The overall goal of this project is to discover novel anticancer drugs from endophytic fungi (fungi that live in the intercellular spaces) of desert plants with the broad long term objective of creating a library of this unexploited source of natural products for future drug discovery programs.
The specific aims of the research to be undertaken will include sampling of 30 diverse Sonoran Desert medicinal plants and grasses for endophytic fungi; separating individual fungi and storing these to create a """"""""library of endophytic fungi of Sonoran Desert plants """"""""; culturing 500 endophytic fungi selected from this library on the basis of their diverse taxonomy; processing these fungal cultures and screening the resulting extracts for anticancer activity; fractionating selected bioactive extracts to isolate pure compounds responsible for their observed biological activity; determination of the structure of each biologically active compound; large-scale production of up to 3 compounds with unique structures and novel mechanisms of action followed by evaluation of their in vivo anticancer activity in human tumor xenograft mouse models. The methodologies to be utilized involve the separation and culturing of endophytic fungi in suitable solid or liquid media; successive extraction of fungal cultures with ethyl acetate and n-butanol and screening the resulting extracts in anticancer bioassays. Traditional proliferation/survival assays using three sentinel human cancer cell lines NCI-H460 (lung), and SF-268 (CNS-glioma)} and two new functional assays that monitor the inhibition of angiogenesis and the induction of heat shock protein expression will be used for screening. Dereplication of active extracts will be accomplished by taxonomically identifying the producing fungi based on literature reports and by analyzing bioactive extracts by LC-MS. Bioactivity-guided fractionation of unique active extracts will be performed using solvent-solvent partitioning, gel filtration and chromatography (CC, MPLC, prep. TLC, and HPLC). Structure elucidation of bioactive compounds will be achieved by a combination of spectroscopic (IR, UV, MS, and NMR) techniques, chemical manipulations, or X-ray crystallography as appropriate. Large-scale production (10-20 mg) of up to 3 selected compounds will be undertaken to obtain material for in vivo anticancer activity testing in hormone supplemented SCID mice bearing MCF-7 and T47D human tumor xenografts. We expect that having access to this unique source of bioactive natural products, combined with the use of both conventional and functional in vitro bioassays, followed by in vivo testing will lead to an efficient discovery of new anticancer agents which can be developed into clinically effective drugs with activity against solid tumors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA090265-04
Application #
6855054
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Fu, Yali
Project Start
2002-04-01
Project End
2007-03-31
Budget Start
2005-04-07
Budget End
2006-03-31
Support Year
4
Fiscal Year
2005
Total Cost
$242,169
Indirect Cost
Name
University of Arizona
Department
Miscellaneous
Type
Schools of Earth Sciences/Natur
DUNS #
806345617
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Luo, Jian-Guang; Xu, Ya-Ming; Sandberg, Dustin C et al. (2017) Montagnuphilones A-G, Azaphilones from Montagnulaceae sp. DM0194, a Fungal Endophyte of Submerged Roots of Persicaria amphibia. J Nat Prod 80:76-81
Gubiani, Juliana R; Wijeratne, E M Kithsiri; Shi, Taoda et al. (2017) An epigenetic modifier induces production of (10'S)-verruculide B, an inhibitor of protein tyrosine phosphatases by Phoma sp. nov. LG0217, a fungal endophyte of Parkinsonia microphylla. Bioorg Med Chem 25:1860-1866
Bashyal, Bharat P; Wijeratne, E M Kithsiri; Tillotson, Joseph et al. (2017) Chlorinated Dehydrocurvularins and Alterperylenepoxide A from Alternaria sp. AST0039, a Fungal Endophyte of Astragalus lentiginosus. J Nat Prod 80:427-433
Wijeratne, E M Kithsiri; Gunaherath, G M Kamal B; Chapla, Vanessa M et al. (2016) Oxaspirol B with p97 Inhibitory Activity and Other Oxaspirols from Lecythophora sp. FL1375 and FL1031, Endolichenic Fungi Inhabiting Parmotrema tinctorum and Cladonia evansii. J Nat Prod 79:340-52
Xu, Ya-ming; Bunting, Daniel P; Liu, Manping X et al. (2016) 17?-Hydroxy-18-acetoxywithanolides from Aeroponically Grown Physalis crassifolia and Their Potent and Selective Cytotoxicity for Prostate Cancer Cells. J Nat Prod 79:821-30
Shekhar-Guturja, Tanvi; Gunaherath, G M Kamal B; Wijeratne, E M Kithsiri et al. (2016) Dual action antifungal small molecule modulates multidrug efflux and TOR signaling. Nat Chem Biol 12:867-75
Bai, Jing; Lu, Yuanyuan; Xu, Ya-ming et al. (2016) Diversity-Oriented Combinatorial Biosynthesis of Hybrid Polyketide Scaffolds from Azaphilone and Benzenediol Lactone Biosynthons. Org Lett 18:1262-5
Tillotson, Joseph; Bashyal, Bharat P; Kang, MinJin et al. (2016) Selective inhibition of p97 by chlorinated analogues of dehydrocurvularin. Org Biomol Chem 14:5918-21
Huang, Yu-Ling; Devan, M M Nandi; U'Ren, Jana M et al. (2016) Pervasive Effects of Wildfire on Foliar Endophyte Communities in Montane Forest Trees. Microb Ecol 71:452-68
Tao, Shasha; Tillotson, Joseph; Wijeratne, E M Kithsiri et al. (2015) Withaferin A Analogs That Target the AAA+ Chaperone p97. ACS Chem Biol 10:1916-1924

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