Catecholamine Research in the 21st Century: Abstracts and Graphical Abstracts, 10th International Catecholamine Symposium, 2012

Preface

Lee E. Eiden

Section on Molecular Neuroscience, Laboratory of Cellular and Molecular Regulation, National Institutes of Health, Bethesda, MD, USA

The Tenth International Catecholamine Symposium (XICS) was held at The Asilomar Conference Grounds in Pacific Grove CA, September 9-13, 2012. It was the first international symposium focused on catecholamines in the 21st century, providing the title for the proceedings of the symposium found in this volume. David Goldstein, founding Chief of the Clinical Neurocardiology Section, NINDS and President of the 8th International Catecholamine Symposium, also held at Asilomar in 1996, provided the guiding inspiration for the XICS. Daniel O’Connor, Professor of Medicine and Pharmacology at the Institute for Genomic Medicine, University of California San Diego, and President of the Catecholamine Society was also President of the 10th International Catecholamine Symposium. David Sibley, Chief of the Molecular Neuropharmacology Section, NINDS, Bethesda, MD, Esther Sabban, Professor of Biochemisty and Molecular Biology, New York Medical College, and the editor of this volume constituted the XICS Executive Organizing Committee. Many others, most especially the participants who came from Austria, Canada, The Czech Republic, Denmark, France, Germany, Israel, Italy, Japan, Mexico, Norway, Russian, Taiwan, the United States and elsewhere, contributed importantly to the success of the XICS. The editor wishes to thank Dave Goldstein and Dan O'Connor especially for their devotion to the eventual completion of the Symposium and these proceedings.

Some of the broader topics most relevant to the current status of catecholamine research as a translational field have been covered in greater depth, by thought leaders in catecholamine research who were in attendance at the XICS, in a companion volume of Advances in Pharmacology. These include mechanisms of catecholamine biochemistry, cell biology, systems biology, clinical diagnosis, drug discovery and target discovery, and gene therapeutic treatment for catecholamine-related human disease. Contained herein are the comprehensive conference proceedings in the form of extended graphical abstracts, and some condensed abstracts, of almost all of the presentations at the meeting organized into the ten themes under which the symposium was convened. The themes are introduced as chapters each with a short introduction that attempts to identify the highlights and 'growth areas' within each. It is hoped that the reader will find these proceedings useful as a handbook for the current state of play of catecholamine research, until such time as the Proceedings of the 11th International Catecholamine Symposium succeeds them.

Theme A

Catecholamine Biosynthesis and Storage

Outline

Theme A Catecholamine Biosynthesis and Storage

Genetic Manipulation of Catecholamine Signaling in the Mouse

AADC Deficiency

Tyrosine Hydrolylase and Dopamine Beta-Hydroxylase

Pharmacokinetic and Pharmacodynamic Properties of Etamicastat, a New DBH Inhibitor

Estradiol-Mediated Regulation of Gene Expression of Catecholamine Biosynthetic Enzymes

Structural Basis for Regulation of Tyrosine Hydroxlyase by the Catecholamines

Unique Regulation of TH Gene Expression in Midbrain Dopamine Neurons

Intracellular Stability of Tyrosine Hydroxylase

The Peripheral Interaction of Tyrosine Hydroxylase and 14-3-3γ with Negatively Charged Phospholipidic Membranes

Dynamic Regulation of Tyrosine Hydroxylase Gene Expression by Key Fate-Determining Transcription Factors during Dopaminergic Neuronal Development in Vivo and in Vitro

Analysis of Tyrosine Hydroxylase Isoforms and Phosphorylation in Parkinson’s Disease

Non-dopaminergic Neurons Partly Expressing Dopaminergic Phenotype

Developing Brain as an Endocrine Organ

Brainstem DOPAergic System

Imaging Norepinephrine Transporters in Humans; Translational Research with PET

Imaging the Vesicular Monoamine Transporter (VMAT2) in Neurodegenerative Diseases

GTP Cyclohydrolase Regulation

Neonatal Diagnosis of Menkes Disease by a Pattern of Plasma Catechols

Catecholamine Metabolites Affected by the Copper-Dependent Enzyme Dopamine-Beta-Hydroxylase Provide Sensitive Blood and Csf Biomarkers for Viral Gene Therapy in Menkes Disease

Are the Enzymes of the Catecholamine Biosynthetic Pathway Locally Synthesized in the Axon?

Effects of Missense Mutations in Tyrosine Hydroxylase (TH) Found in Patients with Neurological Disorders Attributed to TH Deficiency

Tetrahydrobiopterin Deficiency Impairs Postnatal Increase of TH Protein via Insufficient Dopamine Biosynthesis

Selective Ablation of Dopamine Beta-Hydroxylase Neurons (Subpopulation of TH Neurons) in the Brain

Neural Circuit Mechanism for Learning Dependent on Dopamine Transmission

Regulation of Tyrosine Hydroxylase by Ser19-Phosphorylation-Dependent Binding to 14-3-3γ

Modeling the Dynamics of Dopamine Biosynthesis and its Regulation by Tyrosine Hydroxylase

Reassessment of Intrinsic Dopaminergic Innervation in the Human Enteric Nervous System – Clinical Implications

Midbrain Dopamine Neurons are Divided into Different Functional Groups

Cell-, Region- and Species-Specific Expression of VMAT2

Theme A Catecholamine Biosynthesis and Storage

Lee Eiden and David Goldstein

The major features of catecholamine biosynthesis [Tyrosine (TH) –> L-Dopa (AADC) –> DA (DBH) –> NE (PNMT) –> Epi] and storage [cytoplasmic DA, NE, Epi –> VMAT1,2 –> vesicular DA, NE, Epi] were worked out well before the end of the previous century. This chapter contains illustrative examples of progress since, and this is manifest in a more complete molecular understanding of the cell biology that allows TH to control CA biosynthesis; concrete clinical steps for detection and gene therapy for catecholamine deficiency diseases; and complete identification of the combinations of catecholamine biosynthetic enzymes and vesicular storage capacity that can be found throughout the brain and periphery, and distinctly in rodent and primate neuronal systems, from which a ‘post-classical’ view of catecholamine chemical neuroanatomy has emerged.

While TH retains pride of place as the rate limiting enzyme for catecholamine biosynthesis, new insights into the conversion of tyrosine to L-Dopa have been gained by fuller exploration of the role of GTP cyclohydrolase (GTPCH) in supplying the necessary co-factor, tetrahydrobiopterin, for this enzymatic conversion. GTPCH and TH converge biochemically on tyrosine to allow its conversion to L-Dopa, and thus share in this rate-limiting step for catecholamine production. This provides fundamental molecular insights and clinical opportunities. One of the latter is that GTPCH deficiency occurs in humans, causes disease, and can potentially be corrected.

Enzymes in a metabolic pathway are non-rate-limiting when their increase does not increase turnover in the pathway: any enzyme becomes rate-limiting when its decrease or mis-trafficking cause its abundance to be less than the previous ‘rate-limiting enzyme’ in the pathway. Deficiency of AADC causes motor impairment that can be corrected by expression of AADC via viral vector-mediated gene delivery directly to the brain.

Insights into how different types of catecholaminergic neurons develop in the brain have indexed progress in developmental biology as a field. A surprising new insight is that the brain during development may itself be an endocrine organ secreting catecholamines to the rest of the body. The presumption of developmental neuroscientists that ‘TH-positive’ neurons are dopaminergic or noradrenergic has been shattered in the last decade: there are DOPAergic and trace aminergic neurons, as well as neuronal dyads that synthesize dopamine only via intercellular collaboration. The notion that the chemical neuroanatomy of the human nervous system can be fully comprehended by study of the rodent nervous system has likewise been rendered untenable, and translational neuroscience is far better for it. Finally, exciting new vistas have emerged from the use of genetically-based lesioning and complementation experiments that show that subdivisions of the major catecholamine nuclei of the brain, including the locus coeruleus and substantia nigra, have surprising heterogeneity of projections with specific and distinct functions.

Is further progress mainly a matter of effectively executing therapeutic strategies, or do further questions remain? They do. It is still unclear for example how supply of AADC to cells of the striatum corrects a deficiency that inheres primarily in neurons that project to the striatum. The view that imaging of VMAT2 with TBZ reflects the molar concentration of the protein in the brain has given way to an emerging understanding that endogenous catecholamine levels play a role in how much TBZ binds to VMAT2 in the brains of normals, addicts to methamphetamine or cocaine, and patients with progressive degenerative disorders. The near-complete success of virus-mediated correction of brain defects related to catecholamine biosynthesis, and pharmacological correction of genetic deficiencies in catecholamine storage (see for example Rilstone et al., N. Engl. J. Med. 368, 543, 2013) show that robust clinical gains have resulted from translation of pre-2000 information, tools, and advances in allied fields. This in turn predicts that gains in new knowledge reported in this first, as well as the following chapters of Catecholamine Research in the 21st Century will likely parlay into further therapeutic gains in the coming decade.

Genetic Manipulation of Catecholamine Signaling in the Mouse

Richard Palmiter,    University of Washington School of Medicine Seattle, Washington, USA

Gene targeting in mouse embryonic stem cells has been used to generate mice that are unable to synthesize each of the major catecholamines. The developmental and behavioral consequences of making mice that are unable to synthesize epinephrine, norepinephrine and dopamine will be discussed. I will illustrate how restoring dopamine signaling to specific brain regions of an otherwise dopamine-deficient mouse can restore viability and the ability to engage in specific behaviors. I will also show how we manipulate the activity of dopamine-producing neurons to affect the behavior of mice.

AADC Deficiency

Occurring in Humans; Modeled in Rodents; Treated in Patients

Wuh-Liang Hwu¹, Ni-Chung Lee¹, Yih-Dar Shieh¹, Kai-Yuan Tzen², Pin-Wen Chen¹, Shin-ichi Muramatsu³, Hiroshi Ichinose⁴ and Yin-Hsiu Chien¹,    ¹Department of Medical Genetics, Pediatrics,    ²Nuclear Medicine, National Taiwan University Hospital,    ³Division of Neurology, Department of Medicine, Jichi Medical University, Japan;,    ⁴Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Japan

Aromatic L-amino Acid Decarboxylase (AADC) deficiency (MIM #608643) is an autosomal recessive inborn error of neurotransmitter metabolism which causes severe motor dysfunction, oculogyric crisis, autonomic dysfunction, and emotional liability in patients since infancy. AADC deficiency is more common in Taiwan than in other countries because of a foundermutation (IVS6+4A>T). We have established a knock-in (KI) mouse model (DdcIVS6/IVS6) for AADC deficiency. Some of the homozygous KI mice were born alive, but they exhibited severe failure to thrive, dyskinesia, and clasping. However, if they could survive their first few weeks of lives, they then caught up in growth and improved in motor function. Gene therapy at neonatal stage could eliminate the manifestations of the disease. An attempt to treat patients with AADC deficiency was initiated before the establishment of the mouse model. AAV2-AADC vectors were infusued into the bilaterally putamen of four patients 4 to 6 years of age. All of the patients showed improvements in motor performance: one patient was able to stand 16 months after gene transfer, and the other three patients gained head controls or achieved supported sitting. Choreic dyskinesia was observed in all patients, but this resolved after several months. 6-[18F]fluorodopa positron emission tomography (PET) and cerebrospinal fluid analysis both showed evidences of the treatment effects.

Tyrosine Hydrolylase and Dopamine Beta-Hydroxylase

Role of Common Genetic Variation in Adrenergic Responses to Stress and in