Biology subjects

Sander, V.

Publications and source records attributed to Sander, V..

5 recordsLinked to original sources

bioRxiv2025

High efficiency organoid-derived cyst cultures as a drug discovery platform for polycystic kidney disease

Autosomal dominant polycystic kidney disease (ADPKD) causes the slow, progressive formation of fluid-filled cysts within the kidney that ultimately leads to renal failure. Despite being the most common genetic kidney disease, the range of therapies for ADPKD is limited. Ureteric bud (UB) organoids derived from human pluripotent stem cells (hPSCs) have been used to model ADPKD. Here, we describe a protocol to generate UB organoids in large-scale suspension culture that is amenable to drug discovery. When differentiated from PKD2-deficient hPSCs, the UB organoids robustly form cysts within 7 days and reach nearly 100% cyst efficiency within [~]20 days. Cystic UB organoids express ADPKD markers and are responsive to cyst-reducing compounds, such as the BET inhibitor JQ1 and the NF-{kappa}B inhibitor QNZ, but not tolvaptan. As proof-of-principle of the platform as a drug discovery tool, we screened a small molecule library comprised of nutrient conjugates and identified M1, a palmitate-glucosamine-glycine conjugate, that reduced cyst formation. These results demonstrate that our scalable method of cystic UB organoid production provides a simple and highly efficient platform for ADPKD drug discovery. TRANSLATIONAL STATEMENTAutosomal dominant polycystic kidney disease (ADPKD) affects over 12 million people worldwide, yet the currently available therapy tolvaptan only moderately delays disease progression and is burdened with adverse effects. Improved treatments that are well tolerated and effective are urgently needed. We established a human ureteric bud organoid platform that enables ADPKD-like cysts to be cultured at large scale and low cost. These cystic organoids resemble human ADPKD cysts and are suitable for drug discovery. Identification of the nutrient conjugate M1 as a new lead molecule for reducing cyst formation highlights the clinical relevance of this model.

molecular biology↗

bioRxiv2024

Partial repair causes permanent defects in papillary structure and function after reversal of urinary obstruction

Urinary obstruction causes injury to the renal papilla and leads to defects in the ability to concentrate urine which predisposes to progressive kidney injury. However, the regenerative capacity of the papilla after reversal of obstruction is poorly understood. To address this, we developed a mouse model of reversible urinary obstruction which is characterized by extensive papillary injury, followed by a robust regeneration response and complete histological recovery over a 3- month period. However, these mice have a pronounced defect in urinary concentrating capacity. We now show that this is due to permanent changes in the composition, organization, and transcriptional signatures of epithelial, endothelial, and interstitial cell lineages in the papilla. There are persistent inflammatory responses that are also seen in patients with renal stone disease but are associated with cell-specific adaptive responses to the increasingly hypoxic environment of the papilla after reversal of obstruction. Taken together, our analysis of a new model of reversible urinary obstruction reveals that partial repair leads to permanent changes in the structure and function of all of the major cellular compartments in the papilla that include both shared and distinct responses to different types of renal papillary injury in humans and mice. SummaryPartial repair after reversal of urinary obstruction leads to permanent changes in structure and function of all major cellular compartments in the renal papilla O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/612436v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@1438af5org.highwire.dtl.DTLVardef@157ff60org.highwire.dtl.DTLVardef@3f09c0org.highwire.dtl.DTLVardef@9950b1_HPS_FORMAT_FIGEXP M_FIG C_FIG

pathology↗

bioRxiv2021

Nephrotoxicity of the BRAF-kinase inhibitor Vemurafenib isdriven by off-target Ferrochelatase inhibition

A multitude of disease and therapy related factors drive the frequent development of renal disorders in cancer patients. Along with chemotherapy, the newer targeted therapeutics can also cause renal dysfunction through on and off-target mechanisms. Interestingly, among the small-molecule inhibitors approved for the treatment of cancers that harbor BRAF-kinase activating mutations, vemurafenib can trigger tubular damage and acute kidney injury (AKI). To investigate the underlying mechanisms, here, we have developed cell culture and mouse models of vemurafenib nephrotoxicity. Our studies show that at clinically relevant concentrations vemurafenib induces cell-death in transformed and primary murine and human renal tubular epithelial cells (RTEC). In mice, two weeks of daily vemurafenib treatment causes moderate AKI with histopathological characteristics of RTEC injury. Importantly, RTEC-specific BRAF gene deletion did not influence renal function under normal conditions or alter the severity of vemurafenib-associated renal impairment. Instead, we found that inhibition of ferrochelatase (FECH), an enzyme involved in heme biosynthesis contributes to vemurafenib nephrotoxicity. FECH overexpression protected RTECs and conversely FECH knockdown increased the sensitivity to vemurafenib nephrotoxicity. Collectively, these studies suggest that vemurafenib-associated RTEC dysfunction and nephrotoxicity is BRAF-independent and caused in part by off-target FECH inhibition. Translational StatementBRAF is the most frequently mutated protein kinase and a critical oncogenic driver in human cancers. In melanoma and other cancers with BRAF activating mutations, BRAF targeted small-molecule therapeutics such as vemurafenib, and dabrafenib have shown remarkable clinical benefits. However, recent clinical studies have shown that a significant number of patients that receive vemurafenib develop AKI through mechanisms that remain unknown. The present study describes the development of novel experimental models of vemurafenib nephrotoxicity and reveals the underlying off-target mechanisms that contribute to renal injury.

pharmacology and toxicology↗

bioRxiv2019

Modeling acute kidney injury in kidney organoids with cisplatin

Acute kidney injury (AKI) remains a major global healthcare problem and there is a need to develop human-based models to study AKI in vitro. Towards this goal, we have characterized induced pluripotent stem cell-derived human kidney organoids and their response to cisplatin, a chemotherapeutic drug that induces AKI and preferentially damages the proximal tubule. We found that a single treatment with 50 {micro}M cisplatin induces HAVCR1 and CXCL8 expression, DNA damage ({gamma}H2AX) and cell death in the organoids in a dose-dependent manner but greatly impairs organoid viability. DNA damage was not specific to the proximal tubule but also affected the distal tubule and interstitial populations. This lack of specificity correlated with low expression of the proximal tubule-specific SLC22A2/OCT2 transporter for cisplatin. To improve viability, we developed a repeated low-dose regimen of 4x 5 {micro}M cisplatin over 7 days and found this causing less toxicity while still inducing a robust AKI response that included secretion of known AKI biomarkers and inflammatory cytokines. This work validates the use of human kidney organoids to model aspects of AKI in vitro, with the potential to identify new AKI biomarkers and develop better therapies.

developmental biology↗

bioRxiv2017

A simple bioreactor-based method to generate kidney organoids from pluripotent stem cells

Kidney organoids generated from human pluripotent stem cells have the potential to revolutionize how kidney development and injury are studied. Current protocols are technically complex and suffer from poor reproducibility and high reagent costs restricting scalability. To overcome these issues, we have established a simple, inexpensive and robust method to grow kidney organoids in bulk from human induced pluripotent stem cells. Our organoids develop tubular structures by day (d) 8 and show optimal tissue morphology at d14. A comparison with fetal human kidney suggests that d14 organoid renal structures most closely resemble capillary loop stage nephrons. We show that deletion of HNF1B, a transcription factor linked to congenital kidney defects, interferes with tubulogenesis, validating our experimental system for studying renal developmental biology. Taken together, our protocol provides a fast, efficient and cost-effective method for generating large quantities of human fetal kidney tissue, enabling the study of normal and aberrant human renal development.

developmental biology↗