Skip to Content
Merck
  • A human model of Batten disease shows role of CLN3 in phagocytosis at the photoreceptor-RPE interface.

A human model of Batten disease shows role of CLN3 in phagocytosis at the photoreceptor-RPE interface.

Communications biology (2021-02-07)
Cynthia Tang, Jimin Han, Sonal Dalvi, Kannan Manian, Lauren Winschel, Stefanie Volland, Celia A Soto, Chad A Galloway, Whitney Spencer, Michael Roll, Caroline Milliner, Vera L Bonilha, Tyler B Johnson, Lisa Latchney, Jill M Weimer, Erika F Augustine, Jonathan W Mink, Vamsi K Gullapalli, Mina Chung, David S Williams, Ruchira Singh
ABSTRACT

Mutations in CLN3 lead to photoreceptor cell loss in CLN3 disease, a lysosomal storage disorder characterized by childhood-onset vision loss, neurological impairment, and premature death. However, how CLN3 mutations cause photoreceptor cell death is not known. Here, we show that CLN3 is required for phagocytosis of photoreceptor outer segment (POS) by retinal pigment epithelium (RPE) cells, a cellular process essential for photoreceptor survival. Specifically, a proportion of CLN3 in human, mouse, and iPSC-RPE cells localized to RPE microvilli, the site of POS phagocytosis. Furthermore, patient-derived CLN3 disease iPSC-RPE cells showed decreased RPE microvilli density and reduced POS binding and ingestion. Notably, POS phagocytosis defect in CLN3 disease iPSC-RPE cells could be rescued by wild-type CLN3 gene supplementation. Altogether, these results illustrate a novel role of CLN3 in regulating POS phagocytosis and suggest a contribution of primary RPE dysfunction for photoreceptor cell loss in CLN3 disease that can be targeted by gene therapy.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
Lectin from Triticum vulgaris (wheat), agarose conjugate, saline suspension
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Anti-Rhodopsin Antibody, clone 4D2, clone 4D2, from mouse
Sigma-Aldrich
Anti-Retinal Pigment Epithelium 65 Antibody, clone 401.8B11.3D9, Chemicon®, from mouse
Sigma-Aldrich
Anti-Bestrophin Antibody, Chemicon®, from mouse