Ricerc@Sapienza

From the beginning, Alessio Paone's research activity has been
focused on the study of the function of proteins involved in tumor
processes and their role in cellular metabolic re-programming, with
an approach that ranges from the biochemistry of macromolecules and
their interactors, to molecular and cellular biology. Currently, he is
also directly involved in managing the experimental set up of the
One Carbon Metabolism
Serine
hydroxylmethyltransferase
Cancer cells
HypACB facility at the Department of Biochemical Sciences in
Sapienza, whose service is to gain quantitative respiratory/energetic
parameters of cellular in normoxia and under hypoxic conditions.
More in detail, his research work spna from two main topics:
1) Mechanism of re-shaping of One carbon Metabolism in
cancer.
In the last six years, his work has been focused on proteins involved
in cellular serine/glycine-one-carbon-SGOC metabolism as important
factor in tumor transformation; his study is mainly (but not only)
focused on lung cancer. He is involved in the characterization of the
mechanism behind the SGOC metabolic re-programming, in order to
identify the metabolic status (and vulnerability) of lung cancer cells
required to sustain proliferation. In particular, the role of the enzyme
serine hydroxylmethyltransferase (SHMT), which strongly supports
the production of key molecules for tumor proliferation such as DNA
bases or methyl group donors, has been analyzing by integrating
protein biochemistry and engineering with cell biology.
Main achievement of his research:
- Explaining specific role of the cytosolic SHMT in supporting
the thymine incorporation during DNA replication in lung cancer
cells.
- Characterizing novel role of SHMT as moonlight enzyme. He
contributed in showing that SHMTs are able to interact with high
affinity with RNA; these interactions is required to control both the
expression of the different isoforms of the enzyme and catalysis.
- Identification of novel strategies to target SHMT. Beyond the
physiological relevance of the RNA/SHMT interactions, this feature
allowed the design of novel tool to inhibit the enzymatic activity of
SHMTs. A draft of a patent is currently under evaluation of the
Academic committee.
- Connection between the inflammatory mechanisms regulated
by the TLRs and the metabolic alterations in the tumor cells.
2) "Toll Like Receptors" (TLR) in cancer cells.
During the first post-DOC period, the research focus has been on a
group of proteins called "Toll Like Receptors" (TLR) and their role in
cancer cells. TLRs are receptors of innate immunity and represent the
first line of defense against pathogen invasion. He contributed in
unveiling the mechanism of TLR activation and of the associated
signal transduction process upon interaction of these proteins with
specific ligands in tumor cells.
Main achievement of his research:
- it has been shown that the activation of these receptors can induce
diametrically opposed effects on tumor cells from different tissues
causing in some cases apoptosis, in others proliferation, angiogenesis
etc.
- identification of novel endogenous ligands for TLR7 / 8 with protumor functions. In a work published in PNAS in 2012 a new family
of ligands for hTLR8 and mTLR7 receptors was discovered,
including several mature microRNAs, secreted by tumor cells. These
molecules were able to activate these receptors expressed on the cells 
of the immune system inducing a rewiring in the secretoma which in
turn support the growth of metastases in the lung.
The results reported were also obtained thanks to the skills acquired
by Alessio Paone specifically in the study of:
- protein-protein and protein-RNA interactions;
- signal transduction;
- processes of cell death;
- regulatory metabolic mechanisms.
By taking advantage of his technical expertise in:
-protein biochemistry and engineering: expression and purification of
proteins.
-cell biology: cell culture, Western Blotting, RT-PCR, iRNA,
fluorescence microscopy, respiratory assays through Seahorse
apparatus.
-in vivo: mouse handling, intraperitoneal injections, xenograft tumor
inoculation, in vivo angiogenetic assays.
-data handling: standard statistics, seahorse data analysis