Invasive Aspergillosis Animal Models (IAAM)

Transcription

1 RFA NIH-NIAID-DMID-03-09: New Animal Models for: Part B Invasive Aspergillosis (August 12, 2002) NIH-NIAID-N01-AI Invasive Aspergillosis Animal Models (IAAM) (August 15, 2003-August 14, 2010) I think.therefore IAAM. DMD Sept 03

2 Need for Reference Standard(s) Historical perspective: Critical needs in antifungal susceptibility Method reproducible Provides reference standard for comparison of other methods Validated for clinical correlation Highest priority for Aspergillus and new diagnostics Standardized models aimed at identifying new targets for diagnosis & monitoring of disease progression Establish Methodology Intra-laboratory Variability Inter-laboratory Variability Validation Clinical Correlation

3 SPECIFIC AIMS Establish and standardize animal models of IA Mouse Model Guinea pig Model Rabbit Model Develop molecular tools to provide standardized procedures for genetic manipulation of Aspergillus strains. Molecular Toolbox Pathogenesis Toolbox Determine gene expression of both whole cell and individual target genes of Aspergillus in experimental infection Post-genomic data to develop novel diagnostic and immunologic approaches in the management of IA Dissemination of knowledge and skills to qualified scientists and laboratories Training Website Annual workshops

4 NIAID/DMID Project Officer Rory Duncan Dennis Dixon IA Animal Models (IAAM) Principal Investigator Tom Patterson Steering Committee Denning, Filler, May, Nierman, Walsh Reviews Guides Central Unit Administrative Core: Rick Kirkpatrick/Connie LaBeaux Expert Panel A. Casadevall (Chair) J. Rhodes Functional Components (Working Groups) Patterson, Filler, Sheppard, Denning, Wickes, Pollock Proposes Develops Implements Delivers

5 Animal Models: Key Features Mice & larger animals: rabbit/guinea pigs Pulmonary (aerosol) challenge Neutropenia/nonneutropenic; other immunosuppression Differentiate: exposure/colonization/ infection Standardized Telemetry/IR fever curves Continuous blood sampling for surrogate markers Quantify tissue burden (2 methods) Local/disseminated infection Genomic approach to molecular diagnosis Survival duration allowing for disease progression (4-7 d) Assess growth dynamics of fungi A. fumigatus (AF293); suitable for others Gene profiling

6 Animal Model Design Recapitulates human disease Cost Reproducibility Ease of use Amenable to studies including: Evaluation of novel diagnostics Evaluation of host response Evaluation of organism virulence factors on diagnosis through molecular manipulations In vivo expression analysis Evaluation of effect of therapy on diagnosis

7 Madison Aerosol Challenge/Inhalational Chamber The Reference Standard Precision delivery to: - 90 mice -18 guinea pigs - 4 rabbits

8 Developmental Status Report Completion of Phase I continuation of phase 2 (end of year 4) Deliverables: Standard operating procedures - Molecular techniques - Animal models - How to identify key questions/new diagnostics - Interactions with Steering Committee, Scientific Community & Commercial Partners Procedures for receiving requests and establishing priorities Development of new diagnostics - New targets - New platforms (Provide tissues/samples; testing)

9 IAAM: Standard Operating Procedures

10 Key Questions Accept/ Modify/ Reject Qualified investigator/ Key question Steering Committee Expert Panel IAAM Project Officer (Rory Duncan) IAAM Principal Investigator (Thomas Patterson) Accept/ Modify/ Reject IAAM Executive Committee (Patterson, Filler, Denning)

11 Key Questions Qualified investigator Researcher with interest in Aspergillus Trained to safely perform requested research Qualifications: Principal investigators; Trainees (Post-doctoral fellows, students); Industry researchers Key Questions Gene/gene product as diagnostic target Evaluation of surrogate marker(s) Effects of therapy on disease progression/gene expression Role of virulence determinants in diagnosis Others

12 Key Questions Prioritization of key questions High likelihood of commercialized diagnostic product Data to support development of diagnostic product Pilot studies to test theoretical diagnostic target - NIH funded research - Preliminary data to support NIH application with favorable priority score on review - Pilot studies to evaluate investigator initiated concept - Industry sponsored research

13 Key Questions W. Steinbach R. Cramer B. Miller N. Wiederhold C. Clancy S. Harris R. Akins/J. Sobel A. Zaas R. Calderone M. Momany B. Segal D. Perlin C. Selitrennikoff C. Douglas J. Loeffler G. Ramage Calcineurin pathway in IA Role of glip in gliotoxin synthesis Aspergillus virulence determinants Genome-wide expression to echinocandins for Af In vivo gene expression of Af Polarized Hyphal Growth in Af Microfluidic device for rapid pathogenic fungal diagnosis Genetic determinants of Af susceptibility Germination in Af Rho GTPases in polar growth of Af Development of Aspergillus vaccine New Diagnostics for Af Prophylactic and therapeutic Aspergillus vaccines QPCR for diagnostics of A. fumigatus QPCR for diagnostics of A. fumigatus Real-Time PCR assay to detect A. fumigatus Duke Duke U Idaho UTHSCSA U Florida U Nebraska U Michigan Duke Georgetown U Georgia SUNY/Buffalo New Jersey MycoLogics, Inc, Aurora, CO Merck and Co., Inc, Rahway, NJ University of Wuerzburg, Germany Glasgow Caledonian University, Glasgow, Scotland

14 Key Questions C. Cray D. Sheppard B. Wickes S. Filler / D. Sheppard S. Baker R. Cramer N. Wiederhold N. Wiederhold A. Vallor S. Filler / D. Sheppard M. Del Poeta Russ Lewis Terry Sweeny Murine model of Pulmonary invasive Aspergillosis GM diagnostics in A. fumigatus PCR diagnostics in A. fumigatus Host response to invasive aspergillosis Proteomics approach to A. fumigatus detection Metabolomics approach to A. fumigatus detection Chitin assay development for pulmonary aspergillosis effect of paradoxical effect on diagnosis of IPA during echinocandin therapy utility of serum vs whole blood for assessment of fungal burden in IPA Effect of different aspergillus isolates on experimental murine IPA Detection of anti-glucosylceramide antibody in an Invasive Aspergillosis Animal models for diagnosis and treatment (Use of SOPs) ABIP in an inhalational model of aspergillosis (Use of SOPs) University of Miami Miller School of Medicine, Miami FL McGill University, Montreal Canada UTHSCSA UCLA - Harbor / McGill University Pacific Northwest National laboratory, Richland, WA Duke UTHSCSA UTHSCSA UTHSCSA UCLA - Harbor / McGill University Medical Univ. Of S. Carolina, Charleston, SC MD Anderson, Houston TX Nektar Therapeutics, San Carlos, CA

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16 Future Directions Deliverables/Model refinement SOPs online Alternative Af strains Role of host responses Distinction of colonization vs disease Impact of sample types, collection, storage New target identification Gene product(s) in disease Host gene responses Novel approaches Diagnostic development Industry partners Pre-clinical support for diagnostics Community awareness/interaction

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