the Notch receptor, a signaling system that is very important in development and is the strongest known signal to instruct neural progenitor cells to choose glial fates over neuronal fates. The successful cloning of the artificial genes was confirmed by test digestions with appropriate restriction enzymes as well as direct deoxyribonucleic acid sequencing. In addition, we have demonstrated that all three artificial extracellular matrix proteins express at a high level in a prokaryotic host. This report describes the concept and progress in an entirely novel and previously unreported approach to modulate neural stem cell behavior. Its future application could include in vitro processing of stem cells before transplantation, supporting and programming the cells after transplantation, as well as the development of bioactive biomaterials.

[1]  I. Weissman,et al.  Stem Cells Units of Development, Units of Regeneration, and Units in Evolution , 2000, Cell.

[2]  T. Palmer,et al.  Fibroblast Growth Factor-2 Activates a Latent Neurogenic Program in Neural Stem Cells from Diverse Regions of the Adult CNS , 1999, The Journal of Neuroscience.

[3]  F M Watt,et al.  Out of Eden: stem cells and their niches. , 2000, Science.

[4]  J. García-Verdugo,et al.  Noggin Antagonizes BMP Signaling to Create a Niche for Adult Neurogenesis , 2000, Neuron.

[5]  J Kimble,et al.  lag-2 may encode a signaling ligand for the GLP-1 and LIN-12 receptors of C. elegans. , 1994, Development.

[6]  J. V. Hest,et al.  Protein-based materials, toward a new level of structural control. , 2001, Chemical communications.

[7]  T. Mcintosh,et al.  Fetal hippocampal transplants attenuate CA3 pyramidal cell death resulting from fluid percussion brain injury in the rat. , 1995, Journal of neurotrauma.

[8]  J. Loturco,et al.  Neuronal Differentiation of Precursors in the Neocortical Ventricular Zone Is Triggered by BMP , 1998, The Journal of Neuroscience.

[9]  S. Rafii,et al.  Endothelial Trophic Support of Neuronal Production and Recruitment from the Adult Mammalian Subependyma , 1999, Molecular and Cellular Neuroscience.

[10]  L. Olson,et al.  Growth characteristics of adrenergic nerves in the adult rat. Fluorescence histochemical and 3H-noradrenaline uptake studies using tissue transplantations to the anterior chamber of the eye. , 1970, Acta physiologica Scandinavica. Supplementum.

[11]  I. Greenwald,et al.  Interchangeability of Caenorhabditis elegans DSL proteins and intrinsic signalling activity of their extracellular domains in vivo. , 1995, Development.

[12]  S. Pons,et al.  Vitronectin regulates Sonic hedgehog activity during cerebellum development through CREB phosphorylation. , 2001, Development.

[13]  A. Israël,et al.  Delta-1 Activation of Notch-1 Signaling Results inHES-1 Transactivation , 1998, Molecular and Cellular Biology.

[14]  S. Aamodt.  Young neurons in old brains , 2003, Nature Neuroscience.

[15]  Jonas Frisén,et al.  Identification of a Neural Stem Cell in the Adult Mammalian Central Nervous System , 1999, Cell.

[16]  T. Mcintosh,et al.  Combined fetal neural transplantation and nerve growth factor infusion: effects on neurological outcome following fluid-percussion brain injury in the rat. , 1996, Journal of neurosurgery.

[17]  D. van der Kooy,et al.  Notch pathway molecules are essential for the maintenance, but not the generation, of mammalian neural stem cells. , 2002, Genes & development.

[18]  K. Na,et al.  Incorporation of Sulfonylurea into N-Isopropylacrylamide as an Extracellular Matrix for an Artificial Pancreas , 2001, Bioscience, biotechnology, and biochemistry.

[19]  Eric M. Morrow,et al.  rax, Hes1, and notch1 Promote the Formation of Müller Glia by Postnatal Retinal Progenitor Cells , 2000, Neuron.

[20]  A. Kiger,et al.  Somatic support cells restrict germline stem cell self-renewal and promote differentiation , 2000, Nature.

[21]  Richard P. Haugland,et al.  Handbook of fluorescent probes and research chemicals , 1996 .

[22]  M. Elices,et al.  Extracellular matrix proteins expressed by human adult astrocytes in vivo and in vitro: An astrocyte surface protein containing the CS1 domain contributes to binding of lymphoblasts , 1997, Journal of neuroscience research.

[23]  M. Schwab.  Repairing the Injured Spinal Cord , 2002, Science.

[24]  B. Barres,et al.  Up a Notch Instructing Gliogenesis , 2000, Neuron.

[25]  T. Yamaoka,et al.  Effects of temperature and pressure on the aggregation properties of an engineered elastin model polypeptide in aqueous solution. , 2000, Biomacromolecules.

[26]  M. Mehler.  Mechanisms regulating lineage diversity during mammalian cerebral cortical neurogenesis and gliogenesis. , 2002, Results and problems in cell differentiation.

[27]  L. Olson,et al.  Brain tissue transplanted to the anterior chamber of the eye , 1976, Cell and Tissue Research.

[28]  D. van der Kooy,et al.  Why stem cells? , 2000, Science.

[29]  Ting Xie,et al.  Germline Stem Cells Anchored by Adherens Junctions in the Drosophila Ovary Niches , 2002, Science.

[30]  Monte A. Gates,et al.  Site-Specific Migration and Neuronal Differentiation of Human Neural Progenitor Cells after Transplantation in the Adult Rat Brain , 1999, The Journal of Neuroscience.

[31]  David J. Anderson,et al.  Alternative Neural Crest Cell Fates Are Instructively Promoted by TGFβ Superfamily Members , 1996, Cell.

[32]  J. Flanagan,et al.  Nogo Domains and a Nogo Receptor:Implications for Axon Regeneration , 2001, Neuron.

[33]  D. Tirrell,et al.  Engineering the extracellular matrix: a novel approach to polymeric biomaterials. I. Control of the physical properties of artificial protein matrices designed to support adhesion of vascular endothelial cells. , 2000, Biomacromolecules.

[34]  Thiery Michon,et al.  Les protéines artificielles , 2000 .

[35]  S. Giulini,et al.  Complete in vitro prosthesis endothelialization induced by artificial extracellular matrix. , 1999, Journal of investigative surgery : the official journal of the Academy of Surgical Research.

[36]  M. Wegner,et al.  Bone Morphogenetic Proteins Are Required In Vivo for the Generation of Sympathetic Neurons , 1999, Neuron.

[37]  Jenny E Rooke,et al.  Positive and negative signals between interacting cells for establishing neural fate , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.

[38]  I. Weissman.  Stem cells--scientific, medical, and political issues. , 2002, The New England journal of medicine.

[39]  M. Apuzzo.  Cellular and Molecular Neurosurgery: Fetal Grafting to Treat Parkinson's Disease , 2001 .

[40]  H. Vandenburgh,et al.  Recombinant Vascular Endothelial Growth Factor Secreted From Tissue-Engineered Bioartificial Muscles Promotes Localized Angiogenesis , 2001, Circulation.

[41]  U. Lendahl,et al.  Generalized potential of adult neural stem cells. , 2000, Science.

[42]  A. Rosenthal,et al.  Embryonic Stem Cells Go Dopaminergic , 2000, Neuron.

[43]  J. Kimble,et al.  Functional domains of LAG-2, a putative signaling ligand for LIN-12 and GLP-1 receptors in Caenorhabditis elegans. , 1997, Molecular biology of the cell.

[44]  A. Hinek,et al.  Bioengineering of elastic cartilage with aggregated porcine and human auricular chondrocytes and hydrogels containing alginate, collagen, and kappa-elastin. , 1999, Journal of biomedical materials research.

[45]  N. Shah,et al.  Integration of multiple instructive cues by neural crest stem cells reveals cell-intrinsic biases in relative growth factor responsiveness. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[46]  M. Tuszynski,et al.  Neurotrophic factors, gene therapy, and neural stem cells for spinal cord repair , 2002, Brain Research Bulletin.

[47]  N. Perrimon,et al.  Drosophila unpaired encodes a secreted protein that activates the JAK signaling pathway. , 1998, Genes & development.

[48]  Gord Fishell,et al.  The role of notch in promoting glial and neural stem cell fates. , 2002, Annual review of neuroscience.

[49]  U. Lendahl,et al.  Oh no, Notch again! , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[50]  S. Artavanis-Tsakonas,et al.  Notch Signaling : Cell Fate Control and Signal Integration in Development , 1999 .

[51]  P. Rakic,et al.  Processing of the notch ligand delta by the metalloprotease Kuzbanian. , 1999, Science.

[52]  D. Ward,et al.  Human ligands of the Notch receptor. , 1999, The American journal of pathology.

[53]  Christel Brou,et al.  Signalling downstream of activated mammalian Notch , 1995, Nature.

[54]  A. Kiger,et al.  Stem Cell Self-Renewal Specified by JAK-STAT Activation in Response to a Support Cell Cue , 2001, Science.

[55]  D. Urry,et al.  Elastic Protein‐based Materials in Tissue Reconstruction a , 1997, Annals of the New York Academy of Sciences.

[56]  T. Jessell,et al.  Progression from Extrinsic to Intrinsic Signaling in Cell Fate Specification A View from the Nervous System , 1999, Cell.

[57]  T. Kadesch,et al.  Notch signaling: a dance of proteins changing partners. , 2000, Experimental cell research.

[58]  A. Björklund,et al.  Cell replacement therapies for central nervous system disorders , 2000, Nature Neuroscience.

[59]  J. Hubbell,et al.  Molecular properties of fibrin-based matrices for promotion of angiogenesis in vitro. , 2001, Microvascular research.

[60]  S. Cohen,et al.  Glycosyltransferase activity of Fringe modulates Notch–Delta interactions , 2000, Nature.

[61]  M. Rao,et al.  Properties of a fetal multipotent neural stem cell (NEP cell). , 2002, Developmental biology.

[62]  C. Morrow,et al.  Production and Purification of a Recombinant Elastomeric Polypeptide, G‐(VPGVG)19‐VPGV, from Escherichia coli , 1992, Biotechnology progress.

[63]  R. Sidman,et al.  Engraftable human neural stem cells respond to development cues, replace neurons, and express foreign genes , 1998, Nature Biotechnology.

[64]  I. Weissman,et al.  Translating stem and progenitor cell biology to the clinic: barriers and opportunities. , 2000, Science.

[65]  Y. Bae,et al.  Bioactive Polymers for Biohybrid Artificial Pancreas , 2001, Journal of drug targeting.

[66]  Howard Einspahr,et al.  Crystal structure and conformation of the cyclic trimer of a repeat pentapeptide of elastin, cyclo-(L-valyl-L-prolylglycyl-L-valylglycyl)3 , 1980 .

[67]  R. Bakay,et al.  Fetal tissue grafting into the central nervous system: yesterday, today, and tomorrow. , 1993, Neurosurgery.

[68]  I. Bernstein,et al.  Immobilization of Notch ligand, Delta-1, is required for induction of notch signaling. , 2000, Journal of cell science.

[69]  B. Dickson,et al.  Crossing the Midline Roles and Regulation of Robo Receptors , 2000, Neuron.

[70]  David J Anderson,et al.  Transient Notch Activation Initiates an Irreversible Switch from Neurogenesis to Gliogenesis by Neural Crest Stem Cells , 2000, Cell.

[71]  David Hanson.  Pesticide Makers Making Headway With Risk Assessment Argument , 1994 .

[72]  A. Bigas,et al.  Notch1 and Notch2 Inhibit Myeloid Differentiation in Response to Different Cytokines , 1998, Molecular and Cellular Biology.

[73]  F. Jiménez,et al.  The EGF and FGF Receptors Mediate Neuroglian Function to Control Growth Cone Decisions during Sensory Axon Guidance in Drosophila , 2000, Neuron.

[74]  B. Trask,et al.  The human homolog of rat Jagged1 expressed by marrow stroma inhibits differentiation of 32D cells through interaction with Notch1. , 1998, Immunity.

[75]  T. Jessell,et al.  A Homeodomain Protein Code Specifies Progenitor Cell Identity and Neuronal Fate in the Ventral Neural Tube , 2000, Cell.

[76]  U. Lendahl,et al.  The origin of the ankyrin repeat region in Notch intracellular domains is critical for regulation of HES promoter activity , 2001, Mechanisms of Development.

[77]  C. Macosko,et al.  Elastomeric polypentapeptides cross-linked into matrixes and fibers. , 2001, Biomacromolecules.

[78]  David J. Anderson,et al.  Glial growth factor restricts mammalian neural crest stem cells to a glial fate , 1994, Cell.

[79]  N. Baker,et al.  Notch signaling in the nervous system. Pieces still missing from the puzzle. , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[80]  T. Yamaoka,et al.  Design and Biosynthesis of Elastin-like Artificial Extracellular Matrix Proteins Containing Periodically Spaced Fibronectin CS5 Domains , 1999 .

[81]  M. Apuzzo,et al.  Utilization of unilateral and bilateral stereotactically placed adrenomedullary-striatal autografts in parkinsonian humans: rationale, techniques, and observations. , 1990, Neurosurgery.

[82]  David J. Anderson,et al.  Regulatory Mechanisms in Stem Cell Biology , 1997, Cell.

[83]  C. Goodman,et al.  Chimeric Axon Guidance Receptors The Cytoplasmic Domains of Slit and Netrin Receptors Specify Attraction versus Repulsion , 1999, Cell.

[84]  D. Anderson,et al.  Stem Cells and Pattern Formation in the Nervous System The Possible versus the Actual , 2001, Neuron.

[85]  Christopher W. Macosko,et al.  Mechanical Properties of Cross-Linked Synthetic Elastomeric Polypentapeptides , 2001 .

[86]  M. Tessier-Lavigne,et al.  Hierarchical Organization of Guidance Receptors: Silencing of Netrin Attraction by Slit Through a Robo/DCC Receptor Complex , 2001, Science.

[87]  David J. Anderson,et al.  The bHLH Transcription Factors OLIG2 and OLIG1 Couple Neuronal and Glial Subtype Specification , 2002, Cell.

[88]  Tasuku Honjo,et al.  Notch1 and Notch3 Instructively Restrict bFGF-Responsive Multipotent Neural Progenitor Cells to an Astroglial Fate , 2001, Neuron.

[89]  Jie Xu,et al.  Product purification by reversible phase transition following Escherichia coli expression of genes encoding up to 251 repeats of the elastomeric pentapeptide GVGVP. , 1996, Protein expression and purification.

[90]  R. Thurow.  Oh, NO! , 1984, The Angle orthodontist.

[91]  Tao Sun,et al.  Common Developmental Requirement for Olig Function Indicates a Motor Neuron/Oligodendrocyte Connection , 2002, Cell.