Vinculin influences essential processes in enteric nervous system development and Hirschsprung disease pathogenesis

Area of Science:

• Developmental Biology
• Genetics
• Cell Biology

Background:

• Vinculin (VCL) is a key cell-cell and cell-matrix adhesion protein.
• Its role in disease, particularly enteric nervous system (ENS) disorders like Hirschsprung disease (HSCR), is largely unexplored.

Purpose of the Study:

• To investigate the role of VCL in ENS development.
• To determine the association between VCL mutations and HSCR.

Main Methods:

• Whole-genome sequencing to identify VCL mutations.
• Generation of neural crest-specific Vcl knock-out (Vcl cKO) mice.
• Single-cell and spatial RNA sequencing.
• In vitro assays.

Main Results:

• Four VCL mutations linked to HSCR were identified, mostly causing loss of function.
• Vcl cKO mice exhibited ENS defects mimicking short-segment HSCR.
• Dysregulation of neuronal differentiation and MAPK signaling genes was observed.
• Reduced ENS-mesenchyme interactions and Pleiotrophin (PTN) pathway disruption were found in Vcl cKO mice.

Conclusions:

• VCL is a crucial hub gene for ENS formation, essential for cell adhesion and signaling pathways.
• VCL deficiency leads to ENS defects through subtle impacts on cellular interactions and signaling, resulting in HSCR-like phenotypes.
• The findings highlight the importance of VCL, MAPK, and PTN pathways in ENS development and suggest potential therapeutic targets for ENS disorders.