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Natural products from marine fungi for the
treatment of cancer Johanna Silber, Johannes F. Imhoff, Antje Labes
Marine fungi as a good source for new natural products
Though marine fungi are a potent group of secondary metabolite
producers, they are not well characterised and underutilised in
terms of biotechnological application. Here, we demonstrate the
sustainable exploitation of marine natural resources providing
appropriate culture conditions for the group of marine fungi, thus
enabling efficient production of marine natural products in the
laboratory and also in large scale cultures, avoiding harm to the
natural environment. In the focus are new anti-cancer compounds.
Two approaches are used to gain effective producer strains (fig. 1):
a) Candidate strains originating from our unique strain collection of
marine fungi are characterised and optimised using molecular
methods. The genomes of these strains are sequenced during the
project (fig. 3).
b) New fungi are isolated from unique habitats, i.e. tropical coral
reefs, endemic macroalgae and sponges from the Mediterranean.
Culture conditions for these new isolates are optimised for the
production of new anti-cancer metabolites.
Fig. 1 MARINE FUNGI is aimed to develop a
process concept providing the technological
basis for a sustainable use of marine microbial
products in oncology.
Characterisation of candidate structures to the
stage of in vivo proof of concept
The development of clinically relevant lead structures
requires affords beyond screening: An interdisciplinary
approach is necessary to characterise hit candidates to the
stage of in vivo proof of concept ready to enter further drug
development in order to valorise the results of the project.
This includes the development of robust, sustainable
processes for the production of these compounds.
Therefore, the Kiel Centre for Marine Natural Products
(KiWiZ) initiated and coordinates an EU FP7 research
project, MARINE FUNGI, focussing on the development of
anti-cancer drugs from marine fungi (fig. 1). MARINE
FUNGI covers two approaches to gain effective producer
strains, which will be led to the stage of in vivo proof of
concept building the basis for clinical trials (fig. 1, 2).
Bildunterschrift in hellblau
Fig. 2 MARINE FUNGI formed of a new strongly interacting research network comprising the scientific and technological
actors, including 3 SMEs and 2 ICPC partners. Corals, algae and sponges are the sources for new fungal metabolites.
More than 500 isolates are screened for their secondary metabolites until the end of 2012.
Current status of MARINE FUNGI
• Three fungal genomes are sequenced. The search for
biosynthetic and regulatory genes is ongoing.
• More than 500 new fungal isolates were gained, taxonomic
affiliation is done by molecular and microscopic methods.
• New strains are cryoconserved and established as the projects
strain collection.
• Nearly 2500 extracts were screened in the preliminary panel.
• 50 compounds were screened in the full panel leading to a
selection of 6 compounds ready for liability testing.
• A process concept for these compounds is build comprising
fermentation in stirred tank reactors and subsequent purification.
• S. brevicaulis is oprimised by mutagenesis experiments, a library
of UV mutants was established.
• 8 PhD students are involved in MARINE FUNGI.
• A website was set up, a weekly blog informs about news.
WP1/2: Project Management and Ccoordination
WP5 Chemical identification and
biochemical characterisation of active metabolites and substance purification
WP3 Genome analysis, identification of biosynthetic
genes and regulators
WP7 In vitro bioassays for cancer targets, rational lead structure selection and in vivo efficacy
determination in xenograft models
WP8 Robust and sustainable process
development
WP9: Intellectual Property protection & dissemination activities
WP6 Strain improvement
Selected fungal strains Selected macrobes from unique marine habitats
Culture based approach Molecular based approach
WP4 Isolation and identification of new fungal strains
Teknologisk
Institut
11 partners, 7 countries
Consortium of
MARINE
FUNGI
For further information visit: www.marinefungi.eu
Calcarisporium sp. Scopulariopsis brevicalis Pestalotiopsis sp.
Gene prediction
Commercial sequencing of fungal genomes
Preprocessing of raw sequence reads
454 GS-FLX Titanium Ion-torrent
Lucy/seqClean
Hybrid genomic assembly
CLCBio/MIRA3
Genome quality assessment
Fungal genomic features
OK
General Annotation
BLAST2GO
Protein Domains
InterPro/Pfam
Enzymatic pathways
KEGG
Secondary Metabolite clusters
SMURF
Extraction of various genetic features
not OK
Illumina Hiseq 2000
AUGUSTUS
N
NNHNH
N
OCH3
CH2
CH3
OH
OO
CH3
Fig. 3 Three fungal strains were chosen for genome sequencing.
The genomes will help to identify biosynthesis genes and give
insights into regulatory features.
Fig. 4 All extracts and pure compounds
originating from the initial screening and
subsequent purification are screened against
various panels of cancer cell lines.
M14 melanoma
MCF7 breast
HL-60 leucemia
786-0 renal
Preliminary panel for extract screening
Dereplication of meleagrin as example for structural
elucidation