A high-throughput screening platform for Polycystic Kidney Disease (PKD) drug repurposing utilizing murine and human ADPKD cells

Abstract

Autosomal dominant polycystic kidney disease (ADpKD) is one of the most common inherited monogenic disorders, characterized by a progressive decline in kidney function due in part to the formation of fluid-filled cysts. While there is one FDA-approved therapy, it is associated with potential adverse effects, and all other clinical interventions are largely supportive. Insights into the cellular pathways underlying ADpKD have revealed striking similarities to cancer. Moreover, several drugs originally developed for cancer have shown to ameliorate cyst formation and disease progression in animal models of ADpKD. these observations prompted us to develop a high-throughput screening platform of cancer drugs in a quest to repurpose them for ADPKD. We screened ~8,000 compounds, including compounds with oncological annotations, as well as FDA-approved drugs, and identified 155 that reduced the viability of Pkd1-null mouse kidney cells with minimal effects on wild-type cells. We found that 109 of these compounds also reduced in vitro cyst growth of Pkd1-null cells cultured in a 3D matrix. Moreover, the result of the cyst assay identified therapeutically relevant compounds, including agents that interfere with tubulin dynamics and reduced cyst growth without affecting cell viability. Because it is known that several ADpKD therapies with promising outcomes in animal models failed to be translated to human disease, our platform also incorporated the evaluation of compounds in a panel of primary ADPKD and normal human kidney (NHK) epithelial cells. Although we observed differences in compound response amongst ADPKD and NHK cell preparation, we identified 18 compounds that preferentially affected the viability of most ADPKD cells with minimal effects on NHK cells. Our study identifies attractive candidates for future efficacy studies in advanced pre-clinical models of ADPKD. Autosomal dominant polycystic kidney disease (ADPKD) has a prevalence of 1 in 500-1,000 individuals and is the most common inherited kidney disease, accounting for 6-9% of patients on renal replacement therapy. ADPKD is caused mainly by mutations in PKD1 and to a lesser extent in PKD2, GANAB and DNAJB11 genes. The disease is characterized by a progressive decline in kidney function due to the formation of fluid-filled cysts as well as activation of inflammatory and proliferative pathways, typically leading to end-stage renal disease by the fifth or sixth decade of life 1. Although cyst formation in the kidneys is the hallmark of ADPKD, other epithelial organs including the liver and pancreas are also commonly affected 2,3. With the 2018 FDA approval of Tolvaptan, a vasopressin V 2-receptor antagonist, there is now one therapy available to slow disease progression; however, the drug was only approved for patients at risk of rapid disease progression due to its potential side effects. Nevertheless, most interventions focus on alleviating disease-related symptoms.

JADE family proteins regulate proteasome abundance and activity

“…In this model, Jade deregulation and the general high proteasomal activity might be one of the mechanisms targeted by Carfilzomib. Proteasome inhibitors have also been very recently identified among other drugs in a study screening about 8000 compounds for their effect on viability of PKD deficient cells (33). Further preclinical studies in mouse models are required to investigate under which exact conditions targeting the proteasome in cystic kidney could be a useful option.…”

Primary Cilia, Ciliogenesis and the Actin Cytoskeleton: A Little Less Resorption, A Little More Actin Please

“…One recent screen assessed the effects of 8,814 compounds, mostly with proven anti-neoplastic effects, on autosomal dominant polycystic kidney disease type 1 (ADPKD1) adult and fetal cell models ( Asawa et al, 2020 ). This screen looked for compounds that reduced cell viability of polycystin 1 ( PKD1 ) null lines but not of wildtype cells.…”

“…The authors then rescreened a selection of compounds for their effects on a 3D cyst culture (unstimulated by cAMP). Although some compounds had no effect on cell viability in 2D, they had significant effects on cyst growth in the 3D model ( Asawa et al, 2020 ). Compounds that affect cell viability or proliferation are unlikely to be effective treatments for all stages of renal cystic disease.…”

An Overview of In Vivo and In Vitro Models for Autosomal Dominant Polycystic Kidney Disease: A Journey from 3D-Cysts to Mini-Pigs

“…As an example, Felekkis et al used, among other cell lines, primary tubular epithelial cells from a transgenic PKD rat model expressing a truncated polycystin-2 protein, to investigate p57/Cdk2 contribution to increased cell proliferation observed in ADPKD cyst lining epithelial cells [ 67 ]. Finally, a very appealing high-throughput approach for ADPKD drug screening has very recently been published by Asawa et al [ 68 ]. In this paper, a comprehensive screening of a large compound library (~8000 compounds) is described.…”