Cancer cell-specific RNA interference in vivo

• Applicant: Professor Dr. Andriy Mokhir
• Subject Area: Pharmacy ; Biological and Biomimetic Chemistry
• Term: from 2019 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 405833972

• Final Report Year: 2023

Final Report Abstract

Technology for suppression of gene expression by small interfering RNAs (siRNAs) led to several clinically approved drugs for metabolic diseases. However, there is still no practical solution for targeting pathological cells within normal tissue by siRNAs. Many disease-associated cells produce elevated levels of reactive oxygen species (ROS), e.g., in cancer and chronic granulomatous disease (CGD), that can be used for their targeting. Though ROS-activated carriers for siRNA delivery are known, they are based on sophisticated, heavily modified polymers and lipids. We developed in this project a simpler prodrug approach based on a minor chemical modification of guide strands of siRNAs with a 5’-O-(4-boronobenzyl)-phosphorothioate group, which is stable during RNA synthesis and under physiological conditions, but is cleaved in the presence of ROS to generate active siRNAs. We demonstrated ROS-responsive activation of the prodrugs for suppression of expression of kinesin family member 11 (KIF11) in vitro as well as initiation factor of protein translation (eEF2) and N-end ruledependent ubiquitin ligases Ubr1, Ubr2, Ubr4, Ubr5 eEF2 genes in vivo.

Publications

• Red light-triggered photoreduction on a nucleic acid template. Chemical Communications, 56(69), 10026-10029.
  Dutta, Subrata; Rühle, Jennifer; Schikora, Margot; Deussner-Helfmann, Nina; Heilemann, Mike; Zatsepin, Timofei; Duchstein, Patrick; Zahn, Dirk; Knör, Günther & Mokhir, Andriy
  
• Translation elongation factor 2 depletion by siRNA in mouse liver leads to mTOR-independent translational upregulation of ribosomal protein genes. Scientific Reports, 10(1).
  Gerashchenko, Maxim V.; Nesterchuk, Mikhail V.; Smekalova, Elena M.; Paulo, Joao A.; Kowalski, Piotr S.; Akulich, Kseniya A.; Bogorad, Roman; Dmitriev, Sergey E.; Gygi, Steven; Zatsepin, Timofei; Anderson, Daniel G.; Gladyshev, Vadim N. & Koteliansky, Victor E.
  
• Intravital electrochemical nanosensor as a tool for the measurement of reactive oxygen/nitrogen species in liver diseases. Journal of Nanobiotechnology, 20(1).
  Abakumova, Tatiana; Vaneev, Alexander; Naumenko, Victor; Shokhina, Arina; Belousov, Vsevolod; Mikaelyan, Arsen; Balysheva, Kamilla; Gorelkin, Peter; Erofeev, Alexander & Zatsepin, Timofei
  
• Reactive oxygen species-responsive RNA interference. Chemical Communications, 58(27), 4388-4391.
  Rühle, Jennifer; Klemt, Insa; Abakumova, Tatiana; Sergeeva, Olga; Vetosheva, Polina; Zatsepin, Timofei & Mokhir, Andriy
  
• Triggering RNA Interference by Photoreduction under Red Light Irradiation. Molecules, 28(10), 4204.
  Rühle, Jennifer; Klemt, Insa & Mokhir, Andriy