Antisense oligonucleotide treatment ameliorates IFN-γ–induced proteinuria in APOL1-transgenic mice

Aghajan, Mariam; Booten, Sheri; Althage, Magnus; Hart, Christopher E.; Ericsson, Anette; Maxvall, Ingela; Ochaba, Joseph; Menschik-Lundin, Angela; Hartleib, Judith; Kuntz, Steven; Gattis, Danielle; Ahlström, Christine; Watt, Andrew T.; Engelhardt, Jeffery A.; Monia, Brett P.; Magnone, Maria Chiara; Guo, Shuling

doi:10.1172/jci.insight.126124

Cited by 77 publications

(93 citation statements)

References 58 publications

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“…RT-qPCR on RNA from whole-well organoids differentiated from G0 and G1 1016SevA lines and the G0 Penn134-61-26 line revealed that organoids expressed little detectable APOL1 under standard culture conditions (Figure 3, A and B, Supplemental Figure 2), consistent with one study showing low levels in human kidneys in vivo (30). IFN-g has been implicated as a factor that may induce APOL1 expression in transgenic mice, although whether this occurs in humans remains unclear (31). Robust APOL1 expression (mean .4000-fold over ACTB, P50.004 by twoway ANOVA) was induced in both G0 and G1 organoids when exposed to 25 ng/ml IFN-g for 24 hours ( Figure 3A).…”

Section: Ifn-g Induces Apol1 Expression In G0 and G1 Kidney Organoidssupporting

confidence: 82%

Profiling APOL1 Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

et al. 2020

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Background DNA variants in APOL1 associate with kidney disease, but the pathophysiologic mechanisms remain incompletely understood. Model organisms lack the APOL1 gene, limiting the degree to which disease states can be recapitulated. Here we present single-cell RNA sequencing (scRNA-seq) of genome-edited human kidney organoids as a platform for profiling effects of APOL1 risk variants in diverse nephron cell types. Methods We performed footprint-free CRISPR-Cas9 genome editing of human induced pluripotent stem cells (iPSCs) to knock in APOL1 high-risk G1 variants at the native genomic locus. iPSCs were differentiated into kidney organoids, treated with vehicle, IFN-g, or the combination of IFN-g and tunicamycin, and analyzed with scRNA-seq to profile cell-specific changes in differential gene expression patterns, compared with isogenic G0 controls. Results Both G0 and G1 iPSCs differentiated into kidney organoids containing nephron-like structures with glomerular epithelial cells, proximal tubules, distal tubules, and endothelial cells. Organoids expressed detectable APOL1 only after exposure to IFN-g. scRNA-seq revealed cell type-specific differences in G1 organoid response to APOL1 induction. Additional stress of tunicamycin exposure led to increased glomerular epithelial cell dedifferentiation in G1 organoids. Conclusions Single-cell transcriptomic profiling of human genome-edited kidney organoids expressing APOL1 risk variants provides a novel platform for studying the pathophysiology of APOL1-mediated kidney disease.

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Section: Ifn-g Induces Apol1 Expression In G0 and G1 Kidney Organoidssupporting

confidence: 82%

Profiling APOL1 Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

et al. 2020

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“…RT-qPCR on RNA from whole-well organoids differentiated from G0 and G1 1016SevA lines and the G0 Penn134-61-26 line revealed that organoids expressed little to no APOL1 under standard culture conditions ( Figure 2A-B, Figure S2). IFN-g has been implicated as a factor that may induce APOL1 expression in mice, although whether this occurs in humans remains unclear 23 . Robust APOL1 expression (mean >4,000-fold over ACTB, P = 0.004 by 2-way ANOVA) was induced in both G0 and G1 organoids when exposed to 25 ng/mL IFN-g for 24 hours (Figure 2A).…”

Section: Resultsmentioning

confidence: 99%

ProfilingAPOL1Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

Liu

Radmanesh

Chung

et al. 2019

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Running Title: APOL1 Variant Organoid scRNA-seq Word Count: 205 (Abstract), 1736 (Main Text, excluding Methods)ABSTRACT Background DNA variants in APOL1 associate with kidney disease, but the pathophysiological mechanisms remain incompletely understood. Model organisms lack the APOL1 gene, limiting the degree to which disease states can be recapitulated. Here we present single-cell RNA sequencing (scRNAseq) of genome-edited human kidney organoids as a platform for profiling effects of APOL1 risk variants in diverse nephron cell types. MethodsWe performed footprint-free CRISPR-Cas9 genome editing of human induced pluripotent stem cells (iPSCs) to knock in APOL1 high-risk G1 variants at the native genomic locus. iPSCs were differentiated into kidney organoids, treated with vehicle, IFN-g, or the combination of IFN-g and tunicamycin, and analyzed with scRNA-seq to profile cell-specific changes in differential gene expression patterns, compared to isogenic G0 controls. ResultsBoth G0 and G1 iPSCs differentiated into kidney organoids containing nephron-like structures with glomerular epithelial cells, proximal tubules, distal tubules, and endothelial cells.Organoids expressed detectable APOL1 only after exposure to IFN-g. scRNA-seq revealed cell type-specific differences in G1 organoid response to APOL1 induction. Additional stress of tunicamycin exposure led to increased glomerular epithelial cell dedifferentiation in G1 organoids. ConclusionsSingle-cell transcriptomic profiling of human genome-edited kidney organoids expressing APOL1 risk variants provides a novel platform for studying the pathophysiology of APOL1mediated kidney disease.

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“…Therapies that target the mRNA products or protein products of the variant alleles are being developed and are in preclinical and early-stage clinical studies. Thus, APOL1 antisense RNA reduces proteinuria in interferonstimulated APOL1 transgenic mice, 8 characteristics, eGFR decline rates were similar between the 2 APOL1 status groups, suggesting that on average, the process of donation may equally stress kidneys of both APOL1 high-risk and low-risk individuals. The worse outcomes in APOL1 highrisk subjects may due to predonation pathophysiology, which is only partly identified by slightly lower eGFR at the time of donation, as eGFR does not provide information about the degree of renal compensation; this is particularly true early in the process of renal compensation for reduced renal mass, when eGFR may be normal.…”

mentioning

confidence: 78%

Antisense oligonucleotide treatment ameliorates IFN-γ–induced proteinuria in APOL1-transgenic mice

Cited by 77 publications

References 58 publications

Profiling APOL1 Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

Profiling APOL1 Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

ProfilingAPOL1Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics

Launching APOLLO: The Role of APOL1 Genetic Variants in Live- and Deceased-Donor Kidney Transplantation

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