Progetti

ANTICANCER THERAPY:
Towards a personalized cure using HCS and HCA technologies.

FARNESYLTRANSFERASE INHIBITORS Mode-of-action - AIRC project 1674

Farnesyltransferase inhibitors (FTIs) are emerging anticancer drugs that act on a wide range of malignancies in preclinical tumour models. However, in clinical tests their activity remained restricted to a number of haematological malignancies. As seen today, their wider use will strongly rely on a deeper knowledge of how FTIs affect global cell activity.

Although the target of FTIs is known, the ultimate mechanism by which FTase defect results in FTI anti-proliferative action in Ras-independent tumours remains unclear. The difficulty in assessing FTI mode-of-action is due to the multiplicity of FTase substrates in addition to Ras, among which are RheB, laminins and nuclear associated protein, kinases.

In previous work financed by a CNR-MIUR-Functional Genomic project, we used combined independent approaches of chemical and genetic profiling of yeast cells to assess the pathways that are transcriptionally altered upon FTase inhibition as well as the pathways leading to FTI resistance. By using HCS and HCA technologies we have rapidly validated the results in tumour-relevant cell types.

Future efforts in anticancer therapy will be directed towards:

Developing cell-based assays suitable for High throughput automated cellular image analysis to define the cell cycle and cell proliferation state in tumour-relevant cellular models exposed to clinically relevant anticancer drugs.

Developing diagnostic procedures based on HCA to characterize the cell proliferation state of large numbers of samples.

 

MEMORY and NEURODEGENERATION: molecular basis and drug discovery

Myosin V-based human inherited diseases: Molecular basis of myosin V recognition of intracellular Cargo - Telethon project GGP080143

The study of monogenetic syndromes affecting molecular motors and cargo-bound receptors has allowed deciphering of the basic molecular machinery required for transport within neurons and to clarify the basis of several human multisystemic disorders.  

 Among other cytoskeleton motors acting at neuronal termini, Myosin V (MyoV) was recently shown to be required for the short-range transport of secretory granules carrying neurotransmitters and mRNAs at synapses and nerve terminals. Neuronal plasticity and memory depend on the timely and spatially regulated mobilization of AMPA receptors at dendridic spines of hippocampal neurons. Defects in the trafficking of organelles, mRNA and proteins can result in reduced neuronal plasticity and neuron degeneration.

Defects in MyoVa cause the Griscelli syndrome type 1 (GS1). Griscelli syndromes (GS1, GS2 and GS3) share some common traits: skin and hair hypopigmentation associated with oculocutaneous albinism. In addition GS1 is characterized by severe neurological impairment that is usually of early onset, and includes seizure, mental retardation and hypotonia and ultimately a shortened lifespan (ONIM #214450).

This project aims to clarify how MyoVa mutations lead to neurological dysfunctions typical of GS1. We are creating a cell-based assay in neuronal cell models that will be used in HCS and HCA to identify novel factors able to restore GluR1 trafficking in MyoVa-defective cells. Moreover, by using genetic and biochemical tools applied to yeast cells we are studying the common molecular basis of MyoV-mediated transport in eukaryotes.