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  3. Recreating pathophysiology of CLN2 disease and demonstrating reversion by TPP1 gene therapy in hiPSC-derived retinal organoids and retina-on-chip

Recreating pathophysiology of CLN2 disease and demonstrating reversion by TPP1 gene therapy in hiPSC-derived retinal organoids and retina-on-chip

  • Cell Rep Med. 2025 Aug 19;6(8):102244. doi: 10.1016/j.xcrm.2025.102244.
Serena Corti 1,  Kwi Hye Kim 2,  Ting Chen 2,  Adelina Botezatu 1,  Virginia Cora 1,  Ke Ma 1,  Natalia Pashkovskaia 1,  Anamaria Bernal Vergara 1,  Denise Sperlich 1,  Kaushambee Dave 1,  Arianna Tolone 1,  Ryan M Reddinger 2,  Christopher B Tully 2,  Mikayla Higgins 2,  Alexander Kleger 3,  Markus Breunig 4,  Paul Lopatta 4,  Svenja Wingerter 5,  Madalena Cipriano 5,  Sylvia Bolz 6,  Marius Ueffing 6,  Nicholas Buss 2,  Peter Loskill 7,  Stefan Liebau 1,  Kevin Achberger 8
Affiliations

Affiliations

  • 1 Institute of Neuroanatomy & Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany.
  • 2 REGENXBIO Inc, Rockville, MD, USA.
  • 3 Institute of Molecular Oncology and Stem Cell Biology, Ulm University Hospital, Ulm, Germany; Division of Interdisciplinary Pancreatology, Department of Internal Medicine I, Ulm University Hospital, Ulm, Germany; Core Facility Organoids, Ulm University, Ulm, Germany.
  • 4 Institute of Molecular Oncology and Stem Cell Biology, Ulm University Hospital, Ulm, Germany.
  • 5 Institute of Biomedical Engineering, Eberhard Karls University Tübingen, Tübingen, Germany.
  • 6 Centre for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University Tübingen, Tübingen, Germany.
  • 7 Institute of Biomedical Engineering, Eberhard Karls University Tübingen, Tübingen, Germany; NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
  • 8 Institute of Neuroanatomy & Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany. Electronic address: kevin.achberger@uni-tuebingen.de.
PMID: 40706588 DOI: 10.1016/j.xcrm.2025.102244
Abstract

Mutations in the tripeptidyl peptidase 1 (TPP1) gene lead to neuronal ceroid lipofuscinosis type 2 (CLN2), characterized by lysosomal accumulation of lipofuscins predominantly in the brain and retina. The ocular phenotype is characterized by outer retinal degeneration that leads to vision loss. Leveraging human induced pluripotent stem cell (hiPSC)-derived retinal organoids (ROs), retinal pigmented epithelial cells, and the retina-on-chip system, we establish an in vitro CLN2 model that recreates the principal histological hallmarks, namely the accumulation of subunit C of mitochondrial ATP Synthase (SCMAS) and lipids mainly in the outer retina. Furthermore, single-cell RNA Sequencing reveals a dysregulation of translational and mitochondrial function in CLN2 cones. Finally, adeno-associated virus (AAV)-mediated TPP1 gene therapy can restore TPP1 expression and decrease and even prevent SCMAS accumulations. Our study uses an innovative human-relevant microphysiological retinal disease models, uncovers previously uncharacterized mechanisms of CLN2 pathophysiology, and demonstrates the potential of AAV9.hCLN2 gene therapy for CLN2 disease, potentially treating patient blindness.

Keywords

CLN2; SCMAS; TPP1; gene therapy; neuronal ceroid lipofuscinosis type 2; retina-on-chip; retinal organoids; subunit C of mitochondrial ATP synthase; tripeptidyl peptidase 1.

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