Key words
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Heat-shock proteins; molecular chaperone; protein misfolding; neuro-inflamatory diseases
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Research programs
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COST-Action BM0603 - Inflammation in brain disease
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Short description
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Phase one of the project: We shall use transgenic lines of the land plant P. patens, which we have previously developed to be the most sensitive multicellular eukaryotic organisms to mild heat stresses and to specific chemicals, to perform a simple, amenable, biological screen of about 2000 pharmaceutical compounds. We shall isolate the non-toxic compounds that, within the single micromolar range of concentrations, strongly induce the accumulation of molecular chaperones and proteases. Phase two of the project: We shall test the effect of positive compounds isolated in (1) with a less amenable mobility assay in a C. elegans strain expressing Q35-GFP, which is a temperature-and chaperone-sensitive animal model system for PolyQ-related neurodegenerative diseases (such as Huntington's disease). Phase three of the project: We shall test the effect of positive compounds isolated in (1) and (2) with an even less amenable flying assay in a Drosophila melanogaster assay expressing toxic A?-peptides in neurons, which is a temperature-and chaperone-sensitive animal model system for Alzheimer's disease.
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Further information
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Full name of research-institution/enterprise: Université de Lausanne Département de biologie moléculaire végétale Bâtiment de biologie, room 5427
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Partners and International Organizations
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AT, CH, DE, DK, ES, FR, GR, IL, IE, IT, NL, NO, PL, PT, SE, UK
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Abstract
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During the two-year period, and within the limits set by the finances, we met our first specific aim, which was to isolate non-toxic HSP-inducing drugs in plant cells. Using a reporter line HSP::GUS in the land plant Physcomitrella Patens, we screened a library of about 2000 drugs, pollutants and small organic molecules and isolated about 40 'hit' compounds that could induce mildly or strongly a heat-shock-like response (chaperone accumulation) at isothermal non-heat-shock temperatures. As we were interested only in least-toxic HSP-inducing compounds, we developed a highly sensitive toxicity assay for moss, which was based on a constitutively expressed luciferase that we observed to be highly stable in intact, but not in damaged cells. This toxicity assay reduced our list of hits to about 10 non- or least-toxic HSP-inducing compounds, several of which turned out to be known anti-infalmmatory drugs in mammals. Our second specific aim was to test in an animal system for a known protein-misfolding neurodegenerative desease, some of the 'hits' which were isolated in the moss screen. This aim was partially met: we first used a CL2070 C. elegans strain to test the ability of several 'hit' compounds to induce an isothermal expression of GFP from a heat-inducible sHSP promoter. One compound, a hydroxylamine derivative was further tested in a Caenorhabditis elegans strain modeling toxic (polyQ) protein aggregation leading to muscle paralysis and was shown in motility tests to alleviate paralysis.
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References in databases
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Swiss Database: COST-DB of the State Secretariat for Education and Research Hallwylstrasse 4 CH-3003 Berne, Switzerland Tel. +41 31 322 74 82 Swiss Project-Number: C07.0092
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