Cancer targeted treatment by RGD-modified dihydrolypoamide dehydrogenase

Avraham Dayan, School of Molecular Cell Biology and Biotechnology, Tel Aviv University, Tel Aviv, Israel (Avi_idf@hotmail.com)
Gideon Fleminger, School Of Molecular Cell Biology And Biotechnology, Tel Aviv University, Tel Aviv, Israel
Osnat Ashur-Fabian, Department Of Human Molecular Genetics And Biochemistry, Tel Aviv University, Tel Aviv, Israel

The Reactive Oxygen Species (ROS)-dependent phototoxic effect of UV-excited titanium dioxide (TiO2), has been demonstrated in several cancer models of Photodynamic therapy. However, serious damage to the surrounding healthy tissue limits the applicability of this approach. Targeted delivery of TiO2 towards cancer cell would make PDT more selective. Cancer cells often overexpress integrin receptors (e.g. avb3) on their surface, which interact with proteins of the extra cellular matrix through RGD (Arg-Gly-Asp) recognition sites.

Studies in our lab has shown that Dihydrolipoamide dehydrogenase (DLDH) has strong TiO2-binding capabilities. Bio-engineering of DLDH with RGD moieties (DLDHRGD), generated a hybrid-conjugate nanobiocomplex (TiO2-Protein-RGD) capabilities with high affinity to the integrin expressing cancer cells. We have demonstrated that the nanobiocomplex possesses tumor-targeted and UV-excitable cytotoxicity in cutaneous melanoma cells (B16F10) while normal kidney cells (HEK293) remain unharmed.

The activity of mitochondrial dehydrogenases (such as DLDH), is often associated with elevated levels of ROS production, leading to pro-apoptotic activity. Our studies showed that DLDH possesses ROS production activity as well as DNA binding properties. We examined the cytotoxic effects of DLDHRGD and its potential use as an anti-cancer drug with melanoma (B16F10) glioblastoma (005), breast (4T1), cervical (Hela) and ovarian (Ovcar3) cancers cell lines. Normal kidney (HEK293) and cortex (NF5310) were unharmed. DLDHRGD incorporation into the cancer cells and apoptosis induction were analyzed by confocal and FACS assays.

In-vivo assay showed positive safety profile using IV, SC and IP (BALB/C or C57Bl/6 mouse strains). Treatments with DLDHRGD in subcutaneous melanoma mice model resulted in significant tumor inhibition. Currently the efficacy of DLDHRGD on Glioblastoma (murine 005 model in BALB/C) including BBB penetration are under active research.

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Abstract Reference & Short Personal Biography of Presenting Author

Education and degree

• 2019 – Postdoc, Molecular Cell Biology & Biotechnology, Tel-Aviv University.
• 2014-2018 – P.hD, Molecular Microbiology & Biotechnology­, Tel-Aviv University.
• 2012-2013 – M.Sc, Nanomaterials & Nanotechnology­, Tel-Aviv University.
• 2008-2010 – B.Sc, Biotechnology, Bar-Ilan University.
• 2004-2005 – Practical Engineering in Biotechnology and Chemistry, AMAL College.

Publications

• PCT Application (PCT/IL2016/050192) "DLDH, derivatives thereof and formulations comprising same for use in medicine" (2016), National Phase (Aug 2017).
• Avraham Dayan, Gilad Babin, Assaf Ganoth, N. Samir Kayouf, Srijanna Mukkala, Yossi Tsfadia and Gideon Fleminger – The involvement of Coordinative interactions in the binding of Dihydrolipoamide Dehydrogenase to titanium dioxide – localization of a putative binding site. Journal of molecular recognition. (2017).
• Dayan A, Ashur-Fabian O and Fleminger G – RGD-modified Dihydrolipoamide dehydrogenase complexed with titanium dioxide as a novel treatment for cancer. Royal Society of Chemistry – Advances (2018).
• Avraham Dayan, Raphael Lamed, Dafna Benayahu and Gideon Fleminger – RGD-modified dihydrolipoamide dehydrogenase as a molecular bridge for enhancing adhesion of bone forming cells to titanium dioxide implants. Journal of Biomedical Materials Research: Part A (2018).
• Dayan A, Ashur-Fabian O and Fleminger G – Targeted delivery of RGD-modified Dihydrolipoamide dehydrogenase for ROS-dependent anticancer treatment Nature-Oncogene (2019).
• Dayan A, Ashur-Fabian O and Fleminger G – Molecular characterization of dihydrolipoamide dehydrogenase as a DNA binding protein (under minor revision in Langmuir journal)