Nanoparticle Platform Preferentially Targeting Liver Sinusoidal Endothelial Cells Induces Tolerance in CD4+ T Cell-Mediated Disease Models

Shu-Hung Wang ^1* Isabelle Serr ² Reinaldo Digigow ¹ Barbara Metzler ¹ Alexey Surnov ² Cornelia Gottwick ³ Muhammad Alsamman ¹ Daria Krzikalla ¹ Markus Heine ³ Miriam Zahlten ¹ Agata Widera ¹ Disha Mungalpara ¹ Muharrem Şeleci ¹ Marco Fanzutti ¹ Ligia Margarida Marques Mesquita ¹ Anna-Lisa Vocaturo ¹ Johannes Herkel ^3,4 Antonella Carambia ³ Christian Schröter ¹ Dikran Sarko ¹ Johannes Pohlner ¹ Carolin Daniel ^2,5 Cristina de Min ¹ Sabine Fleischer ^1*

• ¹ Topas Therapeutics GmbH, Hamburg, Germany
• ² Institute of Diabetes Research, Helmholtz Center München, Helmholtz Association of German Research Centres (HZ), Munich, Bavaria, Germany
• ³ University Medical Center Hamburg-Eppendorf, Hamburg, Hamburg, Germany
• ⁴ Hamburg Center for Translational Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Hamburg, Germany
• ⁵ Ludwig Maximilian University of Munich, Munich, Bavaria, Germany

Introduction: Treating autoimmune diseases without nonspecific immunosuppression remains challenging. To prevent or treat these conditions through targeted immunotherapy, we developed a clinical-stage nanoparticle platform that leverages the tolerogenic capacity of liver sinusoidal endothelial cells (LSECs) to restore antigen-specific immune tolerance.Methods:In vivo efficacy was evaluated in various CD4+ T cell-mediated disease models, including preventive and therapeutic models of myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis (EAE), ovalbumin-sensitized delayed-type hypersensitivity (DTH), and the spontaneous type 1 diabetes model. Nanoparticle-induced antigen-specific immune responses were also analyzed through adoptive transfers of 2D2 transgenic T cells into wild-type mice, followed by nanoparticle administration.Results:The peptide-conjugated nanoparticles displayed a uniform size distribution (25–30 nm). Their coupling efficiency for peptides with unfavorable physicochemical properties was significantly enhanced by a proprietary linker technology. Predominant Preferential LSEC targeting of nanoparticles coupled with fluorescently labeled peptides was confirmed via intravital microscopy and flow cytometry. Intravenous nanoparticle administration significantly reduced disease severity and demyelination in EAE, independent of prednisone at maintenance doses, and suppressed target tissue inflammation in the DTH model. Furthermore, prophylactic administration of a mixture of nanoparticles coupled with five autoantigenic peptides significantly lowered the hyperglycemia incidence of the non-obese diabetic mice. Mechanistically, the tolerizing effects were associated with the induction of antigen-specific regulatory T cells and T cell anergy, which counteract proinflammatory T cells in the target tissue. Conclusion:Our findings demonstrate that peptide-loaded nanoparticles selectively preferentially deliver disease-relevant peptides to LSECs, thereby inducing antigen-specific immune tolerance. This versatile clinical-stage nanoparticle platform holds promise for clinical application across multiple autoimmune diseases.