Solo testo.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
analyzed using both a case-control and an intrafamilial association strategies,
as described in our previous work (20,24,32) . In the presence of positive genetic
association, we shall genotype additional SNPs and search for mutations or
chromosomal rearrangements in the SLC25A12 gene. Mutational analysis will
be performed on exons, flanking intronic sequences, 5’ and 3’ UTRs using
DHPLC and DNA sequencing.
Biochemical and gene expression studies – We shall assess asp/glu transport
rates both in gray matter extracts and in mitochondrial pellets, in order to distinguish
functional alterations induced by the cytoplasmic milieu vs changes
intrinsic to ARALAR itself, respectively. Calcium roles will be verified by addition
of EGTA to the medium; SLC25A12 mRNA levels will also be quantified by Real-
Time-PCR, and the extent of correlation between transcription and transport
rates within the same tissues will be assessed. Tissue quality for all assessments
will be verified by total RNA quality assessed using an Agilent Bioanalyser.
The biochemical part of the work program shall be carried out by our collaborator,
Prof. Luigi Palmieri, Ph.D., at the Dept. of Biochemistry of the Univ.
of Bari (Italy), while the genetic part of the work program will be performed at
the Lab. of Molecular Psychiatry & Psychiatric Genetics at IRCCS S. Lucia,
Rome (Italy) under my direct supervision. Prof. Luigi Palmieri is a very wellrenowned
biochemist in the field of mitochondrial transport, with numerous
publications directly contributing to our current knowledge of ARALAR function
(11) and of mitochondrial transporter roles in physiology and disease
(Palmieri et al. (2005) Hum Mol Genet 14: 3079-3088). The current application
is actually being presented to fund only part of the genetic studies involved in
this project, for which additional support is being sought (see additional information
above).
PRELIMINARY RESULTS
Genomic DNA sequencing for the first 3 autistic tissue specimens has been
completed, revealing several polymorphisms, not all listed in available databases
(dbSNP). Biochemical studies performed on the same three pairs have
shown interesting differences between autistic patients and controls, which are
currently being investigated.
POTENTIAL BENEFITS
Patenting benefits – The structure and eventually the functional consequences
of polymorphisms or mutations conferring autism vulnerability will
definitely deserve patenting in view of potential uses as diagnostic tools.
Medical benefits – At present, the disease is recognized exclusively on the
basis of clinical symptoms and, by definition, never before 3 years of age, often
leading to tentative, delayed or even misdiagnosed definitions of patient’s status.
Identification of polymorphisms/mutations in the SLC25A12 gene may
lead to improved and earlier diagnosis through genetic screening, allowing earlier
treatment interventions. In addition, should our studies reveal alterations
696 2006