Compositions and methods for the treatment of diseases associated with aberrant cilia assembly and regulation
Compositions and methods are provided for identifying agents which have efficacy for the treatment of disorders related to aberrant cilial structure and function, including polycystic kidney disease.
1. A method for identifying agents which modulate ciliary function and assembly/disassembly comprising:
a) providing cells which express AurA, HDAC6 and HEF1;
b) incubating said cells in the presence and absence of a test agent; and
c) assessing activation of HDAC6 and resorption of cilia present on said cells;
wherein an alteration in resorption of cilia or HDAC6 activation in the presence but not the absence of said agent, is indicative of agents which modulate ciliary function and assembly/disassembly.
2. The method of claim 1 wherein the resorption of cilia of step c) is assessed via immunohistochemistry, immunocytochemistry, live cell imaging and/or electron microscopy.
3. The method of claim 1 , wherein said cells are selected from the group consisting of canine kidney polarized epithelial cells (MDCK), mouse kidney collecting duct tubule cells (IMCD3), human renal cells (Caki-1), and human retinal pigmented epithelial cells (HTERT-RPE-1).
4. The method of claim 2 , wherein integrity of cilia is assessed via localization of α-tubulin and γ-tubulin.
5. The method of claim 1 wherein said agent is selected from the group consisting of siRNA, HDAC inhibitors, and small molecule inhibitors of Aurora A kinase.
6. The method of claim 1 wherein said agent promotes ciliary disassembly.
7. The method of claim 1 , wherein said agent inhibits ciliary disassembly.
8. The method of claim 1 , wherein said agent alters a characteristic selected from the group consisting of HDAC6 phosphorylation, HDAC6 deacetylase activity, HDAC6 localization to cilia and length, number and stability of cilia.