Annexin A6 and NPC1 regulate LDL-inducible cell migration and distribution of focal adhesions.

Abstract

Cholesterol is considered indispensable for cell motility, but how physiological cholesterol pools enable cells to move forward remains to be clarified. The majority of cells obtain cholesterol from the uptake of Low-Density lipoproteins (LDL) and here we demonstrate that LDL stimulates A431 squamous epithelial carcinoma and Chinese hamster ovary (CHO) cell migration and invasion. LDL also potentiated epidermal growth factor (EGF) -stimulated A431 cell migration as well as A431 invasion in 3-dimensional environments, using organotypic assays. Blocking cholesterol export from late endosomes (LE), using Niemann Pick Type C1 (NPC1) mutant cells, pharmacological NPC1 inhibition or overexpression of the annexin A6 (AnxA6) scaffold protein, compromised LDL-inducible migration and invasion. Nevertheless, NPC1 mutant cells established focal adhesions (FA) that contain activated focal adhesion kinase (pY397FAK, pY861FAK), vinculin and paxillin. Compared to controls, NPC1 mutants display increased FA numbers throughout the cell body, but lack LDL-inducible FA formation at cell edges. Strikingly, AnxA6 depletion in NPC1 mutant cells, which restores late endosomal cholesterol export in these cells, increases their cell motility and association of the cholesterol biosensor D4H with active FAK at cell edges, indicating that AnxA6-regulated transport routes contribute to cholesterol delivery to FA structures, thereby improving NPC1 mutant cell migratory behaviour.

• Authors: Jaimy Jose, Monira Hoque, Johanna Engel, Syed S Beevi, Mohamed Wahba, Mariya Ilieva Georgieva, Kendelle J Murphy, William E Hughes, Blake J Cochran, Albert Lu, Francesc Tebar, Andrew J Hoy, Paul Timpson, Kerry-Anne Rye, Carlos Enrich, Carles Rentero, Thomas Grewal
• Journal: Scientific reports (Sci Rep) Vol. 12 Issue 1 Pg. 596 (01 12 2022) ISSN: 2045-2322 [Electronic] England
• PMID: 35022465 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2022. The Author(s).

Chemical References

• Annexin A6
• Carrier Proteins
• Cholesterol, LDL
• Membrane Proteins
• Niemann-Pick C1 Protein
• STARD3 protein, human
• rab7 GTP-Binding Proteins

Topics

• Animals
• Annexin A6 (metabolism)
• CHO Cells
• Carrier Proteins (metabolism)
• Cell Line, Tumor
• Cell Movement
• Cholesterol, LDL (metabolism)
• Cricetulus
• Focal Adhesions (metabolism)
• Humans
• Membrane Proteins (metabolism)
• Niemann-Pick C1 Protein (metabolism)
• rab7 GTP-Binding Proteins (metabolism)