Project Details
Projekt
Primary cilia dynamics in retinal pigment epithelium development, homeostasis, and function
Applicant
Professorin Helen May-Simera, Ph.D.
Subject Area
Cell Biology
Developmental Biology
Developmental Biology
Term
since 2023
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 503306912
The retinal pigment epithelium is a prime target to understand the molecular mechanisms of how temporal and spatial regulation of cilia dynamics control epithelial organization. This monolayer of highly specialized, tightly connected polarized cells located between the neural retina and the vascular choroid is essential for vision. Similar to other epithelia, the primary cilium in the RPE is a highly dynamic organelle, that regulates many signaling pathways controlling RPE specification, development, and differentiation. Of these, the WNT signaling pathway seems to be one of the most prominent. To what extent the precise regulation of cilium assembly and disassembly dynamics influence RPE development, maturation, or function via regulation of WNT signaling is unknown. To address these questions, we propose the following three specific objectives. 1. Investigate the molecular mechanisms of RPE maturation and function via cilia-mediated WNT signaling in vitro. 2. Compare cilia ablation vs. altered ciliary signaling in RPE cells in vivo and analyze the consequences for retina and visual function. 3. Characterize cilia assembly and disassembly in real-time explant tissue cultures. Our particular focus is to distinguish the relevance between ciliation vs. ciliary signaling and explore how cilium assembly and disassembly can influence RPE maturation and function. Using well-established RPE cell models (in vitro) and RPE-specific cilia mouse mutants (in vivo) and we will combine gene knockout/silencing and pharmacology to inhibit ciliation and manipulate ciliary signaling. In each of these systems we will manipulate three ciliary proteins (BBS8, IFT20, IFT88) to either modulate ciliary signaling or ablate the cilium. Our three aims are interrelated but not co-dependent. Upon completing the first two aims, we will discover whether the primary cilium assembly/disassembly dynamics are key in controlling RPE differentiation and maturation and which ciliary signaling pathways dynamically regulate this process. By comparing the phenotype of these we can distinguish between ciliation vs. ciliary signaling effects. In the third aim we will establish a platform for further downstream analyses. Insights into how primary cilia dynamics regulate RPE organization is not only crucial for vision research but can also be applied to other epithelial tissues.
DFG Programme
Research Units
