RESEARCH & DEVELOPMENT PROJECTS JULY 1, 2001 – JUNE 30, 2004

RESEARCH PROJECTS

Project Title: Activity of ketolides vs. Canadian Gp A Streptococci

Co-PI: George Zhanel, Daryl Hoban, Stephen Douthwaite

Study description: Study assessing the activity of ketolides vs. Canadian Gp A Streptococci

Duration: 2003-2005

Funding: Aventis $30,000

Project Title: Activity of urinary antibiotics against resistant urinary pathogens

PI: George Zhanel, Daryl Hoban, Don Low

Study description: As above

Duration: 2002

Funding: Leo labs $20,000

Project Title: Acute major organ dysfunction in septic patients

PI: Bruce Light

Study description: As above

Duration: 2003

Funding: Glaxo; $85,000

Project Title: Animal models of TSEs

PI: S. Booth

Collaborators: S.Czub, M. Coulthart, G. Telling, K. Qingzhong

Project Description: This project is to establish the infrastructure and expertise within Canada to perform transmission and characterization studies of TSEs in rodent models. Animal models are presently the most sensitive way to establish the presence of TSE infectivity and this capability is essential for the accurate diagnosis and surveillance of TSEs within Canada to inform policy and to assess the risk of transmission between animals and humans. A number of different mouse models, including BSE and CJD that require level III containment, are being established in our laboratory. The study will include primary transmission of Canada’s BSE, CWD and vCJD cases into mice. Transgenic mice containing human, bovine, and elk genes have been previously constructed and reported to show increased sensitivity and short incubation times for transmission. Some of these transgenic models will be used in our studies either in our laboratory or by collaborations with laboratories in the US.

Duration: 2003 onwards

Funding: Health Canada

Project Title: Antibodies to Burkholderia Type III Secretion System

PI: Cangene Corporation (W. Johnson department member)

Co-investigators: Defence R&D Canada; Los Alamos National Laboratory – Dr. Paul Jackson

Project description: B. mallei and B. pseudomallei , two closely related Gram-negative bacteria, are serious potential bioterrorist agents listed on CDC/MMWR’s category B list. These organisms cause life-threatening infections (Glanders’ disease and Melioidosis, respectively), where antibiotic treatment is sometimes insufficient. The virulence factors of the two species are not well understood, and it is difficult to distinguish these species from each other and also from the closely related avirulent species B. thailandensis. B.(pseudo)mallei is also a civilian problem causing septicemia in Southeast Asia. Another Burkholderia species, B. cepacia, is a more common civilian problem that increases mortality and morbidity of cystic fibrosis patients. To better treat and diagnose B.(pseudo)mallei, we will develop monoclonal antibodies (mAbs) against potential virulence factors: components of the type III secretion system (TTSS). The presence of TTSS has been correlated to virulence of Burkholderia. Also, since several other invasive Gram-negative bacteria utilize TTSS to enter host cells, it is plausible that TTSS are critical for Burkholderia’s virulence as well. TTSS components will be cloned and expressed in E.coli to generate sufficient quantities of purified antigen to generate mAbs. Fully human mAbs will be generated by phage display technology. Initial screening of the mAbs for reactivity will be conducted against a panel of Burkholderia isolates cultivated under different conditions. mAbs of interest will be further evaluated for protective capability in both in vitro and subsequently in vivo small animal models. We will also investigate potential differences of TTSS genes among isolates of Burkholderia. In summary, the primary objectives of the project are: 1) Development of mAbs with utility in diagnosing and treating B.(pseudo)mallei and other Burkholderia species, including B.cepacia; 2)Improved understanding of the pathogenicity of B.(pseudo)mallei as a basis for potential vaccine development.

Duration: September 2003 to February 2006

Funding: NIH Grant # 1U10A156383-01; $1,372,000 USD

Project Title: Assay Development and Production team (ADAPT) for the development, validation, production, and distribution of assays for the identification of BT agents.

Co-PI: Jody Berry

Project Description:

Duration: Ongoing

Funding: CRTI-DRDC Project; $3,790,000 over 3 years.

Project Title: Assessing the in vitro Herpes simplex virus inhibitory activity of stannous iron.

PI: Fred Y. Aoki

Collaborators: Dr. Nadia Messiha

Project Description: This in vitro project evaluated the effect of stannous iron on the cytopathic effect of Herpes simplex virus. Stannous iron is the purported antiviral component in a topical cream developed by the sponsor for treatment of recurrent cold sore.

Duration: 2000-2002

Funding: Embro Research; $8,000

Project Title: Averting multiple organ failure in critically illpatients with sepsis: a pilot study of the effects of glutamine supplementation

PIs: Bruce Light, Dr Duerkson, Dr Parry

Study description: As above

Duration: 2001-2002

Funding: STB Research Foundation; $30,000

Project Title: Bacterial Genomics

PI: Michael Mulvey

Study description: As above

Duration: Ongoing

Funding: Canadian Biotechnology Strategy, Health Canada; $650,000

Project Title: Biofilm Build-up in Lumened devices

PI: Michelle Alfa

Duration: 2001-2003

Funding: Steris Inc $80,000 US

Project Title: Bioinformatics capacity

PI : Dr. Lai-King Ng

Project description: To enhance the bioinformatics capacity in the National Laboratory for Sexually Transmitted Diseases.

Duration: Apr 2001 – Mar 2002

Funding: Office of the Chief Scientist, Health Canada; $15,000

Project Title: Biology and epidemiology of hepatitis C virus

PI: J Embree;

co-PI: KM Coombs, F Plummer

Project description: Hepatitis C virus (HCV) is now recognized as a major disease affecting the blood supply and afflicting 170,000,000 people World-wide. Unfortunately, little is known about basic biology of the virus, how it interacts with cells, and risk factors in its transmission from one infected patient to another person. The goals of this study were to determine factors that influence transmission of HCV in a well-characterized patient cohort. Initially, the prevalence of hepatitis C antibodies in mothers and children as well as in groups of prostitutes, were determined, both with and without underlying HIV infection in order to better understand the factors that contribute to passage of the virus from one person to the next. The strains of virus present in the population were also examined to determine if different strains are present in different populations.

Duration: July 2001-June 2002

Funding: Children’s Hospital Research Foundation; $ 22,257

Project Title: Biomedical Proteomics Program: Approaches to the analysis of Disease Progression and Pathogenesis

PI: JA Wilkins;

co-PI: KM Coombs, W Ens, R Beavis, P Nickerson

Project description: Genomics and Proteomics are widely seen as providing powerful analytic methods for determination of alterations in gene expression/utilization. We recently established a Manitoba Centre for Proteomics and have begun to use these mass spectrometric methods for “high-resolution” dissection of virus structure and alterations that take place during assembly and disassembly. We have used this approach to examine details of virus disassembly (Mendez et al., 2003) and to delineate the structure of the RdRp co-factor protein mu2 (Swanson et al, 2002). All other virus proteins are currently under similar analyses. We also are extending these methods to probe protein alterations that take place in cells after and during virus infection. While recent studies are beginning to address genomic alterations (changes in gene expression levels, as measured on “microarrays”), there currently is very little known about what happens to cellular proteins in response to stress. A major new research initiative is to determine changes that take place in viral and cellular proteins during virus infection, both with mammalian reovirus (our model virus) as well as human pathogens like West Nile virus.

Duration: October 2001-September 2004

Funding: CIHR; $ 2,025,594

Project Title: Biosynthesis, targeting, and function of hantavirus glycoproteins

PI: Heinz Feldman

Collaboator: M. Drebot

Project Description: Hantaviruses cause two different clinical syndromes; hemorrhagic fever with renal syndrome (HFRS) which is associated with ‘Old World’ (OW) hantaviruses and hantavirus pulmonary syndrome (HPS) which is associated with ‘New World’ (NW) hantaviruses. Hantaviruses are rodent-borne bunyaviruses that are transmitted to humans from chronically infected mice. The worldwide appearance of HFRS and HPS reflects the geographic distribution of the rodent reservoirs of the different hantaviruses. Sin Nombre (SN) virus, the prototype ‘NW‘ hantavirus, is endemic in Canadian deer mice. HPS cases, although rare in Canada, are of considerable public health concern, because of the high mortality rates associated with SN virus infections. Neither a vaccine nor an antiviral treatment is available. Thus, the identification of determinants for pathogenicity would greatly improve our chances to develop urgently needed therapeutic interventions. Transmembrane glycoproteins are important determinants for the pathogenicity of enveloped viruses. In the case of hantaviruses, the medium segment of the tripartite, negative stranded RNA genome encodes the glycoproteins. Studies on hantavirus glycoproteins were hampered in the past by difficulties with virus propagation and the expression of glycoproteins in mammalian cells. To date, knowledge is mainly derived from studies on the glycoproteins of Hantaan (HTN) virus, the prototype OW hantavirus. Preliminary data obtained from infections with Black Creek Canal (BCC) virus, a ‘NW’ hantavirus, has indicated that intracellular targeting and maturation of the glycoproteins may vary between the two groups of hantaviruses. Based on this data, NW hantaviruses may bud from the plasma membrane in contrast to all other Bunyaviridae which bud from Golgi membranes. However, nothing is known about the biosynthesis and structure of NW hantavirus glycoproteins which are important site determinants for the budding of enveloped viruses. Although it seems clear that hantavirus glycoproteins function in receptor binding and fusion, none of the important functional domains have yet been identified for any of the hantaviruses. In this project, the biosynthesis, structure and maturation of the glycoproteins of NW hantaviruses are studied using eukaryotic expression systems. This will allow us to determine if different maturation processes exist between NW and OW hantaviruses. Reverse genetic systems will are used to identify functional domains on the glycoproteins and to define determinants of pathogenicity. In addition, chimeric proteins are generated and used to define the mechanisms of Golgi targeting and retention.

Duration: 2002-2004

Funding: CIHR; CIHR Regional partnership program; $149,200

Project Title: The burden of lower extremity complications in persons with diabetes and pre or receiving hemodialysis.

PI: John Embil

Project Description: As above

Duration: 2003-2004

Funding: $6,000

Project Title: Canadian Azole Resistance Study (CARS)

Co-PI: George Zhanel and Daryl Hoban

Project Description: Eighteen hospital sites representing tertiary care hospitals in Canada are recruited to provide fungemic Candida spp. isolates from blood cultures (1 per patient) plus Candida spp. isolates from the lower respiratory tract and skin and soft tissue infections. Isolates sent to the reference lab (HSC, Microbiology) for antifungal susceptibility testing. Data analysis to include all susceptibility data plus species prevalence data.

Duration: Ongoing

Funding: Pharmaceutical Industry; $50,000/year

Project Title: Canadian National Fungal Surveillance

Co-PI: George Zhanel, Daryl Hoban

Study description: Surveillance study assessing antifungal resistance in Canada)

Duration: 2001

Funding: Pfizer $120,000

Project Title: Canadian National Fungal Surveillance

Co-PI: George Zhanel, Daryl Hoban

Study description: Surveillance study assessing antifungal resistance in Canada)

Duration: 2002

Funding: Pfizer $120,000

Project Title: Canadian National Fungal Surveillance

Co-PI: George Zhanel, Daryl Hoban

Study description: Surveillance study assessing antifungal resistance in Canada)

Duration: 2003

Funding: Pfizer $120,000

Project Title: Canadian National Fungal Surveillance

Collaborators: George Zhanel, Daryl Hoban

Study description: A surveillance study assessing antifungal resistance in Canada

Duration: 2004

Funding: Pfizer; $120,000

Project Title: Canadian Network for Public Health Intelligence

Co-PI: Amin Kabani

Project description: as above

Duration: 2002 onwards

Funding: CRTI $7,908,234

Project Title: Canadian Network for vaccines and immunotherapeutics of cancer and chronic virus diseases

PI: R. Sekaly; U of Manitoba Collaborators: Keith Fowke, Frank Plummer, Jody Berry

Study description: The goal of this project is to test HIV vaccines

Duration: July 2000 – June 2005

Funding: CANVAC $623,000 .

Study Project: Canadian Nosocomial Surveillance Program Studies

PI: Various: Co-Investigators J Embil, J Embree, M Mulvey et al

Project descriptions:

Nosocomial Cerebral spinal Fluid Shunt-Associated Infections within Acute-Care Institutions: Prospective study of nosocomial CSF shunt infections in participating hospitals across Canada of shunts placed between January 15th 2000 to January 14th, 2002 to determine the incidence, risk factors and outcomes

Vancomycin Resistant Enterococci Suveillance in CHEC/CNISP Health Care Facilities: An ongoing surveillance since 1998 of the incidence and changing trends of vancomycin resistance among Enterococci in Canadian sentininel hospitals. MRSA Suveillance in CHEC/CNISP Health Care Facilities:

An ongoing surveillance since 1995 of the incidence, changing trends and associated risk factors of MRSA in Canadian sentininel hospitals.

Point Prevalence Surveillance for Nosocomial Infections within Selected Canadian Health Care Institutions – February 2002: The variability and cost of nosocomial infections and their prevalence was conducted during a one week period in February 2002 in the 24 CHEC hospitals

Duration: Ongoing

Funding: Health Canada

Project Title: Canadian Paediatric Surveillance Program – Estimating the Incidence of Severe Combined Iimmunodeficiency (SCID) in Canada (2004-March 2006)

PI: Ramsingh R,

Project Team: Pelletier L, Probert A, Yacoub W, Junker A, Schultz K, Champagne MA, Long R, Langley J, Embree J

Project description: A prospective two year study to estimate the incidence of SCID in Canadian children and, in particular, Aboriginal children in Canada to determine the basic demographics, clinical features and outcomes of children diagnosed with SCID in Canada.

Duration: April 2004-March 2006

Funding: Health Canada

Project Title: Canadian Paediatric Surveillance Program – Neonatal herpes simplex virus infection

PI: T Wong;

Co-Investigators: S Burton, J Embree, R Kropp, M Steben

Project description: A study to determine the national incidence of neonatal HSV-1 and HSV-2 infection among Canadian children for the years 2000-2003 and to determine the proportion of those infants with disseminated versus localized disease, the prenatal maternal risk factors among infected infants and to determine the morbidity and mortality attributable to this condition.

Duration: October 2000-September 2003.

Funding: Health Canada

Project Title: Canadian Respiratory Organism Susceptibility Study (CROSS)

Co-PI: Daryl Hoban and George Zhanel

Project Description: Twenty-five labs in Canada are recruited to collect, identify and ship to the reference centre (HSC, Microbiology) S. pneumoniae, H. influenzae, M. catarrhalis, S. pyogenes plus all bacteremic S. pneumoniae. At the reference site microbroth MIC testing is performed using carbapenems, ß-lactams, fluoroquinolones plus other comparators, and data analysis conducted.