Neural Stem Cells and Neural Repair

Principal Investigator

Dr. Charo Sánchez Pernaute
rpernaute@inbiomed.org

Dr Sanchez-Pernaute is a neurologist and neuroscientist with extensive experience in cellular and molecular therapies for Parkinson Disease, as well as in embryonice stem cells and dopamine differentiation. After completing her MD and residency in Neurology, she obtained her PhD at University Autonoma de Madrid under the supervision of Dr Garcia de Yebenes, working on experimental models of Parkinson Disease. In 1998 she moved to the US National Institutes of Health for a post doc in the laboratory of Krys Bankiewicz, at the National Institute for Neurological Disorders and Stroke (NINDS) and started working on DA embryonic stem cells in collaboration with McKay's lab. In 2000 she moved to Harvard University where she continued her work at McLean Hospital in Ole Isacson's lab. She was promoted to Assistant Professor in 2004 and in 2005 joined the Harvard Stem Cell Institute. During this time she has continued working on primate and human embryonic stem cells to generate DA neurons for transplantation and functional restoration in models of Parkinson Disease. She also kept her long-standing interest and research work on functional neuroimaging (PET and fMRI) for in vivo evaluation of novel therapeutic approaches in experimental models of Parkinson Disease and the study of disease mechanisms. In December 2007 she joined Inbiomed to direct the cell therapy program for Parkinson Disease.

Scientific interest

Our aim is to generate dopamine neurons that have a correct midbrain identity and functional capacity from pluripotent stem cells. We hope that in the future such stem cell derived neurons could be applied to the treatment of patients with Parkinson Disease.

From previous work we know that fetal ventral midbrain neuroblasts transplanted into the brain of patients with Parkinson disease, survive for many years and are able to restore synaptic connections and improve motor function (Méndez et al 2005). However, this approach is limited by the scarcity of fetal tissue and the difficulty that poses for standardization. Pluripotent stem cells are broadly regarded as an alternative cell source for cell-based therapies, with the advantges of availability and amenability to extensive in vitro culture.

Why do we use pluripotent or embryonic stem cells? Pluripotent stem cells are an ideal cell source for generating quality DA neurons of the correct phenotype and in sufficient numbers for cell replacement and other biomedical applications, including high throughput pharmacological screens and disease models. Our interest is focused in synaptic restoration that requires the in stem cell derived neurons to have a correct midbrain phenotype and functional capacity in vivo. We can obtain this specific DA phenotype from pluripotent stem cells only -adult neural stem cells have limited differentiation capacity (lineage restriction). Furthermore, through direct cell reprogramming we are now able to obtain neurons from patients with familial and sporadic forms of Parkinson disease that will allow us to develop predictive in vitro models to evaluate disease modifying therapies.

In our current projects we are characterizing a number of factors that control cell fate and specification in the midbrain during hES differentiation. We are investigating strategies to enrich and select our target population based on expression of reporter genes under the control of specific transcription factors. This will allow us to obtain adequate cell populations for transplantation, minimizing the risk of proliferation and optimizing the efficiency. We use rodent models of Parkinson Disease to validate the functional potency of these neurons in vivo. In other project we are investigating the role of dopamine receptors and other factors in the development of dyskinesia, an invalidating complication of dopamine replacement therapies.

Staff

Postdoc Investigator:
Rakel López de Maturana

PhD Student(FPI, MICINN):
Julio César Aguila Benitez

Technicians:
Nerea Vázquez
Amaia Sousa

Summer students:
Aizea Morales
Ana Martínez Riaño

Publications

Cell Therapy:

Sánchez Pernaute R, Lee H, Yoshizaki T, Patterson M, Reske-Nielsen C, Sonntag KC, Studer L, Isacson O. Parthenogenetic dopamine neurons from primate ES cells restore function in experimental Parkinson disease. Brain. 2008 Aug;131(Pt 8):2127-39. Epub 2008 Jul 22.
Pruszak J, Sonntag KC, Aung MH, Sanchez-Pernaute R, Isacson O. Markers and methods for cell sorting of human embryonic stem cell-derived neural cell populations. Stem Cells 2007; 25:2257-68.
Sonntag KC, Pruszak J, Yoshizaki T, van Arensbergen J, Sanchez-Pernaute R, Isacson O. Enhanced yield of neuroepithelial precursors and midbrain-like dopaminergic neurons from human embryonic stem cells using the BMP antagonist Noggin. Stem Cells 2007; 25:411-8.
Sonntag KC, Sánchez Pernaute R. Tailoring human embryonic stem cells for therapy development in neurodegenerative diseases. Current Opin Investig Drugs 2006; 7:614-.8
Ferrari D, Sánchez-Pernaute R*, Lee H, Studer L, Isacson O. Transplanted dopamine neurons derived from primate ES cells preferentially innervate DARPP-32 striatal progenitors. EJN. 2006; 24:1885-96.
Bankiewicz KS, Forsayeth JR, Eberling JL, Sanchez-Pernaute R, Pivirotto P, Bringas J, Herscovitch P, Carson RE, Eckelman WC, Reutter B, Cunningham J. Long-term Clinical Improvement in MPTP-lesioned Primates after Gene Therapy with AAV-hAADC. Mol Ther. 2006; 14:564-70
Mendez I, Sánchez-Pernaute R, Cooper O, Viñuela A, Ferrari D, Bjorklund L, Dagher A, Isacson O. Cell type analysis of fetal dopamine cell suspension transplants in striatum and substantia nigra of patients with Parkinson’s disease. Brain 2005; 128:1498-1510
Sánchez-Pernaute R*, Studer L, Ferrari D, Perrier A, Lee H, Vinuela A, Isacson O. Long-term survival of dopamine neurons derived from parthenogenetic primate embryonic stem cells (Cyno1) in rat and primate striatum. Stem Cells 2005; 23:914-922.
Kim JH, Auerbach JM, Rodriguez-Gomez JA, Velasco I, Gavin D, Lumelsky N, Lee SH, Nguyen JB, Sánchez Pernaute R, Bankiewicz K, McKay R. Dopaminergic midbrain neurons derived from embryonic stem cells function in an animal model of Parkinson's disease. Nature 2002; 418:50-56.
Bjorklund LM, Sánchez-Pernaute R, Chung S, Andersson T, Chen IY, McNaught KS, Brownell AL, Jenkins BG, Wahlestedt C, Kim KS, Isacson O. Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model. Proc Natl Acad Sci U. S. A. 2002; 99:2344-2349.
Fontan A*, Rojo A*, Sánchez Pernaute R*, Hernandez I, Lopez I, Castilla C, Sánchez Albisua J, Perez Higueras A, Al-Rashid I, Rabano A, Gonzalo I, Angeles Mena M, Cools A, Eshuis S, Maguire P, Pruim J, Leenders K, Garcia de Yebenes J. Effects of fibroblast growth factor and glial-derived neurotrophic factor on akinesia, F-DOPA uptake and dopamine cells in parkinsonian primates. Parkinsonism Relat Disord 2002; 8:311-323
Isacson O, Bjorklund L, Sanchez-Pernaute R. Parkinson's disease: interpretations of transplantation study are erroneous. Nat Neurosci. 2001;4:553.
Sánchez-Pernaute R, Harvey-White J, Cunnigham J, Bankiewicz KS. Functional effect of adeno-associated virus mediated gene transfer of aromatic L-amino acid decarboxylase into the striatum of 6OHDA-lesioned rats. Mol Ther 2001;4:324-330.
Nguyen JB, Sánchez-Pernaute R, Cunnigham J, Bankiewicz KS. Convection-enhanced delivery of AAV-2 combined with heparin increases TK gene transfer in the rat brain. Neuroreport 2001;12:1961-1964.
Sánchez-Pernaute R, Studer L, Bankiewicz KS, Major EO, McKay RDG. In vitro generation and transplantation of precursor-derived human dopamine neurons. J Neurosci Res 2001; 65:284-288.

Functional models of Parkinson Disease:

Sánchez Pernaute R, Wang J-Q, Kuruppu D, Cao L, Tueckmantel W, Kozikowski A, Isacson O, Brownell A-L. Enhanced Binding of Metabotropic Glutamate Receptor Type 5 (mGluR5) PET Tracers in the Brain of Parkinsonian Primates. Neuroimage. 2008 Aug 1;42(1):248-51. Epub 2008 Apr 20.
Viaro R, Sanchez-Pernaute R, Marti M, Trapella C, Isacson O, Morari M.Nociceptin/orphanin FQ receptor blockade attenuates MPTP-induced parkinsonism.Neurobiol Dis. 2008 Jun;30(3):430-8. Epub 2008 Mar 8.
Sánchez Pernaute R*, Jenkins BG, Choi J-K, Chen IYC, Isacson O. In vivo evidence of dopamine receptor sensitization in parkinsonian rodents and primates with L-DOPA induced dyskinesias. Neurobiol Dis 2007;27:220-7.
Sánchez Pernaute R, Brownell AL, Jenkins BG, Isacson O. Insights into Parkinson’s disease models and neurotoxicity using non-invasive imaging. Toxicology and Applied Pharmacology. 2005; 207 (S2): 251-6.
Jenkins BG, Sánchez-Pernaute R, Brownell AL, Chen YI, Isacson O. Mapping dopamine function in primates using pharmacological MRI. J Neurosci 2004; 24:9553-60.
Sánchez-Pernaute R, Ferree A, Cooper O, Yu M, Brownell AL, and Isacson O. Selective COX-2 inhibition prevents progressive dopamine neuron degeneration in a rat model of Parkinson’s Disease. Journal of Neuroinflammation 2004; 1:6-13.
Oiwa Y, Sánchez-Pernaute R, Harvey-White J, Bankiewicz KS. Progressive and extensive dopaminergic degeneration induced by convection-enhanced delivery of 6-hydroxydopamine into the rat striatum: a novel rodent model of Parkinson disease. J Neurosurg 2003;98:136-44.
Brownell AL, Canales K, Chen YI, Jenkins BG, Owen C, Livni E, Yu M, Cicchetti F, Sanchez-Pernaute R, Isacson O. Mapping of brain function after MPTP-induced neurotoxicity in a primate Parkinson's disease model. Neuroimage 2003;20:1064-75.