Pilot Project Program
Gary K. Ostrander, Ph.D, Program Leader
This Program further enhances research excellence in the area of oceans and
human health. Short-term, modest grant support (of $25,000 to $50,000) is provided
to a maximum of four investigators from varying disciplines each year, to generate
preliminary data in new or emerging areas, which either complement or expand the
currently proposed research projects. In particular, priority is given to pilot
projects, which have a high potential of competing for support from NSF or NIH.
Verbal and written announcements about the Pilot Project Program are distributed
widely through existing community and university networks, encouraging applications
from under-represented minority investigators and women. The evaluation of pilot
project proposals will be based on scientific merit, innovation, qualifications
and academic potential of the applicant investigator, potential contribution of
the research to the general thematic focus and special emphasis areas of the Center,
and overall relevance either locally, regionally or globally.
Pilot Projects
Isolation and Identification of Anti-viral Compounds from Marine Microorganisms
Dr. Yuanan Lu
January 1, 2007 to December 31, 2007
Viruses are small infectious agents which can cause many important human diseases
including SARS, bird flu, hemorrhagic fevers, and AIDS. Viral-induced emerging
and reemerging infectious diseases represent a major health threat to the public.
However, effective control of viral infection and viral diseases has continued
to be a challenge because of the lack of effective vaccine and limited availability
of anti-viral drugs. Hence, new therapeutic measures and antiviral drugs are needed.
One such approach may be to target marine compounds since some marine microorganisms
are known to produce bioactive agents with pharmacological and anti-viral effect,
which could provide unlimited biological resources for the production of therapeutic
drugs for treatment and control of viral diseases in humans, livestocks and marine
farming species. Previous studies from our laboratories have identified several
marine microorganisms with anti-viral potential and also prepared quite a few
of purified bioactive compounds and over 700 crude extracts from a variety of
marine organisms including sponge, bacteria and algae. In this project, we will
set up different cell culture systems and optimize standardized viral assay protocols
for quantitative analysis of several indicator viruses to be included in this
project. Marine compounds and extracts will be pretested in selected cell lines
to determine an effectively safe dose for the use in cell culture. And finally,
we will perform viral neutralization assays to test and screen marine compounds
and extracts prepared from marine organisms for their anti-viral effect. Successful
completion of this study is expected to provide significant data for our understanding
regarding the potential of obtaining anti-viral drugs from marine microorganisms
in our ocean environment.
Marine Resources for Human Health Solutions
Dr. Philip G. Williams
January 1, 2007 to December 31, 2007
One of the most viable Alzheimer's drug targets is BACE1 due to its central
role in the most popular hypothesis of Alzheimer’s disease progression.
As such, pharmacological intervention that partially inhibits BACE1 should have
a valuable therapeutic benefit increasing the quality of life of the patients
and extending the amount of time they remain functionally independent. The discovery
and development of new Alzheimer’s drugs have been slow though. The process
needs to be accelerated and new drug leads need to be discovered now in order
to have effective treatments available within the next 12 years.
Marine cyanobacteria, in particular, are known to be prolific producers of
highly modified peptides and alkaloids including many containing a structural
unit known to inhibit BACE1. We propose to test the hypothesis that a naturally
occurring chemical space consisting of peptide and alkaloids containing these
and other privileged structural fragments from cyanobacteria constitutes a rational
source of new lead compounds for the development of BACE1 inhibitors. To accomplish
this, we will implement an innovative assay protocol that provides an assessment
of the potency of these compounds along with their molecular weight and use this
approach to discover new Alzheimer's drug leads. The advantage of obtaining this
information so early in our approach is that it significantly improves the ability
to prioritize hits based on their likelihood to penetrate the blood-brain barrier,
accelerates the discovery process of natural occurring compounds to one or two
steps, and as a result, drastically shortens the amount of time from hit to structure
identification.
The implications of this research are substantial. It would provide a general
method to accelerate the rate at which novel treatments are discovered from unexploited
natural sources and validate the hypothesis that marine cyanobacteria are a rational
source of protease inhibitors. The benefit is effective Alzheimer’s drug
treatments more quickly.
Marine Microbial Diversity in Hawaii’s Coastal Waters
Dr. Henry G. Trapido-Rosenthal
August 15, 2005 to August 14, 2006
The objective of this Pilot Project is to complement present and future PRCMB
research by using the tools of microbiology, molecular biology, environmental
genomics and educational outreach to attain the following specific aims:
- Study the biodiversity of Hawaii’s coastal marine environments by determining
the taxonomic composition of the microbial populations found therein, using: (a)
the techniques of classical microbiology, coupled to the modern techniques of
molecular taxonomy; and (b) the recently developed techniques of environmental
genomics.
- Determine if the microbial components of the sampled ecosystem contain any
members that might prove to be either deleterious or advantageous to human health.
- Provide sets of microbial cultures and environmental genomic libraries that
can be used as resources in subsequent research programs designed to assess the
relationship of marine ecosystems to human health.
- Develop a cohort of highly trained young Hawaiian biological scientists by
incorporating faculty and students of Kamehameha High School into this project’s
research team, thus enabling them to learn the concepts, techniques, risks and
benefits associated with applying biotechnology to the study of Hawaii’s
biodiversity.
Evaluation of the role of Quorum Sensing Compounds in the Induction of Ciguatoxin
Production by Gambierdiscus toxicus
Paul K. Bienfang, Ph.D.
March 1, 2005 - February 28, 2006
The goal of this program is to identify the variable(s) that trigger production
of ciguatoxin by the G. toxicus via examination for the role of water-soluble
autoinducer compounds in initiating the CTX biosynthesis. This will be pursued
via the execution of laboratory experiments designed to elicit CTX toxicity
in vitro through manipulation of physical-chemical and/or nutrient regimes
at varying cell densities. Identification of a role of such density-dependent,
CTX-inducer compounds will provide new insight that can be applied to further
investigations to improve understanding of the role of environmental triggers
in CTX production. Additionally, the identification of functioning CTX autoinducers
will lead to work to develop improved information bases to mitigate incidences
of ciguatera fish poisoning throughout the tropics. The inclusion of several strategic
partnerships in disciplines associated with CTX chemistry and genomic techniques
allows this program to overcome limitations that have constrained ciguatera experimental
efforts in the past.
Ciguatera fish poisoning (CFP) is caused by the ingestion of fish containing
ciguatoxins (CTX) produced by the epiphytic dinoflagellate, Gambierdiscus
toxicus. Ciguatoxins are biomagnified up through the food web as herbivorous
fish consume the toxic G. toxicus community, and are consumed by carnivorous
fish. Gambierdiscus toxicus, the putative producer of ciguatoxin, settle
onto macroalgae where they grow epiphytically together with a consortium of other
symbiotic microbiota. The ephemeral nature of CTX production both in vivo
and in vitro has been a major obstacle constraining the understanding,
detection and control of ciguatera. Not known are the factor(s) that trigger the
genetically-coupled CTX biosynthesis. Work on the organization of microbial communities
such as biofilms has discovered a sophisticated chemical signaling system for
intercellular communication, called quorum sensing (QS). QS is achieved through
the production, release, and subsequent detection of and response to threshold
concentrations of signal molecules, and may involve either intraspecific or interspecific
communication. Microbial populations frequently require a minimum cell density
for activation of certain functions caused by specific gene expression. Recent
research has shown that expression of many phenotypic characteristics in late-growth
phase cultures, including the production of secondary metabolites and exoenzymes
are cell density-dependent phenomena regulated by QS. The program evaluates the
hypothesis that quorum sensing dynamics are involved in eliciting the gene expression
associated with CTX production by G. toxicus.
Antiinfectives and cytotoxins from Hawaiian marine fungi and yeasts
Stuart Donachie, Ph.D.
August 15, 2004 - August 14, 2006
Marine fungi and yeasts are poorly represented in culture collections. Reliance
upon a few terrestrial fungi and yeasts for most of our products of medical value
risks the development of resistance among pathogens. Intense screening of terrestrial
strains has offered few new leads, so microbes from under-represented taxa and
unexplored sites or niches are required. The overall objective of the proposed
research is to cultivate and screen fungi and yeasts from seawater, marine sediments,
algae and invertebrates in the Hawaiian Archipelago for novel anti-infectives
and cytotoxins.
Specific aims
- To cultivate fungi and yeasts from diverse marine habitats in Hawai‘i.
- To screen cultivated strains for anti-infective and cytotoxic activities.
- To provide lead compound(s) for identification, characterization and development
by collaborators
- To optimize production of promising lead compounds.
Pilot project funds will enable generation of the first data on Hawaiian marine
fungi and yeast diversity, their interactions with endemic invertebrates, and
their bioproducts. These data will support funding requests to the NSF Microbial
Observatories or Microbial Interactions and Processes programs. Neglected
groups such as fungi and yeasts, and especially those in poorly explored habitats,
qualify for the NSF Biodiversity Surveys and Inventories program.
Do microbial biofilm in Hawaiian bays and harbors sequester pathogenic bacteria?
Michael Hadfield, Ph.D.
August 15, 2004 - August 14, 2005
Microbial films are ubiquitous on damp and aquatic surfaces in all environments,
from the most benign to the most extreme. When a clean surface is submerged into
the sea, it is quickly colonized by adsorbent organic molecules and subsequently
with bacteria and other microorganisms. Within a day or two, these biofilms achieve
great complexity in terms of their bacterial compositions and three-dimensional
structures. Marine biofilms are now recognized to be among the most complex in
terms of numbers of bacterial species, although their biodiversity remains greatly
unexplored. We have investigated the biofilms that rapidly develop on new surfaces
in Pearl Harbor, Hawaii and found them to be composed of great numbers of species,
many of which produce critical signal molecules for marine plants and animals
that form a nearly ubiquitous biofouling community across warm-water bays and
harbors world wide.
The overall objective of this study is to locate and identify pathogenic bacterial
species in marine-surface biofilms. For these investigations, biofilms will be
allowed to develop on clean surfaces placed in three or more harbors on Oahu,
for periods of one or more weeks, then analyzed by imaging and molecular techniques,
including confocal microscopy, multiplex PCR, fluorescence in situ hybridization
and denaturing gradient-gel electrophoresis.
|
