Diseases caused by dominant, gain-of-function mutations develop in people bearing one mutant and one wild-type copy of the gene. Some of the best known examples of this class are neurodegenerative diseases, including Huntington's, a subset of amyotrophic lateral sclerosis (ALS), Alzheimer's and Parkinson's diseases. In all these diseases, the exact pathways whereby the mutant proteins cause cell degeneration are not entirely clear, but the origin of the cellular toxicity is known to be the mutant protein. Thus, selectively lowering or eliminating the mutant protein is a key step in developing effective therapies. Until recently, it was not clear how specific down-regulation of a wide variety of mutant proteins could be achieved. But now, new advances in RNA interference (RNAi) raise the possibility that RNAi can be developed and eventually applied as a therapeutic means for these neurodegenerative diseases. RNAi can mediate sequence-selective suppression of gene expression in a wide variety of eukaryotes by introducing short RNA duplexes (called small interfering RNAs or siRNAs) with sequence homologies to the target gene. Recent experiments indicate that small hairpin RNAs (shRNAs) transcribed in vivo can trigger degradation of corresponding mRNAs similar to siRNA. These developments raise the possibility that siRNA duplexes or vectors expressing shRNAs may be used to block the expression of a toxic mutant gene. This proposal investigates in vivo efficacy of RNAi therapy using transgenic technology in a mouse model for ALS that is caused by mutations in Cu, Zn superoxide dismutase (SOD1). To determine the potential of RNAi therapy, we will express shRNAs targeting specifically the mutant mRNAs in transgenic mice. We will test how effective and how specific these shRNAs are in suppressing the mutant protein expression and alleviating the disease. To determine in which cell types the suppression of the mutant expression is most crucial, we will express shRNAs in selected cell types using Cre-lox recombination system, We will determine in which cell type suppression of mutant SOD1 expression has the largest impact in alleviating disease. To determine the optimal time for therapy, we will use the Tamoxifeninducible Cre recombinant system to determine at what stage of the disease induction of shRNA to suppress mutant SOD1expresion is most effective.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS048145-04
Application #
7175385
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (01))
Program Officer
Sieber, Beth-Anne
Project Start
2004-02-01
Project End
2009-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
4
Fiscal Year
2007
Total Cost
$348,635
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Biochemistry
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Geng, Chang-Ming; Ding, Hong-Liu (2011) Design of functional small interfering RNAs targeting amyotrophic lateral sclerosis-associated mutant alleles. Chin Med J (Engl) 124:106-10
Yang, Chunxing; Qiu, Linghua; Xu, Zuoshang (2011) Specific gene silencing using RNAi in cell culture. Methods Mol Biol 793:457-77
Qiu, Linghua; Rivera-Perez, Jaime A; Xu, Zuoshang (2011) A non-specific effect associated with conditional transgene expression based on Cre-loxP strategy in mice. PLoS One 6:e18778
Yang, Chunxing; Tan, Weijia; Whittle, Catheryne et al. (2010) The C-terminal TDP-43 fragments have a high aggregation propensity and harm neurons by a dominant-negative mechanism. PLoS One 5:e15878
Wu, Rui; Wang, Hongyan; Xia, Xugang et al. (2009) Nerve injection of viral vectors efficiently transfers transgenes into motor neurons and delivers RNAi therapy against ALS. Antioxid Redox Signal 11:1523-34
Wang, Hongyan; Ghosh, Animesh; Baigude, Huricha et al. (2008) Therapeutic gene silencing delivered by a chemically modified small interfering RNA against mutant SOD1 slows amyotrophic lateral sclerosis progression. J Biol Chem 283:15845-52
Geng, Chang-ming; Ding, Hong-liu (2008) Double-mismatched siRNAs enhance selective gene silencing of a mutant ALS-causing allele. Acta Pharmacol Sin 29:211-6
Qiu, Linghua; Wang, Hongyan; Xia, Xugang et al. (2008) A construct with fluorescent indicators for conditional expression of miRNA. BMC Biotechnol 8:77
Zhou, Hongxia; Falkenburger, Bjorn H; Schulz, Jorg B et al. (2007) Silencing of the Pink1 gene expression by conditional RNAi does not induce dopaminergic neuron death in mice. Int J Biol Sci 3:242-50
Xia, Xu-Gang; Zhou, Hongxia; Xu, Zuoshang (2006) Transgenic RNAi: Accelerating and expanding reverse genetics in mammals. Transgenic Res 15:271-5

Showing the most recent 10 out of 16 publications