0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

FISM.9 – Proteomics and metabolomics investigations for the identification of multiple…
with Luca Battistini). To correlate among specific cell-types and proteomics
and/or metabonomics data, we will use evidences collected on a simultaneous
evaluation of eleven different T-cell-associated ‘markers’ (typically cell surface
proteins) in peripheral cells from MS patients in different phases of the disease.
In particular the group of Luca Battistini has identified CD39 as a surface
marker which unequivocally identifies T regulatory cells, and whose
expression confers unique functions. They have also recently shown that
CD39 is expressed by a fraction of CD25bright cells, and that regulatory function
is confined to this subset, while the rest of CD25bright cells are highly
activated conventional T cells, which proliferate vigorously [Borsellino et al.
2007]. CD39 is a member of the family of ecto-nucleoside triphosphate
diphosphohydrolases (E-NTPase family) ectoenzymes, which metabolize ATP
to 5’-AMP, and it is the main enzyme involved in the regulation of extracellular
concentration of nucleotides. Extracellular ATP functions as a danger signal
released by damaged cells as chemo-attractant for lymphocytes, activator
of proinflammatory responses and inducer of local pain. The ectoenzyme
CD39 removes the nucleotide by hydrolytic cleavage and therefore exhibits
immediate immune-modulatory influences on its proximal environment.
Notably, patients with the remitting/relapsing form of multiple sclerosis
(MS) have strikingly reduced numbers of CD39 + Treg cells in the blood. Thus,
in humans CD39 is a marker of a Treg subset likely involved in the control of
the inflammatory autoimmune disease.
In order to investigate on the differentially expressed proteins that potentially
confer suppressive abilities to the CD39+ subset, CD4+CD25bright
peripheral blood cells obtained from healthy individuals will be sorted based
on expression of CD39, and proteins expression from CD4+CD25brightCD39+
and CD4+CD25brightCD39- cells will be analysed by a shotgun proteomics
approach.
The bulk of data obtained with these evidences will be used to build a preliminary
computational model able to predict disease activity in single
patients. To pursue such a goal we defined specific aims which should be
achieved in this proposal:
1) Recruiting in the three clinical sites (Diego Centonze, Alessandra
Lugaresi and Francesco Lolli) of a minimal number of 100 patients divided
between the different MS classes of investigation.
2) Definition of a preliminary univariate and/or multivariate model of
classification of MS patient based on proteomics linear MALDI-TOF mass
spectrometry data from CSF and/or sera.
3) Optimization of cytometric and proteomics data in order to obtain that
combination of biological markers that better identify a diagnostic strategy.
4) Identification of molecular signals from proteomics linear MALDI-
TOF mass spectrometry data from CSF and/or sera enabling patient classification.
5) Definition of a univariate and/or multivariate model of classification of
MS patient based on metabolomics LC-TOF mass spectrometry data from
CSF and/or sera.
2009 625