Kidney Week

Abstract: FR-PO810

Engineering of CX3CR1-Expressing Induced Regulatory T-Like Cells (iTregs) to Treat Acute Glomerulonephritis

Session Information

• Glomerular Diseases: Inflammation and Immunology
  October 25, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

• 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

• Puetz, David L., University of Cologne Center for Molecular Medicine Cologne, Cologne, Nordrhein-Westfalen, Germany

David L. Puetz

• Nies, Jasper Friedrich, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Jasper Friedrich Nies, MD

• Sierra Gonzalez, Claudio, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Claudio Sierra Gonzalez, MS, MSc

• Trinsch, Bastian, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Bastian Trinsch

• Brinkkoetter, Paul T., Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Paul T. Brinkkoetter, MD

• Schermer, Bernhard, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Bernhard Schermer, MD

• Benzing, Thomas, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Thomas Benzing, MD, PhD, FASN

• Diefenhardt, Paul, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Paul Diefenhardt, MD

• Braehler, Sebastian, Universitatsklinikum Koln, Koln, Nordrhein-Westfalen, Germany

Sebastian Braehler, MD

Background

T cell based strategies emerge as a targeted treatment option for autoimmune diseases including glomerulonephritis. T regulatory cells (Tregs) are uniquely equipped to dampen inflammation and thus prevent excessive tissue damage. To harness their full potential, however, they have to be directed to the site of inflammation. The chemokine receptor CX3CR1 detects CX3CL1, which has been shown to be selectively expressed in kidneys during GN. This study aims to engineer CX3CR1 expressing Tregs by retroviral transduction of Foxp3 and Cx3cr1 and assess their suppressive – and ultimately - therapeutic potential in GN.

Methods

A retroviral vector was generated containing the coding sequences of Foxp3 and Cx3cr1 linked by a P2A site. Subsequently, primary naïve murine T cells were transduced in vitro. The generated T cells (termed induced Treg like cells; iTregs) were examined with regards to known Treg markers using flow cytometry. In addition, their suppressive function was assessed in vitro by T cell proliferation assay and ELISA.

Results

Viral transduction of primary mouse T cells resulted in the generation of iTregs expressing FOXP3 as well as CX3CR1 with a high transduction efficiency. Interestingly, these cells expressed the Treg markers CD25, CTLA-4, PD-1, GITR, ICOS and Helios to a similar degree as naturally occurring Tregs. Confirming their regulatory nature, proliferation of T effector cells was suppressed by these iTregs in vitro. In addition, they produced copious amounts of the anti-inflammatory cytokine IL-10, suggesting immunosuppressive functionality.

Conclusion

Retrovirally engineered CX3CR1+ iTregs have suppressive functions in vitro and obtain a Treg like phenotype. Future studies will assess their behaviour and migratory preferences in vivo. Ultimately, their therapeutic potential will be tested in a murine model of acute glomerulonephritis.