Scientist Solutions - Protein Folding Disorders

Day 1

Alzheimers, Parkinson's Disease, one of the inherited forms of emphysema, CJD, Cystic Fibrosis, Type 2 Diabetes, and even some cancers are all related to the abnormal folding of proteins within the body. These devastating diseases cause untold misery, extracting both monetary and human tolls. This highly focused conference will discuss strategies for elucidating protein misfolding mechanisms, methods for improved study of these events and applications to the development of effective drug discovery and development for the treatment of these disorders. Researchers doing work in any related research areas are encouraged to participate.

Scientific Advisors
Dr. Steve Abcouwer, Penn State University-Hershey Medical Center
Dr. Susan M. Catalano, Acumen Pharmaceuticals, Inc.

Transmissible Spongiform Encephalopathies and Protein Folding Disorders
Joint Session
10th International
Transmissible Spongiform Encephalopathies
Transmissible Spongiform Encephalopathies
The Definitive American TSE Meeting
March 7-9, 2006 2nd International
Protein Folding Disorders
March 9-10, 2006

Thursday, March 9

8:30 Morning Coffee

Therapies

9:00 Comments by Session Chairperson
Dr. Paul Brown

9:15 Vaccine Approaches to Prevent and Treat Prion Infection
Professor Thomas Wisniewski, Professor, Neurology, Pathology and Psychiatry, NYU School of Medicine
Immunological therapeutic approaches are currently being developed for a number of conformational neurodegenerative conditions, including Alzheimer's disease (AD) and prion disease. Passive immunization is now in clinical trial for the treatment of AD. We were the first to show in wild-type prion model mice that active and passive immunization can produce resistance to prion infection with prolongation of the incubation period. We are currently developing mucosal prion vaccines which can prevent infection in a proportion of animals later exposed to the prion agent orally. These studies can be used to develop active vaccines for livestock and eventually in humans who are at risk for developing prion infection.

9:45 Searching for Anti-TSE Compounds using Animal Models
Dr. David Kocisko, Staff Scientist, NIH/Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories
There is no treatment for the TSEs and they are always fatal. A number of potential TSE therapies were tested in transgenic mice and several new compounds have been found with in vivo prophylaxis activity. One of these new compounds also has activity when used as a treatment following scrapie inoculation into the brain, which places it in a small group of molecules with such activity.

10:15 Protein Origami: Molecular Folding in Health and Disease
Dr. P. Michael Conn, Associate Director and Senior Scientist, The Oregon National Primate Research Center; Special Assistant to the President and Professor of Physiology, Pharmacology, and Cell and Developmental Biology, Oregon Health and Science University
Mutants of the gonadotropin releasing hormone receptor are frequently misrouted within the cell, but otherwise competent proteins. "Pharmacoperones," small molecules that penetrate cells and correct folding, rescue such
"defective" molecules, allowing the misfolded mutants to escape retention by the cell's quality control apparatus and route correctly to the plasma membrane. This observation and the indication that disease is a frequent corollary of protein misfolding/misrouting suggests that protein rescue may be an under-appreciated therapeutic approach. The observation that a percentage of wild-type proteins may also be misrouted suggests that this is a novel form of post-translational regulation associated with normal function that can also be therapeutically exploited.

10:45 Technology Short Talk (Sponsorship available)

11:00 Coffee Break, Poster and Exhibit Viewing

SCIENCE AND POLICY: Joint FDA/USDA Panel on TSE-Safe Products: The Federal Role

11:30 Comments by Session Chairperson
Dr. Kiki Hellman, Hellman Associates LLC

11:35 Reducing TSE Risk for Biological Products: Regulatory Approaches
Dr. David M. Asher, Chief, Laboratory of Bacterial, Parasitic and Unconventional Agents, Division of Emerging and Transfusion-Transmitted Diseases, Office of Blood Research and Review, CBER, Food and Drug Administration

11:50 TSE Transmission by Surgical Instruments: Evaluation of Products Proposing to Reduce that Risk
Dr. Sheila A. Murphey, Branch Chief, Infection Control Devices Branch, Division of Anesthesiology, General Hospital, Infection Control and Dental Devices, Office of Device Evaluation, Center for Devices and Radiologic Health, Food and Drug Administration

12:05 Reducing TSE Risk for Biotechnology Products: Regulatory Approaches
Dr. Gerald Feldman, Senior Investigator, Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS Center for Drug Evaluation and Research, Food and Drug Administration

12:20 CVM Update on Feed Controls for Prevention of BSE
Dr. Burt Prichett, Division of Animal Feeds, Center for Veterinary Medicine, Food and Drug Administration

12:35 Title to be announced
Dr. Lisa Ferguson, Senior Staff Veterinarian, United States Department of Agriculture, APHIS, Veterinary Services

12:50 Lunch and Learn Workshop
(Sponsorship Available)

Close of Transmissible Spongiform Encephalopathies

Continuation of
2nd International
Protein Folding Disorders
March 9-10, 2006

Thursday, March 9

ADVANCES IN THE SCIENCE

2:10 Comments by Session Chairperson
Dr. Susan Catalano, Acumen Pharmaceuticals

2:15 Do Amyloid Channels Cause Disease?
Dr. Bruce Kagan, Professor, Psychiatry, UCLA
At least nine different amyloid peptides form non-selective ion-permeable channels in membranes under conditions leading to oligomerization and cytotoxicity. We propose that beta sheet structure leads naturally to membrane channel formation, and that these channels have properties that would lead to cellular dysfunction and death. The striking similarity of the channel properties of these diverse peptides suggests they possess a common structure that might be a target for therapeutic drug development. Accumulating evidence that these channels form in membranes in vivo, implies that they may be inaccessible to anti-aggregation agents and may require targeting with channel blocking agents.

2:45 VCP/p97 is Involved in Endoplasmic Reticulum Associated Degradation of CFTR: Implications in Cystic Fibrosis
Dr. Neeraj Vij, Pediatric, Pulmonary, Johns Hopkins University
Cystic fibrosis results when functional levels of CFTR are reduced at the cell surface. Both the most common mutant ΔF508 and wild type- CFTRs are targeted for ERAD. We show here that p97/VCP and gp78 form complexes with CFTR during translocation to the proteasome. Interference in the VCP/CFTR binding promotes accumulation of immature CFTR in the ER. Inhibition of the proteasome with PS-341 (Bortezomib/Velcade) rescues CFTR and leads to appearance of mature CFTR at the cell surface. Moreover, under these conditions, IL8 cytokine levels were reduced, suggesting that exaggerated inflammatory phenotype in CF is casually linked to proteasome activity.

3:15 N-Methylated Peptide Inhibitors of Beta-Amyloid Aggregation
Professor Andrew Doig, Professor, Faculty of Life Sciences, University of Manchester
N-methylated peptide fragments and derivatives can act as inhibitors of amyloidogenic proteins and peptides. We optimized N-methylated peptide structures to inhibit beta-amyloid aggregation by screening a number of libraries. The extent of amyloid formation was quantified primarily by Thioflavin T binding and electron microscopy, while beta-amyloid toxicity was measured by several assays on PC12 or SHSY5Y cells. We found more potent compounds than all other known peptides, peptidomimetics and small molecule inhibitors. Optimized inhibitors were able to reverse the toxic effects of beta-amyloid at nanomolar concentration on an in vitro brain slice, determined by long term potentiation.

TECHNOLOGY SOLUTIONS
3:45 Technology Workshop Title and Speaker To Be Announced

Sponsored by

4:00 Afternoon Refreshment Break, Poster and Exhibit Viewing

SCREENING & ASSAYS

4:30 Molecular Chaperones are Required for the Mobility of Nuclear Receptors in Living Cells
Dr. Gordon Hager, Lab Chief, Receptor Biology and Gene Expression, National Cancer Institute
The development of green fluorescent protein (GFP) technology combined with live cell microscopy techniques revealed the dynamic properties of proteins in the nucleus. FRAP (fluorescence recovery after photobleaching) experiments demonstrate that many nuclear proteins are highly mobile within the nucleus. We have developed a novel in situ assay that combines a biochemical permeabilization and extraction procedure with a quantitative FRAP technique. Using this assay, we have found a new functional role for molecular chaperones in the nuclear mobility of steroid receptors.

5:00 RNAi Screening to Identify Genetic Factors Associated With Parkinson's Disease
Professor Guy Caldwell, Associate Professor, Biological Sciences, The University of Alabama
We have exploited the advantages of the transparent nematode, C. elegans, to systematically screen for factors that affect susceptibility to Parkinson's Disease (PD) by investigating genes that modulate alpha-synuclein misfolding in vivo. We have employed RNAi to conduct a large-scale screen for genetic factors influencing the misfolding of a human alpha-synuclein::GFP fusion as worms age. From over 900 total targets, we have identified 18 positive genes that result in increased alpha-synuclein misfolding. As a secondary test for any putative relationships to PD, we have thus far shown 2 of these alpha-synuclein effectors can protect C. elegans dopaminergic neurons from oxidative damage and neurodegeneration when their corresponding cDNAs are over expressed in transgenic nematodes. This data provides a paradigm for the investigation of cellular factors that influence the management of protein misfolding and its consequences for movement disorders.

5:30 Close of Day

Friday, March 10

8:30 Continental Breakfast in Exhibit Hall

APPLICATIONS

9:00 Cooments by Session Chairperson
Stephen Abcouwer, University of Pennsylvania-Hershey Medical Center

9:15 Understanding and Preventing the Initial Events in Amyloid B-Protein Assembly - Towards Therapeutics for Alzheimer's Disease
Dr. Gal Bitan, Assistant Professor in Residence, Department of Neurology, David Geffen School of Medicine, University of California at Los Angeles
Soluble, pre-fibrillar oligomers of the amyloid b-protein (Ab) are believed to be the main neurotoxins acting in Alzheimer's disease (AD). The harmful action of these oligomers leads to synapse dysfunction, neuronal loss, dementia, and eventually death. Inhibition of Ab oligomerization, therefore, is an attractive strategy for prevention and treatment of AD. In order to effectively inhibit Ab oligomerization, the structures and mechanisms of formation of the oligomers must be understood. Studies of these structures and mechanisms are difficult because Ab oligomers possess highly unfavorable characteristics. The oligomers are metastable, exist as rapidly changing mixtures, their solubility in aqueous solutions is poor, and they adhere non-specifically to various substances, matrices, and surfaces. The presentation will discuss the synergistic efforts of several laboratories over the last few years using biological, biochemical, biophysical, and computational methods, which have advanced our understanding of the early events in Ab assembly, and the first attempts to design inhibitors of Ab oligomerization.

9:45 Effects of Intracellular Soluble Misfolded A - Targets of Opportunity
Dr. Harry LeVine, III, Sanders-Brown Center on Aging, University of Kentucky
Recent evidence suggests that histologically invisible oligomeric forms of A elicit potent neurotoxic effects. While it is clear that extracellular oligomers have receptor- and non-receptor-mediated effects on nervous tissue, intracellular oligomers may be plying a more insidious trade. Cells of organisms from bacteria to humans possess quality control mechanisms to prevent the aggregation of misfolded proteins resulting from biochemical stressors or synthesis errors. A network of chaperone/co-chaperone proteins unfold misfolded proteins and then either refold them or target them for degradation. In humans this system and ancillary processes successfully protect the brain against the accumulation of misfolded proteins for 50+ years. Then, in an unexplained transition, A brain levels rise and an increasing proportion of people begin developing brain pathology that eventually manifests as dementia in Alzheimer's disease, movement disorders in Parkinson's disease, and other chronic neurological dysfunction. We find that soluble oligomeric, but not monomeric, A(1-42) associates with the hsp70 chaperone and the BAG-1 co-chaperone in vitro and in cellular extracts. Hsp70 affinity for BAG-1 is increased. A perturbation of the hsp70 regulatory cycle by increasing BAG-1:hsp70 complexes has the potential to alter not only the folding and targeting activities of hsp70, but to chronically disturb the multiple signals intersecting at BAG-1 that balance apoptosis and neuronal differentiation leading to neuronal dysfunction.

10:15 Soluble Amyloid Oligomer Inhibitors for Treatment of Memory-Related Disorders
Dr. Susan Catalano, Director of Discovery Biology, Acumen Pharmaceuticals, Inc.
Acumen Pharmaceuticals focuses on the development of small molecule therapeutics and diagnostics targeting soluble oligomers of amyloid beta 1-42 in Alzheimer's disease and other memory-related disorders. This presentation will briefly review the scientific background implicating these soluble oligomers as causative agents in memory disorders, and discuss the current progress towards identifying small molecule inhibitors of amyloid beta 1-42 assembly, receptor binding and signaling.

10:45 Coffee Break, Poster and Exhibit Viewing

11:15 Hsp90 Inhibitors: A Therapeutic Approach for Tauopathies?
Dr. Leonard Petrucelli, Mayo Clinic
Neurofibrillary tangles (NFT) are a characteristic neuropathological feature of Alzheimer's disease (AD) and molecular chaperones appear to be involved in the removal of disease-associated hyper-phosphorylated tau, a primary component of NFTs. Using a unique quantitiative assay, we have identified novel HSP90 inhibitors which selectively reduced levels of tau phosphorylated at proline-directed Ser/Thr sites (pS202/T205, pS396/S404) and conformationally altered (MC-1) tau species. We will discuss the in vivo implications of Hsp90 Inhibitors for the treatment of tauopathies.

11:45 Small Molecule Chaperone Treatment Restores Enzyme Activity and Protein Trafficking of Mutant Enzymes in Animal Models and Fibroblasts from Patients with Lysosomal Storage Disorders
Dr. Brandon Wustman, Senior Scientist II, Cell Biology, Amicus Therapeutics
Lysosomal storage diseases are historically described as diseases of substrate accumulation. However, mounting data suggest that protein accumulation may lead to secondary pathologies such as Parkinson's and other diseases of aggregation. To address both loss of function and proteotoxicity, we have identified small molecule compounds that may reduce toxic protein accumulation by restoring protein trafficking in cell-based and animal models for Fabry, Gaucher and Pompe diseases.

12:15 Close of Conference

For more information please contact:
Elizabeth Lamb, Conference Director, Cambridge Healthtech Institute
Fax: : 207-493-4573 E-mail: elamb@healthtech.com

For sponsorship or exhibiting information, please contact:
Suzanne Carroll, Manager, Business Development
Phone: 617-630-1352, E-mail: scarroll@healthtech.com