Transmissible Spongiform Encephalopathies

Transcription

1 Cambridge Healthtech Institute s 12th Annual Final Agenda REGISTER BY JANUARY 11 th AND SAVE UP TO $200 Transmissible Spongiform Encephalopathies February 11-12, 2008 Sheraton Inner Harbor Hotel Baltimore, MD THE DEFINITIVE AMERICAN TSE MEETING Featuring New Data Emerging Concerns: De novo Formation of Prions Pathogenesis Detection Cell Cultures: From the Bench to the Bed Treatment, Removal, or Inactivation Lead Sponsoring Publications: Hear From These Leading Organizations Rocky Mountain Laboratories - NIH Case Western Reserve University Warwick University Neurological Institute Carlos Besta Amorfix Life Sciences R-Biofarm AG CJD Surveillance Unit - Edinburgh University of Milan University of California - San Francisco University of Maryland and VA Medical Center Istituto Superiore Di Sanita...and many others Cambridge Healthtech Institute, 250 First Avenue, Suite 300, Needham, Massachusetts Telephone: or toll-free in the U.S Fax:

2 February 11, 2008 CHI s Transmissible Spongiform Encephalopathies is the longest running meeting of its kind in the world. This 12 th Annual meeting will address the ongoing progress in the science of prion diseases, as well as the newest developments in the fields of pathophysiology, transmission, detection, removal/inactivation, treatment, and prevention. This conference will present the newest data on TSEs in the context of its application to the pharmaceutical, biological, environmental and device industries. Scientific Advisors: Larisa Cervenakova, M.D., Ph.D., Senior Scientist, Transmissible Diseases Department, Holland Laboratory, American Red Cross Suzette A. Priola, Ph.D., Senior Investigator, Chief, TSE/Prion Molecular Biology Section, Laboratory of Persistent and Viral Diseases, Rocky Mountain Laboratories, NIH Monday, February 11, :00 am Registration and Morning Coffee Emerging Concerns: De novo Formation of Prions 8:00 Welcome by Session Chairperson Paul W. Brown, M.D. 8:15 Ultra-Sensitive Prion Assays Based on Seeded Conversions of Recombinant Prion Protein Byron W. Caughey, Ph.D., Senior Investigator, Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIH PrPSc can seed the conformational conversion and polymerization of normal protease-sensitive prion protein (PrP-sen). Soto and colleagues have shown that this seeding activity allows ultrasensitive detection of prions using cyclical sonicated amplification (PMCA) reactions and brain homogenate as a source of PrP-sen. Building on the PMCA approach, we have developed faster, simpler prion detection methods using recombinant PrP-sen (rprp-sen) which can discriminate normal hamster brain homogenates from scrapie brain homogenates containing <1 intracerebral lethal dose within 2-3 days. In periodically sonicated or shaken cell-free reactions, sub-femptogram equivalents of PrPSc seeded the conversion of rprp-sen into easily detected quantities of specific protease-resistant PrP fibrils. Diseased and normal hamsters were also distinguished using 2-μl of cerebral spinal fluid as seeds. The relative speed, simplicity, replicability and sensitivity of these reactions should facilitate both the development of practical prion assays and structural analyses of prion-seeded PrP polymers. 8:40 De Novo Formation of Purified Native Prions Nathan R. Deleault, Ph.D., Department of Biochemistry, Dartmouth Medical School To study the mechanism of prion formation biochemically, we conducted a series of serial Protein Misfolding Cyclic Amplification (spmca) reactions using purified native PrPC and synthetic polyanionic molecules as substrates. For the first time, we demonstrate that infectious, wild type prions can be: (1) propagated in vitro using purified substrates, and (2) generated de novo from non-infectious components. Furthermore, we have observed that polyanionic molecules are selectively incorporated into physical complexes with PrP during the formation of purified prions in vitro. 9:05 De novo Generation of Prion Infectivity in a Cell-Free System Joaquin Castilla, Ph.D., Assistant Professor, Department of Infectology, Scripps Research Institute-Florida Transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative disorders affecting both humans and animals. There is no available treatment or therapy for these fatal diseases. The infectious agent associated with TSEs (termed prion) appears to be composed uniquely of a protein, which is a conformationally-modified version (PrPSc) of the cellular prion protein (PrPC). The disease is propagated by the conversion of host PrPC into PrPSc induced by small quantities of PrPSc. Interestingly, prions occur in the form of different strains that show distinct biological and physicochemical properties. TSEs can have diverse origins, including genetic, sporadic (putatively spontaneous) and infectious. The occurrence of sporadic cases of prion diseases in humans and maybe in other species, i.e. atypical bovine spongiform encephalopathy (BSE) in European and USA cattle and atypical scrapie cases in sheep suggest that spontaneous prion diseases may happen infrequently but ubiquitously. However, there are no reported cases of spontaneously-occurring prion disease in experimental wild-type rodent models. We have used a novel technique, Protein Misfolding Cyclic Amplification (PMCA) to rapidly propagate prions in the test tube, using normal brain homogenate as substrate. Prions propagated in vitro are infectious in vivo and maintain their prion strain specificity. PMCA has been used to efficiently amplify a variety of prion strains from mouse, hamster, bank vole, deer, cattle, sheep and human. Therefore, to mimic spontaneous generation of infectivity in vitro becomes one of the most important challenges in the prion field. We show here, for the first time, the de novo generation of infectious prions from bank voles (Clethrionomys glareolus) starting with non-infectious brain homogenates. Several biochemically different prion strains were generated using two different wild-type vole genotypes. The de novo in vitro generated PrPSc was highly infectious after its inoculation in bank voles. We show an extensive characterization of this spontaneous phenomenon. 9:30 PMCA Amplification of Prion Amyloid without Amplification of Infectivity Robert G. Rohwer, Ph.D., Director, Molecular Neurovirology Laboratory, Veterans Affairs Medical Center; and Associate Professor of Neurology, School of Medicine, University of Maryland at Baltimore Employing the original protocol for PMCA developed by Soto and colleagues, we obtained a 16 to 32 fold amplification of PK resistant PrP as determined by two fold serial dilution to the starting concentration on Western blot. In comparison, there was no difference in titer, as measured by limiting dilution titration, between the frozen control, a sample that was incubated at 37 C without sonication and the sonicated sample that produced the amyloid amplification. The limiting dilution titration method is sufficiently sensitive to have detected even a 20% difference in titer between the samples. A two fold increase in titer would have caused the infection of nearly every animal at the limiting dilution and could not have been missed. If there is amplification of infectivity during PMCA, it must follow very different kinetics from the amyloid. 9:55 Silent Prions in Normal Brains Wen-Quan Zou, M.D., Ph.D., Assistant Professor, Neuropathology, Case Western Reserve University The co-existence of cellular prion protein (PrPC) and its pathological isoform (PrPSc) is a prerequisite for the pathogenesis of prion diseases. However, molecular mechanism of PrPSc formation in the spontaneous prion diseases including sporadic and familial forms remains poorly understood. Our recent studies indicate that in the uninfected brain there are small amounts of abnormal PrP species that may be involved in the pathogenesis of spontaneous prion diseases. 2

3 10:20 Discussion with all Session Speakers 10:40 Coffee Break, Poster and Exhibit Viewing Pathogenesis Chairperson: Larisa Cervenakova, M.D., Ph.D. 11:10 Accumulation of Prion Protein in the Brain That is Not Associated with Transmissible Disease Pedro Piccardo, M.D., Senior Investigator, OBRR / DETTD / LBPUA, FDA 11:35 High Levels of TSE Infectivity Can Be Associated with Little or No Detectable PrPSc in Vivo Rona Barron, Ph.D., Neuropathogenesis Unit, Roslin Institute and Royal (Dick) School of Veterinary Studies This work examines the relationship between TSE infectivity and the abnormal prion protein, PrPSc. In a mouse model of disease we have shown high titres of TSE infectivity in brain tissue which contains little or no PrP-res. We also found no evidence of other abnormal PrP isofoms such as PK-sen PrPSc. These data question the true relationship between PrPSc and TSE infectivity, and the current reliance on PrPSc as the sole diagnostic marker for TSE disease. 12:00 Conversion of the BASE Prion into the BSE Prion: The Origin of BSE? Fabrizio Tagliavini, Ph.D., Director, Division of Neurology 5 & Neuropathology, Neurological Institute Carlo Besta Twenty years after the identification of bovine spongiform encephalopathy (BSE), the origin of the causal agent is still unknown. This issue is of fundamental importance, since knowledge of the origin of the BSE agent is essential for prevention of future outbreak of the disease or variants thereof in cattle and other mammals. We carried out transmission studies with transgenic mice expressing bovine PrP and four lines of non-transgenic mice and found that an atypical form of spongiform encephalopathy of cattle, termed BASE or BSE-L, is caused by a prion strain distinct from that of classical BSE. Noteworthy, this newly characterized prion strain has the ability to convert into the classical BSE strain upon serial transmission to inbred mouse lines. According to these results, BASE--which is regarded as a sporadic form of prion disease in cattle--may be the origin of BSE, following conversion of the causal agent in an intermediate host. 12:45 Luncheon Technology Workshop (Sponsorship Available) or Lunch on Your Own 2:00 Sporadic CJD and Atypical BSE: Two Children of One Protein Maurizio Pocchiari, M.D., Director of Research, Virology, Istituto Superiore Di Sanita The identification of forms of TSE diseases in cattle caused by prion strains different from BSE has raised new concerns on the possibility that these novel agents might induce disease in humans with a phenotype resembling sporadic CJD. The analysis of the distribution of the different molecular subtypes of sporadic CJD might give some answers. 2:25 Variant CJD: Residual Uncertainties Robert Will, M.D., National CJD Surveillance Unit, Edinburgh, UK Mortality from variant CJD continues to decline, but concerns for public health persist. These are based on uncertainty on the population prevalence of infection, the incubation period of vcjd and the potential for further cases of secondary transmission. Information from epidemiology, molecular biology and transmission studies may provide new insights into these issues. 2:50 Panel Discussion Detection 3:05 Comments by Session Chairperson Byron W. Caughey, Ph.D., Senior Investigator, Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIH 3:10 Biochemical Detection of Prions in Blood and Urine Claudio Soto, Ph.D., Professor, Neurology, University of Texas-Galveston 3:35 The Elusive Precursor of PrPSc Teresa Pinheiro, Ph.D., Professor, Biological Sciences, University of Warwick A key molecular event in prion diseases is the conversion of the cellular prion protein, PrPC, to the aberrant disease-associated state, PrPSc. The details of this molecular transformation are not fully understood, but it has been suggested that an intermediate on the normal folding pathway of PrPC may be recruited to form PrPSc. Using conventional analyses of folding transition data determined by fluorescence and circular dichroism, and novel phase diagram analyses, we present compelling evidence for the presence of an intermediate species on the folding pathway of PrP. This state is structurally close to the native state and upon incubation produces off-pathway aggregates. The results explain the difficulties in prion diagnosis and suggest possible new directions for test development. 4:00 Afternoon Refreshment Break, Poster and Exhibit Viewing 4:30 Detection of Blood Prions with Epitope Protection Technology Neil Cashman, M.D., Professor, Medicine, Amorfix Life Sciences In order to detect prions, an assay must distinguish between the normally folded prion protein PrPC and its aggregated disease-causing conformation PrPSc. We have developed a chemical means to differentiate between PrPC and PrPSc, called Epitope Protection technology. PrPC is selectively modified with short-lived and highly-reactive chemicals which modify selected amino acids in the protein. Such chemical modification efficiently blocks immunological epitopes on PrPC and leaves them unrecognizable to many PrP antibodies. PrP molecules within prion particles are protected from chemical modification, and can then be detected by conventional immunoassay after disaggregation of the sample. We now report detection of the very small concentration of PrPSc, which has been estimated in the low femtogram range per ml of infected blood. We have tested this methodology by screening blinded panels of vcjd brain and spleen material spiked into plasma, and we can reliably detect vcjd brain homogenate after diluting a 10% homogenate more than 105-fold into plasma. A high-throughput version of the assay has been developed and allows the processing of thousands of samples per day. 4:55 PrionScreen - All TSEs on a Plate Martin Mehl, Ph.D., Product Manager BSE, Marketing, R-Biopharm AG PrionScreen, the Roche Diagnostics last generation ELISA kit, provides the actual demands to identify all relevant typical and atypical TSEs from various species within the same assay. The overall changes in test kit requirements and the data of the recent EU approvals are summarized from the industries view. 5:20 Discussion with all Session Speakers 5:35 Networking Reception in the Exhibit Hall 6:40 Close of Day One 3

4 February 12, 2008 Tuesday, February 12, :30 am Morning Coffee Cell Cultures: From the Bench to the Bed 9:00 Comments by Session Chairperson Suzette Priola, Ph.D 9:05 Exploring Pathogenesis in Cell Culture Suzette A. Priola, Ph.D., Senior Investigator, Chief, TSE/Prion Molecular Biology Section, Laboratory of Persistent and Viral Diseases, Rocky Mountain Laboratories, NIH The early events that occur during TSE infection are largely unknown. Using a tissue culture system that enables us to exclusively track abnormal prion protein (PrPSc) during acute exposure of cells to TSE infectivity, we have studied the initial interaction between PrPSc and the cell. Our data suggest that early events during TSE infection may be largely strain and cell-type independent. Understanding the acute stages of TSE infection may provide new targets and strategies for TSE prevention and/or therapeutics as well as advance our knowledge of basic TSE pathogenesis. 9:30 Talk Title to Be Announced Larisa Cervenakova, M.D., Ph.D., Senior Scientist, Transmissible Diseases Department, Holland Laboratory, American Red Cross 9:55 Clearance and Prevention of Prion Infection in Cell Culture by Anti-PrP Antibodies Thomas M. Wisniewski, M.D., Professor, Neurology, Pathology and Psychiatry, New York University School of Medicine 10:20 Discussion with All Session Speakers 10:35 Coffee Break, Poster and Exhibit Viewing Treatment, Removal, or Inactivation Chairperson: Robert G. Rohwer, Ph.D., Director, Molecular Neurovirology Laboratory, Veterans Affairs Medical Center; and Associate Professor of Neurology, School of Medicine, University of Maryland at Baltimore 11:05 Anti-TSE Immunization in a Hamster Model Giorgio Poli, Professor, DVM, Veterinary Pathology, University of Milan Vaccination has been shown to be effective in mouse models for neurodegenerative conditions characterized by protein misfolding, such as Alzheimer s disease (AD) and Transmissible Spongiform Encephalopathies (TSEs) and many different immunogens and strategies of intervention have been proposed. Here we show that the immunization of hamster with a synthetic oligopeptides PrP , corresponding to the central part of hamster prion protein, prolonged the survival time (>23 days) in animals challenged with the 263K strain of scrapie agent. The immunized hamster mounted a specific, but weak, antibody response, and developed also a specific cellmediated response (as shown by proliferation assay on splenocytes). Moreover, the results obtained with RT-Real Time PCR showed an over-expression of mrna for GFAP and pro-inflammatory cytokines (TNF-a, IL-1β) in brain of infected controls compared to immunized animals and healthy controls, as well as an decrease in IL-10 expression. Increased survival after challenge was also associated with reduction of cerebral lesions, glial fibrillary acid protein (GFAP) and PrP deposition (both in brain and spleen). Our results indicate that a humoral and cell mediated immune response can slow down PrPres deposition and decrease neuroinflammation; nevertheless, it is conceivable that the vaccination can activate in peripheral organs specific different and interacting mechanisms, as observed in other scrapie mouse models. 11:30 Application of PRDT Affinity Binding Technology to a Reduction of Risk of TSE Transmission in the Blood and Biopharmaceutical Industry Luisa Gregori, Ph.D., Assistant Director MNV Laboratory and Assistant Professor University of Maryland, BREF, University of Maryland and VA Medical Center Transmissible spongiform encephalopathy (TSE) diseases are neurodegenerative illnesses that can be transmitted by blood transfusion. Precautionary measures against the spread of TSE through the blood supply have been implemented, but alone those measures are not sufficient. Pathogen Removal and Diagnostic Technologies, Inc. (PRDT) developed a new strategy based on the use of ligands derived from a combinatorial chemistry approach that specifically adsorb the TSE agent. After screening several million compounds, PRDT identified a ligand that when coupled in resin format reduced the TSE causative agent from blood to the limit of detection of a bioassay. This resin was incorporated into a prion reduction filter, the P-CAPTTM filter, manufactured by MacoPharma. The product has already demonstrated utility in reducing infectivity > 3log 10 (brain derived infectivity in red blood cell concentrate - RBC) and > 1.2log 10 (endogenous whole blood derived infectivity). 11:55 Differential Resistance of Prions Strains to Inactivation Kurt Giles, Ph.D., Assistant Adjunct Professor, Institute for Neurodegenerative Diseases, University of California San Francisco Understanding prion inactivation is essential for avoiding iatrogenic transmission of CJD via surgical instruments, and ensuring the safety of the food supply. Using a range of transgenic mouse models sensitive to naturally occurring and rodent-adapted prion strains, we have determined the difference in resistance to inactivation for a range of prion strains. By applying a set of rigorous statistical approaches we have produced the most comprehensive study of prion inactivation. 12:20 Closing Comments by Scientific Advisors 12:30 Close of Conference 4

5 Cambridge Healthtech Institute s 12th Annual Transmissible Spongiform Encephalopathies THE DEFINITIVE AMERICAN TSE MEETING Sponsoring Publications: Web Partners: Sponsor and Exhibitor Information Showcase your company s expertise, brand your solutions and develop revenue-generating opportunities with qualified decision-makers by Exhibiting or Sponsoring. Sponsorship packages are designed to achieve your business development and networking goals and objectives. Sponsorship benefits include pre-conference, at-conference and post-conference marketing efforts. The earlier you secure your sponsorship, the more opportunity for exposure. Numerous promotional and sponsorship programs exist and are flexible to allow any size company to participate. Sponsored speaking opportunities may include: Technology Spotlight, embedded within the conference program Breakfast or Luncheon Workshops, allow for podium time Embedded Presentations Other sponsorship opportunities include sponsoring an Invite-Only VIP dinner to access this audience at the highest level, and ensure optimum face-to-face networking. Hotel Information Sheraton Inner Harbor Hotel 300 S. Charles Street Baltimore, MD Tel: Fax: Room Rate: $169 s/d Reservation Cutoff: January 11, 2008 Car Rental Information Special discount rentals have been established with AVIS for this conference. Please call AVIS directly at and you must reference your Avis Worldwide Discount (AWD) Number J For more information, to reserve booth space, or discuss sponsorships contact: Katelin Martin, kmartin@healthtech.com Call for Posters Reasons You Should Present Your Research Poster at Transmissible Spongiform Encephalopathies: Your poster will be exposed to our international delegation Receive $50 off your registration Your poster abstract will be published in our conference CD Your research will be seen by leaders from top pharmaceutical, biotech, academic and government institutes 5 Photo Credit: Dr. Al Jenny, 2003 (Content Providers(s): U.S. Dept. of Agriculture - Animal and Plant Health Inspection Service, APHIS)

6 Cambridge Healthtech Institute s 12th Annual Transmissible Spongiform Encephalopathies February 11-12, 2008 Sheraton Inner Harbor Hotel Baltimore, MD YES! Register me for Transmissible Spongiform Encephalopathies REGISTRATION INFORMATION Mr. Ms. Mrs. Dr. Prof. Name Job Title Div./Dept. Company Address City/State/Postal Code Country Telephone Would you like to receive event updates via fax? Yes No Fax * * is not a mandatory field. However, by excluding your you will not receive notification about online access to pre-conference presenter materials, conference updates and networking opportunities. PRICING INFORMATION COMMERCIAL ACADEMIC, GOVERNMENT, HOSPITAL AFFILIATED TWO DAY CONFERENCE REGISTER 3 4 th IS FREE Individuals must register for the same conference or conference combination and submit completed registration forms together for discount to apply. Please reproduce this registration form as needed Advanced Registration until January 11, 2008 $1195 $620 Registration after January 11, 2008 or On-Site $1395 $695 Poster Discount $50 Off $50 Off I cannot attend but would like to purchase the Transmissible Spongiform Encephalopathies conference CD for $250 (plus shipping). Massachusetts delivery will include 5% sales tax. Please send information on exhibiting and opportunities to present workshops. PAYMENT INFORMATION Enclosed is a check or money order payable to Cambridge Healthtech Institute, drawn on a U.S. bank, in U.S. currency. Invoice me, but reserve my space with credit card information listed below. Invoices unpaid two weeks prior to conference will be billed to credit card at full registration rate. Invoices must be paid in full and checks received by the deadline date to retain registration discount. If you plan to register on site, please check with CHI beforehand for space availability. Please charge: AMEX (15 digits) Visa (13-16 digits) MasterCard (16 digits) Diners Club (14 digits) Card # Exp. Date Cardholder Signature Cardholder s Address (if different from above) City/State/Postal Code Country Please refer to the Keycode below: Yes! I would like to receive a FREE subscription to: a monthly magazine focused on technologies for the life sciences the premiere e-news source on technology for healthcare a weekly e-newsletter report on the technologies advancing clinical trials 838 F Present a Poster and Save $50! Cambridge Healthtech Institute encourages attendees to gain further exposure by presenting their work in the poster sessions. To secure a poster board and inclusion in the conference CD, your abstract must be submitted, accepted and registration paid in full by January 23, Register online to use the Poster Abstract Submission form or, if you register by phone, fax, or mail, you will receive Poster Abstract Submission guidelines via . I am interested in presenting a poster at Transmissible Spongiform Encephalopathies and will submit a completed one-page abstract by January 23, 2008 (Please Note: Registration must be paid in full to present poster.) Title CHI insight Pharma Reports A series of reports that evaluate the salient trends in pharmaceutical technology, business, and therapy markets. Keep abreast of the latest advances in pharmaceutical R&D, their potential applications and business impacts, and their current and future position in the marketplace. For a list of reports, visit Insightpharma.com, or contact Rose LaRaia, rlaraia@healthech.com, Additional Registration Details Each registration includes all conference sessions, posters and exhibits, food functions, and a copy of the conference CD. Group Discounts Special rates are available for multiple attendees from the same organization. Contact David Cunningham at to discuss your options and take advantage of the savings. Handicapped Equal Access In accordance with the ADA, Cambridge Healthtech Institute is pleased to arrange special accommodations for attendees with special needs. All requests for such assistance must be submitted in writing to CHI at least 30 days prior to the start of the meeting. Substitution/Cancellation Policy In the event that you need to cancel a registration, you may: Transfer your registration to a colleague within your organization Credit your registration to another Cambridge Healthtech Institute program Request a refund minus a $100 processing fee per conference Request a refund minus the cost ($250) of ordering a copy of the CD NOTE: Cancellations will only be accepted up to two weeks prior to the conference. Program and speakers are subject to change. Video and or audio recording of any kind is prohibited onsite at all CHI events. Cambridge Healthtech Institute 250 First Avenue, Suite 300, Needham, Massachusetts T: or toll-free in the U.S F: