TABLE 1.
Drug screens to identify compounds that modulate the actin cytoskeleton and increase ciliogenesis.
| Cell line/origin | Mutational model | Cell culture | Stimulation | Drug library | Methodology | Output (assay markers) | Validation | Hit categories | Specific Hits | References |
| CFPAC-1 | wild-type | 2D | none | Pharmakon 1600 | 10 μM with 2 × 4 day exposure | % ciliated cells (acetylated alpha-tubulin) | assays of hit compounds in four other cancer cell-lines | 118 hits: 49 glucocorticoids, fibrates or other nuclear receptor modulator; 14 neurotransmitter modulators; other include ion channel modulators, tyrosine kinase inhibitors | cilia incidence: clofibrate, gefitinib, sirolimus; cilia length: imexon, clofibrate, xylazine | PMID:26862738 |
| mIMCD3 | Pkd1–/–a | 3D | forskolin treatment | L1200 SelleckChem (273 kinase inhibitors) | 0.1 and 1 μM forskolin for 72 h | cyst size (F-actin and nuclei) | dose response assays of hit compounds | inhibitors targeting mTOR, Aurora A kinase, CDK, IGF-1R, and dual mTOR/PI3K inhibitors | torin 1 and torin 2 | PMID:28644734 |
| MEK | Pkd1–/–, Pkd1+/– and wild-type | 2D screen, 3D validation | none | NIH Pharmaceutical Collection (NPC) and other collections (8814 compounds) | various concentrations over 48 h, followed by comparison between different genotypes | cell proliferation (fluorescence assay), ATP levels (luminescence assay) | re-testing fresh hit compounds, dose response and 3D cyst assays in murine cell; assays in human ADPKD cells and normal kidney cells | 155 hits from 2D screen: HMG-CoA reductase, HSP90, tubulin depolymerization and other inhibitors; 109 hits validated in 3D models; 21 hits validated in human ADPKD cells | 3D models: epothilone A, GSK-269962A, 5-azacytidine, tosedostat; human ADPKD cells: gemcitabine, niclosamide, cerivastatin | PMID:32144367 |
| mIMCD3 | Pkd1–/–b | 3D | none | Cayman Chemicals #10505 (155 kinase inhibitors) | 6 day treatment, various concentrations 1-33 μM | % cord/tubule vs. cysts (F-actin) | treatment of conditional AhCre; Pkd1flox/flox mouse model with Y-27632c | 5 hits: ROCK inhibitors | Y-27632, HA-1077, H-89, (S)-H-1152, (S)-Glycyl-H-1152 | PMID:29891559 |
CFPAC-1, ductal pancreatic adenocarcinoma cell-line; mIMCD3, murine inner medullary collecting duct kidney epithelial cell-line; MEK, murine embryonic kidney cells derived from collecting duct and post-natal proximal tubule cells. aEngineered using lentiviral transfection. bEngineered using CRISPR-Cas9 gene editing. cNo other specific validation of hit compounds; further experiments focused on the RhoA-YAP-cMyc signaling pathway.