Sekundärmetaboliter - Terapeutiska och industriella tillämpningar
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Second messenger molecules - Therapeutic and insutrisal applications (English)
The ability of an organism to adapt to changes in its environment is an essential skill for survival in all kingdoms of life. This led to a vast diversity of pathways that employ intracellular second messengers molecules, whish transduce external signals by modulating intracellular processes. Here, we focus on possible modulations of second messenger signalling pathways for therapeutic and industrial applications.
Project 1: Recent insights into the immune stimulatory effect of cyclic dinucleotides in mammals has widened the scope of their potential therapeutic applications within vaccination, cancer treatment, and autoimmune disease. By fusing the Caulobacter crescentus diguanylate cyclase A (DGCA) to theHIV accessory protein vpr with a pol-derived protease cleavage site linker, we have engineered a HIV-1-derived DGCA protein transducing nanoparticle. Viral protease-mediated release of native DGCA during viral maturation enabled targeted transient delivery of DGCA with the ability to activate the STING pathway via the bacterial second messenger c-di-GMP. DGCA protein transduction demonstrates dose-dependent modulation of STING-mediated activation of URF3 regulatory element containing promoters in transgenic mammalian cells and in immune cells.
Project 2: Heterologous gene regulation systems offer the possibility to increase productivity of biopharmaceuticals and fine-tune production of potentially toxic and difficult-to-produce products. By functionally linking the signal transdcution of the bacterial photo-activated adenylate cyclase bPAC from Beggiatoa sp. PS to the PKA-dependent pathway, and the photo-activateddiguanylate cyclase (DGCL) domain of BphG1 from Rhodobacter sphaeroides to the STING-dependent pathway in mammalian cells, we have engineered a dual wavelength controlled optogenetic expression system for biopharmaceutical production. Activated by blue and near infrared light respectively, transient gene expression from both synthetic pathways can be extensively fine-tuned in transgenic mammalian cells. When scaled for production the dual wavelength controlled expression system displays strong transgene expression with high spatiotemporal resolution when induced by light.
The biological diversity of second messenger signalling pathways provide researchers with the ability to modulate specific cellular functions and construct novel signalling pathways. By employing cross-species and engineered second messenger effector proteins and downstream targets, this work demonstrates two interesting applications for repurposing second messenger molecules and pathways.
Place, publisher, year, edition, pages
second messenger molecules, signalling pathways, cyclic dinucleotides, gene regulation, biopharmaceutical production
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-164188OAI: oai:DiVA.org:kth-164188DiVA: diva2:804978