This proposal aims to assess in a relevant animal model the safety, feasibility and efficacy of adult stem cell administration to the CNS for treating neurological sequelae associated with lysosomal storage diseases. Specifically, adult stem cells derived from bone marrow, referred to as mesenchymal stem cells (MSCs) will be injected unilaterally into the caudate putamen of infant rhesus macaques (Macaca mulatta) using stereotaxic coordinates determined by MRI. Prior to and at various periods up to 1 year post-transplantation, the health, development, behavior as well as cognitive and motor skills of each transplant recipient will be evaluated using a battery of age appropriate tests and compared to sham-operated animals or normative control data. Additionally, the levels and anatomical distribution of male MSCs engrafted in the CNS of female transplant recipients will be evaluated using a real-time PCR assay we developed that targets sequences in the Macaca sp. Y chromosome. To confirm the real-time PCR data engrafted MSCs will be visualized in brain sections by immunofluorescent staining or FISH and counted using modern stereological techniques. Correlating MSC engraftment in brain with long-term affects on health, development, behavior and motor function will directly assess the safety and feasibility of intracranial MSC administration. Subsequently, MSCs will be injected into the CNS of infant macaques afflicted with globoid cell leukodystrophy (Krabbe's Disease). The latter is characterized by a lack of galactocerebrosidase (GALC) activity, which results in abnormal accumulation of storage material in oligodendrocytes causing progressive demyelination in the nervous system. In these studies MSC engraftment levels and distribution will be directly correlated with levels of expressed GALC mRNA and enzyme activity throughout the CNS. Furthermore, affects on disease progression will be evaluated using biochemical and histological methods as well as behavioral and motor performance tests. Consequently, this phase of the study will directly evaluate the therapeutic efficacy of MSC administration. Collectively, these proposed studies will yield important pre-clinical data in a relevant animal model that will be essential for developing stem cell-based therapies to treat neurological sequelae in human infants and children afflicted with various lysosomal storage diseases. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS052301-01A2
Application #
7257658
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Tagle, Danilo A
Project Start
2007-04-01
Project End
2011-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
1
Fiscal Year
2007
Total Cost
$560,910
Indirect Cost
Name
Tulane University
Department
Genetics
Type
Schools of Medicine
DUNS #
053785812
City
New Orleans
State
LA
Country
United States
Zip Code
70118
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Phinney, Donald G; Isakova, Iryna A (2014) Mesenchymal stem cells as cellular vectors for pediatric neurological disorders. Brain Res 1573:92-107
Isakova, Iryna A; Lanclos, Calvin; Bruhn, Julie et al. (2014) Allo-reactivity of mesenchymal stem cells in rhesus macaques is dose and haplotype dependent and limits durable cell engraftment in vivo. PLoS One 9:e87238
Haga, Christopher L; Phinney, Donald G (2012) MicroRNAs in the imprinted DLK1-DIO3 region repress the epithelial-to-mesenchymal transition by targeting the TWIST1 protein signaling network. J Biol Chem 287:42695-707
Boregowda, Siddaraju V; Krishnappa, Veena; Chambers, Jeremy W et al. (2012) Atmospheric oxygen inhibits growth and differentiation of marrow-derived mouse mesenchymal stem cells via a p53-dependent mechanism: implications for long-term culture expansion. Stem Cells 30:975-87
Lai, Wen-Tzu; Krishnappa, Veena; Phinney, Donald G (2011) Fibroblast growth factor 2 (Fgf2) inhibits differentiation of mesenchymal stem cells by inducing Twist2 and Spry4, blocking extracellular regulated kinase activation, and altering Fgf receptor expression levels. Stem Cells 29:1102-11
Isakova, Iryna A; Dufour, Jason; Lanclos, Calvin et al. (2010) Cell-dose-dependent increases in circulating levels of immune effector cells in rhesus macaques following intracranial injection of allogeneic MSCs. Exp Hematol 38:957-967.e1