The Morris K. Udall Center of Excellence for Parkson's Disease Research at Columbia University

Director: Robert E. Burke, MD

Center Title: Mechanisms of dopamine neuron degeneration

Central Theme

To understand the molecular and cellular mechanisms of neuron degeneration in Parkinson’s disease.

Central Structure

The Udall Center at Columbia University was competitively renewed for 2009-2014, and has now completed Year 01 of the renewal award. Since this is the first Progress Report for the renewal award, we will provide in this section a detailed description of the new structure.

The greatest challenge posed by Parkinson’s disease (PD) is to develop therapies that address the underlying degenerative process. The overriding theme of the Udall Center at Columbia is to address this challenge. Our efforts have two guiding principles. The first is that development of such therapies ultimately depends on a better understanding of mechanisms of disease. The second is that patients cannot wait for a full understanding of this disease to be in hand before efforts are made to translate new knowledge into treatments. Based on these principles, our Center now consists of four Projects and five Cores that will be integrated according to five current central themes in the pathogenesis of PD. Project 1 (Drs. David Sulzer, PhD, and Ana Maria Cuervo, MD, PhD): "Roles for cytosolic dopamine in PD pathogenesis" will extend a line of investigation that suggests that "multiple hits", consisting of mishandling of 1) ?-synuclein (?-syn) degradation, 2) cytosolic calcium, and 3) cytosolic dopamine (DA), underlie PD. Project 2 (Dr. William Dauer, MD): "The FADD/Caspase-8 pathway as a mediator of LRRK2 neurotoxicity" proposes to test whether signaling through FADD/caspase-8 is required for the nigrostriatal-related phenotypes seen in PD mutant LRRK2 transgenic mice, including reduced locomotion (L-dopa responsive), decreased striatal dopamine (DA) efflux, and axonal degeneration. Project 3 (Dr. Lloyd Greene, PhD): "Gene regulation in PD" is based on the rationale that neuron degeneration and death, irrespective of the initiating causes, require transcriptional induction of death-associated genes and that such genes and the pathways that are up- and down-stream of them are potential targets for therapeutic intervention in PD. Project 4 (Dr. Robert E. Burke, MD): "Translational approaches to neuroprotection of axons in PD". This Project is the translational Project of our Udall Center. We have developed a promising gene therapy approach that uses Akt, a kinase that protects neurons and their axons from degeneration. The goal of this Project is to refine this approach and to develop promising next-generation alternatives. Core A (Dr. Burke and Ms. Janice Savage) "Administrative Core" sets policies and standard operating procedures for Center activities, organizes and conducts regular internal and external scientific review, and organizes and implements all the necessary forms of scientific communication to make new discoveries and achievements within the Center known to the scientific community and the public. Core B (Dr. Burke) "Neuroscience Core" provides general laboratory services including maintenance and repair of all Core laboratory equipment used by the Principal Investigators. Core C: Brain Bank Core (Drs. Jean Paul Vonsattel and Lorraine Clark). In this renewal application, with its greater translational emphasis, the Udall Projects anticipated a greater need for brain tissue, and for tissue that has been characterized more extensively, both with regard to synuclein immunostaining and genetic analysis. This Core meets these new needs. In addition, this Core provides fully characterized brain tissue samples to the wider Udall and PD research communities. Core D: Core for Training in Translational Research (Dr. Burke). There is a desperate need to train young investigators in Neuroscience in the translational potential of their research. This need will be addressed by this training Core. This Core provides young MD or MD/PhD investigators who have trained in Neurology and then for one year in our Clinical Movement Disorders Fellowship Program full salary support for two years to train in the laboratory of one of the Udall Principal Investigators. Core E: PD Patient Fibroblast Core (Drs. Przedborski and Lorraine Clark) collects and stores skin fibroblasts from patients with sporadic PD and from healthy age-matched controls, destined for future genetic, epigenetic, and stem cell reprogramming investigations.

Recent Significant Advances

Mosharov EV, Larsen KE, Kanter E, Phillips KA, Wilson K, Schmitz Y, Krantz DE, Kobayashi K, Edwards RH, Sulzer D. Interplay between cytosolic dopamine, calcium, and alpha-synuclein causes selective death of substantia nigra neurons. Neuron 2009;62:218-229.

The basis for selective death of specific neuronal populations in PD remains unclear. PD is characterized by a preferential loss of dopaminergic neurons in the substantia nigra (SN), whereas neurons of the ventral tegmental area (VTA) are spared. Dr. Sulzer and his colleagues measured cytosolic dopamine (DA(cyt)) in cultured midbrain neurons, and confirmed that elevated DA(cyt) and its metabolites are neurotoxic and that genetic and pharmacological interventions that decrease DA(cyt) provide neuroprotection. L-DOPA increased DA(cyt) in SN neurons to levels 2- to 3-fold higher than in VTA neurons, a response dependent on dihydropyridine-sensitive Ca2+ channels, resulting in greater susceptibility of SN neurons to L-DOPA-induced neurotoxicity. DA(cyt) was not altered by alpha-synuclein deletion, although dopaminergic neurons lacking alpha-synuclein were resistant to L-DOPA-induced cell death. Thus, an interaction between Ca2+, DA(cyt), and alpha-synuclein may underlie the susceptibility of SN neurons in PD, suggesting multiple therapeutic targets.

Malagelada C, Jin ZH, Jackson-Lewis V, Przedborski S, Greene LA. Rapamycin protects against neuron death in in vitro and in vivo models of Parkinson's disease. J Neurosci 2010;30:1166-1175.

In an effort to link RTP801 induction to the mTOR pathway and to find a potential therapeutic approach to protecting neurons in PD, we found that rapamycin, a partial inhibitor of mTOR function blocks RTP801 induction by PD mimetics both in vitro and in vivo. Moreover, rapamycin protected neurons from PD mimetics both in vitro as well as in two different animal MPTP models. Our study also revealed that the mechanism by which rapamycin blocks RTP801 induction is by suppressing translation of its mRNA. Finally, our study examined the role of Akt signaling in the protective actions of rapamycin. We had previously reported that RTP801 mediates the inactivation of Akt in PD models and that this in turn was responsible for neuron death. Our findings showed that with rapamycin, the activity of Akt was preserved, again both in vivo and in vitro.

Available Resources

The mutant hR1441G LRRK2 BAC transgenic mice reported in 2009 have been transferred to and are available from the Jackson Laboratory Induced Mutant Resource Repository: http://www.jax.org/imr/index.html

During Year 01 of this award, in Core C: Brain Bank Core (Drs. Jean Paul Vonsattel and Lorraine Clark), 48 brains with Lewy pathology have been collected and banked. Genotyping by Dr. Clark is underway. Tissue material is available through the NYBB website: http://www.nybb.hs.columbia.edu/.

Plans for the Coming Year

Our successfully renewed Udall Center (2009-2014) will continue its work according to five current central themes in the pathogenesis of PD. Each of the Projects builds on discoveries in the genetics of PD. Project 1 (Sulzer & Cuervo) will continue their important work on the degradation of ?-synuclein. Projects 2 (Dauer) and 3 (Greene) will explore, in collaboration, mechanisms of LRRK2 toxicity. Project 4 (Burke) will utilize a new hLRRK2(R1441G) BAC transgenic model of PD, in which he has identified a dopaminergic axonopathy, in translational investigations. Project 1 will also examine relationships between processing of synuclein and two important components of the intracellular milieu of dopamine neurons: cytosolic dopamine and calcium. One of the central themes of our Udall Center has been to examine the role of programmed cell death in pathogenesis. This theme has taken on a new importance due to the work of Dr Dauer in Project 3, demonstrating an interaction of LRRK2 with proteins in the extrinsic cell death pathway. Independent lines of investigation in Projects 3 (Greene) and 4 (Burke) have converged on the importance of the survival signaling kinase Akt in the viability of dopamine neurons. The importance of chaperone-mediated autophagy in degradation of synuclein has been discovered in Project 1, and the role of macroautophagy in the maintenance of axons has emerged in Project 4. To support the emphasis in the current proposal on the translational implications of our work, the Center now includes a Brain Bank Core (Vonsattel & Clark), a Core for Training in Translational Neuroscience (Burke), and a Fibroblast Core (Przedborski) for the production of human pluripotent cells. Thus the Udall Center at Columbia is poised to discover new approaches to neuroprotection and restoration, and to move these discoveries to the clinic expeditiously.

Select Recent Publications

[Columbia Udall Principal Investigators are in BOLD]

Gubernator NG, Zhang H, Staal RG, Mosharov EV, Pereira DB, Yue M, Balsanek V, Vadola PA, Mukherjee B, Edwards RH, Sulzer D, Sames D. Fluorescent false neurotransmitters visualize dopamine release from individual presynaptic terminals. Science 2009;324:1441-1444.

Ho CC, Rideout HJ, Ribe E, Troy CM, Dauer WT. The Parkinson disease protein leucine-rich repeat kinase 2 transduces death signals via Fas-associated protein with death domain and caspase-8 in a cellular model of neurodegeneration. J Neurosci 2009;29:1011-1016.

Jorgensen ND, Peng Y, Ho CC, Rideout HJ, Petrey D, Liu P, Dauer WT. The WD40 domain is required for LRRK2 neurotoxicity. PLoS ONE 2009;4:e8463

Lu XH, Fleming SM, Meurers B, Ackerson LC, Mortazavi F, Lo V, Hernandez D, Sulzer D, Jackson GR, Maidment NT, Chesselet MF, Yang XW. Bacterial artificial chromosome transgenic mice expressing a truncated mutant parkin exhibit age-dependent hypokinetic motor deficits, dopaminergic neuron degeneration, and accumulation of proteinase K-resistant alpha-synuclein. J Neurosci 2009;29:1962-1976.

Mosharov EV, Larsen KE, Kanter E, Phillips KA, Wilson K, Schmitz Y, Krantz DE, Kobayashi K, Edwards RH, Sulzer D. Interplay between cytosolic dopamine, calcium, and alpha-synuclein causes selective death of substantia nigra neurons. Neuron 2009;62:218-229.

Oo TF, Marchionini DM, Yarygina O, O'Leary PD, Hughes RA, Kholodilov N, Burke RE. Brain-derived neurotrophic factor regulates early postnatal developmental cell death of dopamine neurons of the substantia nigra in vivo. Mol Cell Neurosci 2009;41:440-447.

Ries V, Cheng HC, Baohan A, Kareva T, Oo TF, Rzhetskaya M, Bland RJ, During MJ, Kholodilov N, Burke RE. Regulation of the postnatal development of dopamine neurons of the substantia nigra in vivo by Akt/protein kinase B. J Neurochem 2009;110:23-33.

Schmitz Y, Luccarelli J, Kim M, Wang M, Sulzer D. Glutamate controls growth rate and branching of dopaminergic axons. J Neurosci 2009;29:11973-11981.

Sproul AA, Xu Z, Wilhelm M, Gire S, Greene LA. Cbl negatively regulates JNK activation and cell death. Cell Res 2009;19:950-961.

Zhang H, Gubernator NG, Yue M, Staal RG, Mosharov EV, Pereira D, Balsanek V, Vadola PA, Mukherjee B, Edwards RH, Sulzer D, Sames D. Dopamine release at individual presynaptic terminals visualized with FFNs. J Vis Exp 2009;

Zhang W, Phillips K, Wielgus AR, Liu J, Albertini A, Zucca FA, Faust R, Qian SY, Miller DS, Chignell CF, Wilson B, Jackson-Lewis V, Przedborski S, Joset D, Loike J, Hong JS, Sulzer D, Zecca L. Neuromelanin Activates Microglia and Induces Degeneration of Dopaminergic Neurons: Implications for Progression of Parkinson's Disease. Neurotox Res 2009;

Bendor J, Lizardi-Ortiz JE, Westphalen RI, Brandstetter M, Hemmings HC, Jr., Sulzer D, Flajolet M, Greengard P. AGAP1/AP-3-dependent endocytic recycling of M5 muscarinic receptors promotes dopamine release. EMBO J 2010;29:2813-2826.

Biswas SC, Zhang Y, Iyirhiaro G, Willett RT, Rodriguez GY, Cregan SP, Slack RS, Park DS, Greene LA. Sertad1 plays an essential role in developmental and pathological neuron death. J Neurosci 2010;30:3973-3982.

Cheng HC, Burke RE. The Wld(S) mutation delays anterograde, but not retrograde, axonal degeneration of the dopaminergic nigro-striatal pathway in vivo. J Neurochem 2010;113:683-691.

Hnasko TS, Chuhma N, Zhang H, Goh GY, Sulzer D, Palmiter RD, Rayport S, Edwards RH. Vesicular glutamate transport promotes dopamine storage and glutamate corelease in vivo. Neuron 2010;65:643-656.

Lee M, Gubernator NG, Sulzer D, Sames D. Development of pH-responsive fluorescent false neurotransmitters. J Am Chem Soc 2010;132:8828-8830.

Malagelada C, Jin ZH, Jackson-Lewis V, Przedborski S, Greene LA. Rapamycin protects against neuron death in in vitro and in vivo models of Parkinson's disease. J Neurosci 2010;30:1166-1175.

Martinez-Vicente M, Talloczy Z, Wong E, Tang G, Koga H, Kaushik S, de Vries R, Arias E, Harris S, Sulzer D, Cuervo AM. Cargo recognition failure is responsible for inefficient autophagy in Huntington's disease. Nat Neurosci 2010;13:567-576.

Public Health Statement

Current therapies for PD treat only its symptoms, not its progression. The goal of our research is to use new knowledge about the mechanisms of neurodegeneration to develop therapies that will block disease progression. We use genetic and neurotoxin models to better understand these mechanisms. We realize that patients cannot wait for us to understand mechanisms completely before we try to develop new therapies. Therefore, we will also go forward with translational research to establish novel pharmacologic and gene therapies.

Other Developments of Interest

The first recipient of support through Training Core D as the Columbia Udall Center Postdoctoral Research Scientist, Dr. Oren Levy, has successfully applied for a K08 award. In year 02 support will be provided for the second recipient, Dr. Sheng-Han Kuo, to work in Dr. Sulzer’s Lab.

Dr. Sulzer and Dr. James Surmeier, Director of the Udall Center at Northwestern University, successfully applied for an Administrative Supplement to enable them to work together on a collaborative project. The overall goal of the work is to investigate the importance of macroautophagy in PD pathogenesis, a goal that is in keeping with one of the principal research themes of our Center. Dr. Sulzer’s specific role will be to determine the effect of mitochondrial oxidant stress on the turnover of mitochondria by autophagy. Dr. Sulzer will be assisted in this project by Dr. Sheng-Han Kuo, now the Columbia Udall Center Postdoctoral Research Scientist.