Compositions and methods for the treatment of diseases associated with aberrant cilia assembly and regulation
View Patent ↗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.