A University of Pittsburgh researcher has developed a novel approach to increase mRNA-mediated protein expression following nanoparticle liposome-mRNA (NPL-mRNA) delivery. A modified form of human gasdermin C (GSDMC) can aid the release of mRNA from liposomes. By delivering the mRNA for the modified GSDMC alongside the therapeutic mRNA in the same liposome, it is possible to enhance the release of therapeutic mRNA from the nanoparticle liposome and the subsequent expression of the desired protein. This novel approach could revolutionize the clinical uses and efficacy of mRNA-based vaccines or therapy.
A modified active form of GSDMC (Flag-289) can facilitate mRNA-mediated protein expression. Delivery of the mRNA for Flag-289 along with therapeutic mRNA in a liposome, results in expression of the “helper” Flag-289 alongside the therapeutic mRNA expression. Flag-289 can lyase more mRNA-containing endosomes, enhancing the efficacy of NLP-mRNA therapy.
Description
NPL-mRNA vaccines played a crucial role in managing the COVID-19 pandemic. Since then, mRNA-based vaccines have been increasingly explored as vaccines against other conditions such as tuberculosis and some cancers. NPL-mRNA also has the potential to treat a host of conditions where key proteins are not produced endogenously (e.g., factor VIII in hemophilia A). Following transfection with NPL-mRNA, the mRNA must escape to undergo transcription and produce the desired protein. Modified GSDMC can facilitate the release of mRNA improving the efficacy of any NPL-mRNA treatment; this discovery may lead to the development of more mRNA-based therapies to include targeting cell lines traditionally difficult to transfect.
Applications
• Vaccinations
• Protein deficiency syndromes
Advantages
Rab7 is a small G-protein that regulates late endosome transport. Recently it was discovered that a cleaved form of GSDMC N-terminus can penetrate Rab7+ late endosomes and lyase the endosome, releasing the encapsulated content (e.g., mRNA). However, this active form of GSDMC can damage plasma membranes causing cell death in a process known as pyroptosis.
A modified version, N-flag-tagged GSDMC N-terminus (Flag-GSDMCN-ter) will lyase endosomes but not induce pyroptosis, to facilitate mRNA-mediated protein expression without causing cell death. This novel treatment strategy delivers both the “helper” mRNA encoding for Flag-GSDMCN-ter and therapeutic mRNA encapsulated in NPLs. Upon cell entry the “helper” mRNA is expressed, boosting mRNA transfection efficiency to increase therapeutic benefit.
Invention Readiness
A Flag-GSDMCN-ter (Flag-289) was generated using a protease, Cathepsin S. In vitro studies confirmed Flag-289 could penetrate Rab7+ vesicles. Penetration by RAB7+ compromises the vesicles easing release of the mRNA cargo. Further experiments confirmed Flag-289 could boost transfection efficiency in NIH3T3, LLC, and MC38 cell lines.
IP Status
Patent Pending
