The use of data derived from genomics and transcriptomic to further develop our understanding of Polycystic Kidney Diseases and identify novel drugs for its treatment.
Polycystic kidney disease (PKD) is a prevalent genetic disorder, characterized by the formation of kidney cysts that progressively lead to kidney failure. The currently available drug tolvaptan is... Show morePolycystic kidney disease (PKD) is a prevalent genetic disorder, characterized by the formation of kidney cysts that progressively lead to kidney failure. The currently available drug tolvaptan is not well tolerated by all patients and there remains a strong need for alternative treatments. The signalling rewiring in PKD that drives cyst formation is highly complex and not fully understood. As a consequence, the effects of drugs are sometimes difficult to predict. We previously established a high-throughput microscopy phenotypic screening method for quantitative assessment of renal cyst growth. Here, we applied this 3D cyst growth phenotypic assay and screened 2320 small drug-like molecules, including approved drugs. We identified 81 active molecules that inhibit cyst growth. Multi-parametric phenotypic profiling of the effects on 3D cultured cysts discriminated molecules that showed preferred pharmacological effects above genuine toxicological properties. Celastrol, a triterpenoid from Tripterygium wilfordii, was identified as a potent inhibitor of cyst growth in vitro. In an in vivo iKspCre-Pkd1(lox,lox) mouse model for PKD, celastrol inhibited the growth of renal cysts and maintained kidney function. Show less
Polycystic kidney disease (PKD) is a prevalent genetic disorder, characterized by the formation of kidney cysts that progressively lead to kidney failure. The currently available drug tolvaptan is... Show morePolycystic kidney disease (PKD) is a prevalent genetic disorder, characterized by the formation of kidney cysts that progressively lead to kidney failure. The currently available drug tolvaptan is not well tolerated by all patients and there remains a strong need for alternative treatments. The signaling rewiring in PKD that drives cyst formation is highly complex and not fully understood. As a consequence, the effects of drugs are sometimes difficult to predict. We previously established a high throughput microscopy phenotypic screening method for quantitative assessment of renal cyst growth. Here, we applied this 3D cyst growth phenotypic assay and screened 2320 small drug-like molecules, including approved drugs. We identified 81 active molecules that inhibit cyst growth. Multi-parametric phenotypic profiling of the effects on 3D cultured cysts discriminated molecules that showed preferred pharmacological effects above genuine toxicological properties. Celastrol, a triterpenoid from Tripterygium Wilfordii, was identified as a potent inhibitor of cyst growth in vitro. In an in vivo iKspCre-Pkd1lox,lox mouse model for PKD, celastrol inhibited the growth of renal cysts and maintained kidney function. Show less
Traditional drug discovery approaches have been hampered by (in vitro) cell-culture models that poorly represent the situation in the human body. Principally, cells grow in the body in a three... Show moreTraditional drug discovery approaches have been hampered by (in vitro) cell-culture models that poorly represent the situation in the human body. Principally, cells grow in the body in a three-dimensional (3D) environment that cannot generally be captured using cell culture methods. For this reason, cell-culture models have been developed where cells grow in a 3D-environment, which allows them to form structures that are more comparable to tissue in the body. However, the full complexity of these advanced cell-culture models can only be fully used for routine drug testing if the cell culture model can be used on a large scale (also termed high-throughput screening or HTS), and if the readout can capture all of the biological complexity reflected by the 3D-cultured cells (high-content screening or HCS). Due to these technological limitations, 3D cellular models are not yet routinely applied in drug and drug-target discovery. This thesis describes the development of fully-scalable 3D cell-culture screening platforms in the context of cancer and polycystic kidney disease. Show less
In this thesis we analyzed the pathogenetic sequence of renal cyst and aneurysm formation in PKD mouse models. Cyst formation in the mouse model, Pkd1nl/nl with reduced Pkd1 transcripts was studied... Show moreIn this thesis we analyzed the pathogenetic sequence of renal cyst and aneurysm formation in PKD mouse models. Cyst formation in the mouse model, Pkd1nl/nl with reduced Pkd1 transcripts was studied by analyzing tubular cell proliferation, apoptosis, extracellular matrix remodeling, and the expression the Na+K+2Cl- co-transporter. These mice presented with a rapidly progressive phenotype and, in time, cyst formation was nephron segment-dependent. Interestingly, the expression of Na+K+2Cl- co-transporters was also segment-dependent. TGF__ signaling, the most important cytokine involved in fibrosis, is activated at the progressive stages of the disease and coincides with mild fibrosis and increased expression of TGF__ target genes. These results suggest that the TGF__ signaling pathway is probably not implicated in initial steps of cyst formation, but indicate an important role during cyst progression and in fibrogenesis of progressive ADPKD. Aneurysm formation was found in the Pkd1nl/nl, indicating a direct association between Pkd1 and vessel wall integrity. But, selective disruption of Pkd1 in vascular smooth muscle cells (SM22-Pkd1del/de micel) did not induce any gross structural blood vessel abnormalities. However, SM22-Pkd1del/del mice significantly showed reduced decrease in heart rate upon Angiotensin II-induced hypertension. These findings further demonstrate in vivo, that adaptation to hypertension is altered in SM22-Pkd1del/del mice. Show less
