0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
ratory Standards Institute (CLSI, formerly NCCLS) guidelines [Chace et al.
2007]. We have assessed the pre-analytical factors affecting clinical proteomics
investigations of CSF and sera [Del Boccio et al. 2007]. Moreover
time and cost of analysis for each sample are particularly low, given the ion
source set up, which eventually might handle hundreds of samples in a single
acquisition batch.
2D-PAGE profiling of CSF proteins from patients allows to complement
the dataset provided by linear MALDI-TOF-MS analysis. By this approach
some possible markers of inflammatory activity in MS have been postulated
and, in this line, we have provided preliminary evidence that a tight correlation
between CSF levels of Transthyretin [D’Aguanno et al. 2008] can be found
in MS patients.
Few other studies in mass spectrometry based investigation in Multiple
Sclerosis have been performed [Avasarala et al. 2005; Weingarten et al. 2005].
Unfortunately candidate biomarkers have not been already identified at a
molecular level and the comparison with the CSF protein and metabolites
pattern is still missing.
Metabonomics investigation by mass spectrometry technologies was
already applied in order to investigate characteristic metabolic “ fingerprint
pattern ” as the result of the onset of the disease in different biological fluids
[Wilson et al. 2005; Yin et al. 2006]. Methods development need of a state of
the art high resolution mass spectrometry technology able to obtain high
quality data set and bioinformatics software for data deconvolution and elaboration.
Multivariate statistical analysis (PCA, PLS-DA) allows to highlight differential
metabolic pattern in the metabolite profiles and the information collected
with the described approach could be useful for to better understand
pathological processes.
RATIONALE AND SPECIFIC AIMS
The general picture which emerges from the studies on MS pathogenesis
underlies the notion that all lymphocyte subpopulations are involved in disease
induction and/or progression. It is thus clear that in order to highlight unequivocally
key molecular features of MS progression Proteomics and Metabolomics
data should be analysed in a multidimensional model where the subpopulations
of predominantly involved lymphocytes are taken into account.
The interactions among the various components of the immune system
are indeed very complex, and new technologies have revealed that cellular
populations which apparently seem homogeneous are in fact composed of
several subpopulations with different phenotypes and functions. Such cellular
and functional complexity needs to be properly addressed with novel technologies
able to investigate both the genetic variation and the phenotypic
impact on a large clinical population. This pilot study aim to initiate such a
phenotypic characterisation effort by consolidating the strategic bases for
larger Multicentre Studies and to identify possible markers of MS activity.
In the present research project we will delineate a strategy by which the
different cellular subsets can be unequivocally identified (in collaboration
624 2009