Reversal of Dopaminergic Degeneration in a Parkinsonian Rat following Micrografting of Human Bone Marrow-Derived Neural Progenitors

Parkinson's disease (PD) is a common neurodegenerative disease characterized by the selective loss of dopaminergic (DA) neurons in the midbrain. Various types of stem cells that have potential to differentiate into DA neurons are being investigated as cellular therapies for PD. Stem cells also secrete growth factors and therefore also may have therapeutic effects in promoting the health of diseased DA neurons in the PD brain. To address this possibility in an experimental model of PD, bone marrow-derived neuroprogenitor-like cells were generated from bone marrow procured from healthy human adult volunteers and their potential to elicit recovery of damaged DA axons was studied in a partial lesion rat model of PD. Following collection of bone marrow, mesenchymal stem cells (MSC) were isolated and then genetically modified to create SB623 cells by transient transfection with the intracellular domain of the Notch1 gene (NICD), a modification that upregulates expression of certain neuroprogenitor markers. Ten deposits of 0.5 μl of SB623 cell suspension adjusted from 6,000 to 21,000 cells/μl in PBS or PBS alone were stereotaxically placed in the striatum 1 week after the nigrostriatal projection had been partially lesioned in adult F344 rats by injection of 6-hydroxydopamine (6-OHDA) into the striatum. At 3 weeks, a small number of grafted SB623 cells survived in the lesioned striatum as visualized by expression of the human specific nuclear matrix protein (hNuMA). In rats that received SB623 cells, but not in control rats, dense tyrosine hydroxylase immunoreactive (TH-ir) fibers were observed around the grafts. These fibers appeared to be rejuvenated host DA axons because no TH-ir in soma of surviving SB623 cells or coexpression of TH and hNuMA-ir were observed. In addition, dense serotonin immunoreactive (5-HT-ir) fibers were observed around grafted SB623 cells and these fibers also appeared to be of the host origin. Also, in some SB623 grafted rats that were sacrificed within 2 h of dl-amphetamine injection, hot spots of c-Fos-positive nuclei that coincided with rejuvenated dense TH fibers around the grafted SB623 cells were observed, suggesting increased availability of DA in these locations. Our observations suggest that NICD-transfected MSC hold potential as a readily available autologous or allogenic cellular therapy for ameliorating the degeneration of DA and 5-HT neurons in PD patients.

[1]  J. Kocsis,et al.  Remyelination of the Rat Spinal Cord by Transplantation of Identified Bone Marrow Stromal Cells , 2002, The Journal of Neuroscience.

[2]  M. Sofroniew,et al.  Trophism, Transplantation, and Animal Models of Parkinson's Disease , 1993, Experimental Neurology.

[3]  T. Dawson,et al.  Gene therapy to the rescue in Parkinson's disease. , 2001, Trends in pharmacological sciences.

[4]  B. Davidson,et al.  Dopaminergic Neurons Protected from Degeneration by GDNF Gene Therapy , 1997, Science.

[5]  T. Palmer,et al.  Vascular niche for adult hippocampal neurogenesis , 2000, The Journal of comparative neurology.

[6]  C. Shih,et al.  Derivation of neural stem cells from mesenchymal stemcells: evidence for a bipotential stem cell population. , 2008, Stem cells and development.

[7]  Catherine M. Verfaillie,et al.  Pluripotency of mesenchymal stem cells derived from adult marrow , 2002, Nature.

[8]  M. Chopp,et al.  Human bone marrow stromal cell cultures conditioned by traumatic brain tissue extracts: Growth factor production , 2002, Journal of neuroscience research.

[9]  M. Kameda,et al.  Comparison of the therapeutic potential of adult and embryonic neural precursor cells in a rat model of Parkinson disease. , 2008, Journal of neurosurgery.

[10]  G. Castelo-Branco,et al.  Persephin-Overexpressing Neural Stem Cells Regulate the Function of Nigral Dopaminergic Neurons and Prevent Their Degeneration in a Model of Parkinson's Disease , 2002, Molecular and Cellular Neuroscience.

[11]  B. Davidson,et al.  Differential effects of glial cell line-derived neurotrophic factor (GDNF) in the striatum and substantia nigra of the aged Parkinsonian rat , 1999, Gene Therapy.

[12]  R. Hauser,et al.  Fetal nigral grafts survive and mediate clinical benefit in a patient with Parkinson's disease , 1998, Movement disorders : official journal of the Movement Disorder Society.

[13]  Jacques Galipeau,et al.  Therapeutic angiogenesis using autologous bone marrow stromal cells: improved blood flow in a chronic limb ischemia model. , 2003, The Annals of thoracic surgery.

[14]  Sl Harris.  Amphetamine and cocaine induce drug-specific activation of the c-fos gene in striosome-matrix compartment and limbic subdivisions of the striatum , 1990 .

[15]  M. Chopp,et al.  Human marrow stromal cell therapy for stroke in rat: Neurotrophins and functional recovery , 2002, Neurology.

[16]  A. Graybiel,et al.  Amphetamine and cocaine induce drug-specific activation of the c-fos gene in striosome-matrix compartments and limbic subdivisions of the striatum. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Jenner,et al.  Effects of 6-hydroxydopamine lesions of the nigrostriatal pathway on striatal serotonin innervation in adult rats , 1991, Brain Research.

[18]  M. Beal,et al.  Functional engraftment of human ES cell–derived dopaminergic neurons enriched by coculture with telomerase-immortalized midbrain astrocytes , 2006, Nature Medicine.

[19]  Y. Ishizuka,et al.  Serotonergic activity in the rat striatum after intrastriatal transplantation of fetal nigra as measured by microdialysis , 1998, Brain Research.

[20]  S. Kurpad,et al.  In vitro and in vivo characterization of neurally modified mesenchymal stem cells induced by epigenetic modifiers and neural stem cell environment. , 2008, Stem cells and development.

[21]  D J Prockop,et al.  Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Mari Dezawa,et al.  Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation. , 2004, The Journal of clinical investigation.

[23]  D. Guilloteau,et al.  Partial recovery of dopaminergic pathway after graft of adult mesenchymal stem cells in a rat model of Parkinson's disease , 2008, Neurochemistry International.

[24]  J. Sanchez-Ramos.  Neural cells derived from adult bone marrow and umbilical cord blood , 2002, Journal of neuroscience research.

[25]  A. Caplan,et al.  Mesenchymal stem cells as trophic mediators , 2006, Journal of cellular biochemistry.

[26]  W. H. Oertel,et al.  Progressive degeneration of nigrostriatal dopamine neurons following intrastriatal terminal lesions with 6-hydroxydopamine: A combined retrograde tracing and immunocytochemical study in the rat , 1994, Neuroscience.

[27]  J. Kordower,et al.  Gene therapy to the rescue in Parkinson's disease. Response from Kordower and Aebischer. , 2001, Trends in pharmacological sciences.

[28]  C. Verfaillie,et al.  Human Bone Marrow Stem Cells Exhibit Neural Phenotypes and Ameliorate Neurological Deficits after Grafting into the Ischemic Brain of Rats , 2002, Experimental Neurology.

[29]  R. Sandyk,et al.  Serotonin in involuntary movement disorders. , 1988, The International journal of neuroscience.

[30]  M. Chopp,et al.  Intracerebral transplantation of marrow stromal cells cultured with neurotrophic factors promotes functional recovery in adult rats subjected to traumatic brain injury. , 2002, Journal of neurotrauma.

[31]  P. Jenner,et al.  3,4-Methylenedioxymethamphetamine (Ecstasy) Inhibits Dyskinesia Expression and Normalizes Motor Activity in 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine-Treated Primates , 2003, The Journal of Neuroscience.

[32]  C. Marsden,et al.  Serotonergic sprouting in the neostriatum after intrastriatal transplantation of fetal ventral mesencephalon , 1991, Brain Research.

[33]  D. Hess,et al.  Transplantation of Bone Marrow-Derived Stem Cells: A Promising Therapy for Stroke , 2007, Cell transplantation.

[34]  J. Bloch,et al.  Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease. , 2000, Science.

[35]  Yi Li,et al.  Treatment of neural injury with marrow stromal cells , 2002, The Lancet Neurology.

[36]  Yi Li,et al.  Intravenous Administration of Human Bone Marrow Stromal Cells Induces Angiogenesis in the Ischemic Boundary Zone After Stroke in Rats , 2003, Circulation research.

[37]  M. Dezawa.  Insights into autotransplantation: the unexpected discovery of specific induction systems in bone marrow stromal cells , 2006, Cellular and Molecular Life Sciences CMLS.

[38]  D. Gash,et al.  Adrenal medulla grafts enhance recovery of striatal dopaminergic fibers. , 1987, Science.

[39]  C. Verfaillie.  Adult stem cells: assessing the case for pluripotency. , 2002, Trends in cell biology.

[40]  K. Lachapelle,et al.  Postnatal bone marrow stromal cells elicit a potent VEGF-dependent neoangiogenic response in vivo , 2003, Gene Therapy.

[41]  N. Bresolin,et al.  Induction of Neurotrophin Expression via Human Adult Mesenchymal Stem Cells: Implication for Cell Therapy in Neurodegenerative Diseases , 2007, Cell transplantation.

[42]  Esther Lipokatic-Takacs,et al.  Fibroblast Growth Factor (FGF)-2 and FGF Receptor 3 Are Required for the Development of the Substantia Nigra, and FGF-2 Plays a Crucial Role for the Rescue of Dopaminergic Neurons after 6-Hydroxydopamine Lesion , 2007, The Journal of Neuroscience.

[43]  A. Manira,et al.  Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[44]  G. M. Halliday,et al.  Loss of brainstem serotonin- and substance P-containing neurons in Parkinson's disease , 1990, Brain Research.

[45]  M. Chopp,et al.  Intracerebral transplantation of bone marrow stromal cells in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease , 2001, Neuroscience Letters.

[46]  S. Dunnett,et al.  Graft-derived recovery from 6-OHDA lesions: specificity of ventral mesencephalic graft tissues , 2004, Experimental Brain Research.

[47]  S. Carbonetto,et al.  Nerve fiber growth on defined hydrogel substrates. , 1982, Science.

[48]  M. Oka,et al.  Peripheral nerve regeneration along collagen filaments , 2001, Brain Research.

[49]  E. Oldfield,et al.  Reversal of hemiparkinsonian syndrome in nonhuman primates by amnion implantation into caudate nucleus. , 1994, Journal of neurosurgery.

[50]  J. Kordower,et al.  Human neural progenitors deliver glial cell line-derived neurotrophic factor to parkinsonian rodents and aged primates , 2006, Gene Therapy.

[51]  C. Svendsen,et al.  Human neural progenitor cells over-expressing IGF-1 protect dopamine neurons and restore function in a rat model of Parkinson's disease , 2008, Experimental Neurology.

[52]  R. McKay,et al.  Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease , 2002, Nature.

[53]  T. Itakura,et al.  Protection of dopamine neurons by bone marrow stromal cells , 2007, Brain Research.

[54]  F. Gage,et al.  Nurr1, an orphan nuclear receptor, is a transcriptional activator of endogenous tyrosine hydroxylase in neural progenitor cells derived from the adult brain. , 1999, Development.

[55]  L. Iacovitti,et al.  Studies on the Differentiation of Dopaminergic Traits in Human Neural Progenitor Cells in Vitro and in Vivo , 2004, Cell transplantation.

[56]  J. Kapfhammer,et al.  Basic Fibroblast Growth Factor Modulates Density of Blood Vessels and Preserves Tight Junctions in Organotypic Cortical Cultures of Mice: A New In Vitro Model of the Blood–Brain Barrier , 2007, The Journal of Neuroscience.

[57]  I. Black,et al.  Adult rat and human bone marrow stromal cells differentiate into neurons , 2000, Journal of neuroscience research.

[58]  C D Marsden,et al.  Grafts of fetal dopamine neurons survive and improve motor function in Parkinson's disease. , 1990, Science.

[59]  D. Snow,et al.  Interactions of developing neurons with the extracellular matrix , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[60]  Michael Chopp,et al.  Marrow Stromal Cell Transplantation after Traumatic Brain Injury Promotes Cellular Proliferation within the Brain , 2004, Neurosurgery.

[61]  T. Schallert,et al.  Delivery of a GDNF Gene into the Substantia Nigra after a Progressive 6-OHDA Lesion Maintains Functional Nigrostriatal Connections , 2000, Experimental Neurology.

[62]  K. Houkin,et al.  Intravenous administration of glial cell line‐derived neurotrophic factor gene‐modified human mesenchymal stem cells protects against injury in a cerebral ischemia model in the adult rat , 2006, Journal of neuroscience research.

[63]  R. Vandlen,et al.  Mesencephalic dopaminergic neurons protected by GDNF from axotomy-induced degeneration in the adult brain , 1995, Nature.

[64]  Masahiko Takano,et al.  Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone‐marrow stromal cells , 2001, The European journal of neuroscience.

[65]  M. Chopp,et al.  Coupling of Angiogenesis and Neurogenesis in Cultured Endothelial Cells and Neural Progenitor Cells after Stroke , 2008, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[66]  M. Chopp,et al.  Combination therapy of stroke in rats with a nitric oxide donor and human bone marrow stromal cells enhances angiogenesis and neurogenesis , 2004, Brain Research.

[67]  T. Yasuhara,et al.  Transplantation of Human Neural Stem Cells Exerts Neuroprotection in a Rat Model of Parkinson's Disease , 2006, The Journal of Neuroscience.

[68]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[69]  R. Bartus,et al.  Striatal delivery of neurturin by CERE-120, an AAV2 vector for the treatment of dopaminergic neuron degeneration in Parkinson's disease. , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[70]  Ora Dillon-Carter,et al.  Bone marrow grafts restore cerebral blood flow and blood brain barrier in stroke rats , 2004, Brain Research.

[71]  R. Lindsay,et al.  Brain‐Derived Neurotrophic Factor Protects Dopamine Neurons Against 6‐Hydroxydopamine and N‐Methyl‐4‐Phenylpyridinium Ion Toxicity: Involvement of the Glutathione System , 1992, Journal of neurochemistry.

[72]  K. Then,et al.  Generating neuron-like cells from BM-derived mesenchymal stromal cells in vitro. , 2007, Cytotherapy.

[73]  A. Asawachaicharn,et al.  Human mesenchymal stem cell subpopulations express a variety of neuro-regulatory molecules and promote neuronal cell survival and neuritogenesis , 2006, Experimental Neurology.

[74]  Gwang Lee,et al.  Mesenchymal stem cells therapy exerts neuroprotection in a progressive animal model of Parkinson’s disease , 2008, Journal of neurochemistry.