Tailored gene therapy to generate liver-protective designer NK cell micropharmacies

• Applicants: Professor Dr. Michael A. Morgan; Professor Dr. Axel Schambach
• Subject Area: Hematology, Oncology ; Cell Biology
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 564806462

Designer cells can be used to overcome challenges that limit the effectiveness of liver organ transplantation, such as inflammation/inflammatory signaling, rejection and fibrosis, and thus improve graft longevity and fitness. Liver fibrosis is a progressive disease characterized by excess extracellular matrix proteins and a main factor that directs the pathologic processes of liver fibrosis is activation of hepatic stellate cells (HSCs) in response to a proinflammatory liver microenvironment. Here, upregulation of fibroblast activation protein (FAP) on activated HSCs drives fibrosis and FAP-positive HSCs serve as promising therapeutic targets. Our goal is to engineer designer NK cells with the ability to home to the liver, serve as local therapeutics and to secrete protective and regenerative tailored therapeutics directly within the liver, e.g. as a living micropharmacy. Designer NK cells will be generated from stem cell/progenitor-derived and primary NK cells. To direct designer NK cells to home to the liver, NK cells will be engineered to express chemokine receptors (e.g., CXCR6, CCR5, CXCR3) and transcription factors (e.g., Hobit, T-bet, Eomes) known to be expressed by liver-resident (lr) NK cells. Designer NK cells that are responsive to local environmental cues, including danger signals, inflammatory signaling and fibrosis will be engineered. In addition, we will express immunomodulatory, regenerative and anti-fibrotic factors to save troubled liver transplants. Our main hypothesis is that precision gene therapy principles can be used to generate transplantable designer NK cells to promote liver organ transplant tolerance and survival by counteracting inflammatory signaling and fibrosis in the area of the graft. Designer NK cells will support organ regeneration by targeted delivery of inducible cytokines, danger signal scavengers, growth factors and healthy tissue-supporting secretable miRNAs to the graft using our established inducible systems based upon vectors we developed for TRUCKs (“T cells redirected for antigen-unrestricted cytokine-initiated killing”) coupled with a chimeric antigen receptor that recognizes FAP-positive cells validated in our laboratory. Inducible and targeted delivery of protective strategies will be followed in 2D and 3D hepatocyte models and transplantation into ex vivo perfused livers and into suitable animal models to explore the targeting and regenerative capacities of our approach. The long-term goal of this project is to generate designer immune cells that can be used to improve liver organ transplantation using regenerative, anti-fibrotic and anti-inflammatory signaling cues to enhance organ health. The principles used and developed in this project can be expanded for application to other organ systems, and can be translationally exploited.

• DFG Programme: Research Grants