Department: Clinical Microbiology and Immunology
Faculty: Medicine-Sackler Faculty
Tel Aviv University

Prof. Tsarfaty Ilan

Research Interest

Nanoparticles based Met-HGF/SF molecular imaging
Breast cancer is the most common malignant disease in western women. In the majority of cases the cause of death in cancer patients is not the primary tumors, but complications derived from metastases at distant sites.  The met proto-oncogene product (Met - a receptor tyrosine kinase) and its ligand, hepatocyte growth factor/scatter factor (HGF/SF), mediate cell motility and proliferation in vitro and tumorigenicity, angiogenesis and metastasis in vivo. Aberrant Met signaling has been widely implicated in most types of human cancers including breast cancer. HGF/SF transgenic mice develop a broad array of histologically distinct tumors. Mutations in the Met tyrosine kinase domain have been identified in both hereditary and sporadic forms of human papillary renal carcinoma.  Mimp/Mtch2, a mitochondrial carrier homologue cloned in our lab, is induced by Met-HGF/SF signaling and is involved in metabolic and bioenergetic processes. Mimp/Mtch2 reduces cells proliferation in vitro and tumor growth in vivo. Over expression of Met and HGF/SF is associated with poor prognosis in several major solid tumor types. Several anti Met targeted therapies are in development and some have entered phase III clinical trials. We have previously shown that activation of Met by HGF/SF induces an increase in tumor blood volume in a dose-dependent manner. 
The goal of our studies is to further understand the role of Met-Mimp/Mtch2 in cancer progression and metastasis, and to develop molecular imaging modalities for personalizing targeted Met therapy. 
Using quantum dots and confocal microscopy we have shown that HGF/SF is bound and internalized by Met expressing DA3 breast adenocarcinoma cells in vitro. Fluorescent tagged–Met proteins were used to study Met mitogenic effect on cells. Met induced cell motility is mediated by the formation of membrane structures such as ruffles, pseudopodia and blebs. Over expression of GFP-Met WT results in its constitutive activation, cell rounding and detachment, and dynamic non-apoptotic membrane blebbing. Bleb retraction results in numerous membrane microspikes where CFP-Met WT, YFP-actin and membrane markers accumulate. Elevation of medium osmolarity by sorbitol inhibits membrane blebbing and increases Met degradation rate. Expression of Dominant-Negative (DN) YFP-Met alone did not induce any membrane blebbing, and co-expression of CFP-Met WT and YFP-Met DN significantly reduces membrane blebbing. Using confocal based molecular imaging we also show that Mimp/Mtch2 reduces the levels of reactive oxygen species ROS and prevents the HGF/SF induced increase in ROS. Mimp/Mtch2 also reduces the polarization of the mitochondrial membrane potential.
To study Met activation by HGF/SF in vivo, we used a xenograft mouse model in which DA3 cells expressing the fluorescent protein mCherry (DA3-mCherry) are injected orthotropicly into mice mammary glands. Contrast media ultrasound-based Met functional molecular imaging (FMI) demonstrated that HGF/SF-induced increased hemodynamics upon over expression and dramatically decreased upon down regulation of the receptor and it’s signaling pathway; Whole animal spectral imaging demonstrated fast developing micrometastatic spread of the tumor enabled detection of sub-millimeter metastases demonstrating fast developing micrometastatic spread of the tumor; Macro to Micro and two photon confocal imaging demonstrated HGF/SF induced changes in blood flow at single vessel resolution, localization of metalloprotease and catapsine activity at the tumor edge and increase in single cell motility.
Met molecular imaging demonstrated that Met signaling modulation plays a major role in breast cancer tumor growth and development. These emerging MI modalities may help tailor Met-targeted therapy.