Prof. Ilan Tsarfaty

Associate Professor, Department of Clinical Microbiology and Immunology, Sackler Faculty of Medicine, Tel Aviv University

Director, Sackler Cellular and Molecular Imaging Center (SCMIC)

Met Proto-Oncogene and its Ligand, HGF/SF and Breast Cancer

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. 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. We have previously shown that activation of Met by HGF/SF induces an increase in tumor blood volume in a dose-dependent manner. Mimp/Mtch2 reduces cells proliferation in vitro and tumor growth in vivo. Several anti-Met targeted therapies are in development and some have entered phase III clinical trials.

The goal of our studies is to further understand the role of Met-Mimp/Mtch2 in cancer progression and metastasis, and to develop modalities for personalizing targeted Met therapy. 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. 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 is dependent on Met concentration and can be dramatically reduce upon inhibition of the receptor and it’s signaling pathway; Whole animal spectral imaging 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.

Publications

Stein GY, Yosef N, Reichman H, Horev J, Laser-Azogui A, Berens A, Resau J, Ruppin E, Sharan R, Tsarfaty I. Met kinetic signature derived from the response to HGF/SF in a cellular model predicts breast cancer patient survival. PLoS One. 2012. 7:e45969.

Zaritsky A, Natan S, Ben-Jacob E, Tsarfaty I. Emergence of HGF/SF-induced coordinated cellular motility. PLoS One. 2012. 7:e44671.

Shaul, P. Frenkel, M. Goldstein, Mittelman, L., Grunwald, A., Ebenstein, Y. Tsarfaty, I. Fridman, M. The structure of anthracycline derivatives determines their subcellular localization and cytotoxic activity. ACS Medicinal Chemistry Letters 2013. 4, 323-328.

Rivlin, M. Horev, J. Tsarfaty, I. Navon G. Molecular imaging of tumors and metastases using chemical exchange saturation transfer (CEST) MRI. Sci Rep.2013. 25;3:3045.

Zaritsky, A., Manor, N., Wolf, L., Ben-Jacob, E. & Tsarfaty, I. Benchmark for multi-cellular segmentation of bright field microscopy images. BMC Bioinformatics 2013, 14:319.

Laser-Azogui, A., Diamant-Levi, T., Israeli, S., Roytman, Y. & Tsarfaty, I. Met-induced membrane blebbing leads to amoeboid cell motility and invasion. Oncogene 2014. 33:1788-98.

Natan, S*. Tsarfaty, G*. Horev, J. Haklai, R. Kloog, Y. Tsarfaty, I. Interplay between HGF/SF‒Met-Ras signaling, tumor metabolism and blood flow as a potential target for breast cancer therapy. Oncoscience 2014.1:30.

Ninio-Many, L. Grossman, H. Levi, M. Zilber S., Tsarfaty I. Shomron, N. Tuvar, A. Chuderland, M Stemmer, D. S. Ben-Aharon, I. Shalgi, R. MicroRNA miR-125a-3p modulates molecular pathway of motility and migration in prostate cancer cells. Oncoscience 2014. 1:250.

Huang B, Jolly MK, Lu M, Tsarfaty I, Ben-Jacob E, Onuchic JN. Modeling the transitions between collective and solitary migration phenotypes in cancer metastasis. Sci Rep. 2015. 5:17379.

Hecht I, Natan S, Zaritsky A, Levine H, Tsarfaty I, Ben-Jacob E. The motility-proliferation-metabolism interplay during metastatic invasion. Sci Rep. 2015. 4;5:13538.

Hecht I, Bar-El Y, Balmer F, Natan S, Tsarfaty I, Schweitzer F, Ben-Jacob E. Tumor invasion optimization by mesenchymal-amoeboid heterogeneity.Sci Rep. 2015. 5:10622. Erratum in: Sci Rep. 2015;5:12121. 

Zaritsky A, Natan S, Kaplan D, Ben-Jacob E, Tsarfaty I. Live time-lapse dataset of in vitro wound healing experiments. Gigascience. 2015. ;4:8.

Bar-Lev Y, Moshitch-Moshkovitz S, Tsarfaty G, Kaufman D, Horev J, Resau JH, Tsarfaty I. Mimp/ Mtch2, an obesity susceptibility gene, induces alteration of fatty acid metabolism in transgenic mice. PLoS One. 2016;11.

Review

Tsarfaty I, Ben-Jacob E. Secrets of tubule engineering by epithelial cells. Proc Natl Acad Sci USA. 2012. 109:6790-1.