0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
and is designed to determine which of these SNPs are genuinely associated
with ALS. It is envisaged that this study will identify genetic regions and the
actual sequence variations that increase an individual’s risk of developing sporadic
ALS. Thus, the information from this study will guide future ALS
research by identifying novel pathways important in the pathogenesis of
motor neuron degeneration.
Cellular models. The study of cellular models will allow a further screening
and a validation of those genetic factors that may be involved in the
pathogenesis of ALS and allow the individuation of novel pathways modified
in the course of the disease.
Results from this study will hopefully provide suggestions of other candidate
factors, to be translated back to studies in patients.
Animal models. Since the expression of mutant SOD1 is similar in the
in the spinal cord of the two mouse lines we hypothesise that the difference
in the phenotype is due to certain genes that can influence the toxic effect
of SOD1 and can be regarded as genetic risk factors for motor neuron
degeneration. This focused gene expression profiling will provide a framework
for probing specific components of the interactive pathways associated
with the disease. This will add new prospective for effective therapeutic
applications.
OUTPUT(S) OF THE PROJECT
1) Availability of raw genotype data. DNA collection will be complete
within the month 4 of the project. In the following two months, patients and
controls DNA will be processed using the iSelect SNP Chips. The whole
process will take no more than 2 weeks. Within one months, all statistical
analyses will be performed. Raw genotype data generated in this project will
be made publicly available to other researchers, allowing results to be combined
with other data sets by month 6 after the start of the project.
2) Identification of causative mutations. The genes within each candidate
region will be identified and the coding and splice sites of each gene will be
sequenced. We foresee to be able to perform this part of the project within the
second 6 months of the project.
3) Identification of polymorphisms in non coding regions. Immediately
after the previous part of the project, also non coding regions will be assessed.
4) Studies on cellular model of ALS. In parallel with the actions 2 and 3,
motor neuronal cell lines will be manipulated for the silencing of candidate
genes through RNA interference or for the overexpression of candidate genes
through transfection with expression plasmids and used in comparison to the
same cell lines expressing mutant SOD1, to test several parameters of oxidative
stress, mitochondria functionality and protein aggregation that are associated
with ALS. As soon as relevant observation for each gene will be
obtained, these will be made publicly available.
5) Studies on animal models of ALS. In parallel with the actions 2 and 3,
vulnerability of SOD1 mice motor neurons to susceptibility genes identified
740 2009