© 2007
Freund/OncoLab
The group
Understanding H-prune
biology to fight cancer metastases
Our research follows the
discovery of the H-prune-nm23H1 protein complex and its association to
metastases in cancer. H-prune belongs to the DHH phosphoesterase family
of proteins, and acts as a new phosphodiesterase (PDE) for the cAMP
substrate. Enzyme activity induces cellular motility in a variety of
cancer cells, and likewise, prune overexpression due to chromosome
amplification can lead to metastasis. We are studying this phenomenon at
the cellular level by analysing the motility phases to identify the
mechanism of action in the focal adhesion cellular compartment. We are
looking to identify new protein partners involved in the function of
h-prune. We have found that GSK3-ß, a kinase acting in the WNT canonical
pathway, acts together with h-prune in focal adhesions to promote
cellular motility of colon cancer cells. This complex is influenced
positively by the activity of Rac-GTPases and FA protein kinase. A
second protein interactor, gelsolin, was identified using Tap-Tag
technology coupled to mass spectrometric approaches. This protein is
known to act as a protein severing F-actin in focal adhesions. We are
currently investigating the molecular mechanism of its function with
h-prune in focal adhesions.
We currently have a research focus on the molecular mechanisms of
h-prune action in cellular motility and metastases formation for several
solid tumours, including breast cancer, colorectal cancer, sarcoma,
pancreas carcinoma, hepatocarcinoma, medulloblastoma and neuroblastoma.
For these last two paediatric tumours, a second mechanism of action
together with nm23-H1 and nm23-H2 proteins in the nuclear compartment is
under investigation.
Our laboratory has a technological focus on molecular imaging technology
as a non invasive approach to study tumorigenesis and the prometastatic
role of h-prune in nude mice using orthotopic and xenograft methods.
These experiments are important to define a preclinical animal model for
tumorigenesis with regards to h-prune. We are involved in an
international effort in the fields of structural biochemistry,
crystallography, chemical combinatorial synthesis, pharmacokinetics and
toxicology to identify more selective compounds with structure
similarities to dipyridamole that enzymatically inhibit h-prune. The
preclinical animal models developed in our laboratory are valuable for
preclinical drug testing, and necessary for the translation of research
results into clinical therapies (Phase 1-2).
A PCT international patent protection describing our findings is
registered: PCT (WO/2005/056043): USE OF ENZYMATIC INHIBITORS OF H-PRUNE
FOR THE PREVENTION AND TREATMENT OF THE METASTASES OF TUMOURS
OVEREXPRESSING H-PRUNE (http://www.wipo.int/pctdb/en/wo.jsp?KEY=05/56043.050714).
Functional
NeuroGenomics
Our research
group aims to identify new genes involved in fate specification and
neuronal proliferation and differentiation in the developing
telencephalon. The purpose of the project is subdivided into three main
research lines ranging from a post-genomic approach to the generation
and characterisation of mouse models. We are characterising an unique
collection of genes preferentially expressed in embryonic murine
telencephalon including the discoveries of new microRNAs. Currently, one
focus is directed to identify the human homologues of these genes and
verify a direct correlation to neuro-developmental disorders and cancer
of the brain and peripheral nervous system precusor cells, including
medulloblastoma and neuroblastoma. In particular, we are studying the
roles of newly discovered precursors of microRNAs as pro-differentiation
co-effectors in neuroblasts and cell lines during differentiation in
response to agents, such as retinoic acids and nerve growth factor, or
as pro-apoptotic factors in pro-neuronal cancer-derived cells. We are
validating target gene regulation in vitro using transient expression of
luciferase under the control of target gene promoter sequences, and in
vivo, using orthotopic delivery of human cancer cells into mouse models
for cancer progression (neuroblastoma model). We apply state-of-the-art
gene delivery technology using AntagomiR chemical synthesis in
combination with lentivirus and adeno-associated virus gene therapeutic
approaches in these mouse models.
Our role in EET-Pipeline
Massimo Zollo is Professor of Genetics at the
Faculty of Biotechnological Sciences, Federico II , Naples, and group
leader of the molecular oncology and functional neurogenomics field of
studies at the CEINGE Institution (Naples, Italy). Within the EET-Pipeline,
he is a member of the scientific steering committee and coordinates
efforts to discover new microRNAs involved in paediatric tumours. The
Zollo lab studies the molecular mechanisms involved in cellular motility
and metastasis in cancer, with a particular focus on the functional role
of the human nm23-H1 and -H2 proteins and h-prune in metastasis
formation of neuroblastoma and medulloblastoma. His laboratory is
characterising new dipyridamole analogues for h-prune PDE inhibition in
collaboration with Prof. Aldo Galeone, Faculty of Biotechnological
Sciences (Department of Chemistry, University of Federico II, Naples)
and Prof. Nicola Curtin at AICR (Cancer Research Northern UK Institute,
Newcastle, England.
Forty-eight novel miRNAs specifically expressed during mouse brain
development have been identified by his group to date, and will now be
further analysed in primary embryonal tumours. In cooperation with other
CEINGE researchers, Prof. Zollo provides expertise in different fields
relevant to the E.E.T.-Pipeline project: banking of biological samples,
genome-wide expression analyses, transgenic mouse models, gene-targeting
approaches and cellular and molecular imaging. The presence of the Onco-Genomic
Italian Center sponsored by AIRC at CEINGE headed by Prof. Alfredo Fusco
as well as a close collaboration with the group of Prof. Achille
Iolascon, an expert in clinical paediatric cancer, will further
facilitate the close connection of the E.E.T.-Pipeline to leading
clinicians in the field of study.
Staff Member
People in
Photograph:
On the top,
beginning from the left:
Carotenuto P (Post-doctoral fellow), Aglio V. (Post-doctoral fellow),
Cusanelli E. (PhD Student of European School of Molecular Medicine -
SEMM), Garzia L. (PhD Student of OPEN University), Navas l. (Researcher,
Faculty of Veterinary Clinical Sciences, Federico II, Naples), Prof.
Zollo M. (Group Leader), Prof. Iolascon A. (Group Leader), Orlotti N.I.
(Student of Post-graduated School for Medical Genetics), Cimmino F. (PhD
Student of European School of Molecular Medicine -SEMM).
On the bottom, beginning from the left:
Tagliatatela A. (Under-graduated Student in Biotechnological Sciences),
Maffia V. (Technician), Roma C. (PhD Student of Molecular Oncology,
University of Ferrara, Italy), Scianguetta S. (Student of Post-graduated
School for Medical Genetics), Rossi D. (Under-graduated Student in
Biotechnological Sciences), Marino N. (Post-doctoral fellow).
People involved in EET project
Group Leader
Prof. Massimo Zollo (Associate Professor of Genetics,
Faculty of Biotechnological Sciences, Federico II, Naples, c/o CEINGE,
Centro di Biotecnologie Avanzate,Scarl, Naples, Italy)
Researchers
Luigi Navas (Researcher, Faculty of Veterinary
Clinical Sciences, Federico II, Naples)
Post-docs
Carotenuto Pietro (PhD in Medical Genetics)
Aglio Veruska (PhD in Pharmacology and molecular Oncology)
Marino Natascia (PhD in Pharmacology and molecular Oncology)
PhD Students
Cusanelli E. (PhD Student of European School of
Molecular Medicine - SEMM)
Garzia L. (PhD Student of OPEN University)
Cimmino F. (PhD Student of European School of Molecular Medicine -SEMM)
Roma C. (PhD Student of Molecular Oncology, University of Ferrara,
Italy)
Orlotti N.I. (Student of Post-graduated School for Medical Genetics)
Andolfo I. (Student of Post-graduated School for Medical Genetics)
Bello A.M. (Student of Post-graduated School for Medical Genetics)
Technician
Maffia Veronica
Under-graduated Students
Tagliatatela Angelo (Under-graduated Student in
Biotechnological Sciences)
Rossi
Daniela (Under-graduated Student in Biotechnological Sciences)
Top 5 publications
1. Cancer Cell, 2004. D’Angelo A., Garzia L., Andrč A. , Carotenuto P., Aglio V., Guardiola O., Arrigoni G., Cossu A., Palmieri G., L Aravind and Massimo Zollo. Prune cAMP phosphodiesterase binds nm23-H1 and promotes cancer metastasis.
2. Clinical Cancer Research, 2005. Zollo M. , Andrč A., Cossu A., Sini M.C., D'Angelo A., Marino N., Budroni M., Tanda F., Arrigoni G. and Palmieri G. Overexpression of h-prune in breast cancer is correlated with advanced disease status.
3. Journal of Neuroscience, 2005. Bulfone A, Carotenuto P, Faedo A, Aglio V, Garzia L, Bello A M, Basile A, Andrč A, Cocchia M, Traditi M, Guardiola O, Ballabio A, Rubenstein J and Massimo Zollo. TESS (Telencephalic Embryonic Subtractive Sequences): a Unique Collection of Neurodevelopmental Genes.
4. Molecular Cell Biology, 2006. Kobayashi T, Hino S, Oue N, Asahara T, Zollo M, Yasui W, Kikuchi A. Glycogen synthase kinase 3 and h-prune regulate cell migration by modulating focal adhesions.
5. Oncogene, In press 2007. Garzia L, D’Angelo A, Amoresano A, Knauer S. K, Cirulli C, Campanella C, Stauber R, Steegborn C, Iolascon A and Massimo Zollo.Phosphorylation of nm23-H1 by CKI induces its complex formation with h-prune and promotes cell motility.