Derivation of induced pluripotent stem cell lines from New Zealand donors

Oh, Jin Kyo; Przepiorski, Aneta; Chang, Hao-Han; Dodd, Rachel C.; Sander, Veronika; Sorrenson, Brie; Shih, Jen‐Hsing; Hollywood, Jennifer A.; Zoysa, Janak R de; Shepherd, Peter R.; Davidson, Alan J.; Holm, Teresa M.

doi:10.1080/03036758.2020.1830808

Cited by 9 publications

(4 citation statements)

References 16 publications

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“…Kidney organoids were generated from MANZ-2 and MANZ-4 iPSC lines as described. ,, Day 12 organoids were distributed into 24-well ultralow attachment plates (Corning; ≥20 per well), and polymyxin analogues (50 mM stocks in H 2 O) were added at 0.1–2 mM to 0.5 mL of the organoid maintenance medium (“stage 2” medium as described). , Kidney tissue deterioration was scored on bright-field images of whole organoids after 48 h of exposure on an EVOS XL inverted microscope. The treatment period was extended from 24 h as in previously undertaken 2D cell toxicity , to allow the compounds to fully penetrate the organoids (∼500 μm diameter spheres).…”

Section: Methodsmentioning

confidence: 99%

Synthesis, Antibacterial Activity, and Nephrotoxicity of Polymyxin B Analogues Modified at Leu-7, d-Phe-6, and the N-Terminus Enabled by S-Lipidation

et al. 2022

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With the post-antibiotic era rapidly approaching, many have turned their attention to developing new treatments, often by structural modification of existing antibiotics. Polymyxins, a family of lipopeptide antibiotics that are used as a last line of defense in the clinic, have recently developed resistance and exhibit significant nephrotoxicity issues. Using thiol–ene chemistry, the facile preparation of six unique S-lipidated building blocks was demonstrated and used to generate lipopeptide mimetics upon incorporation into solid-phase peptide synthesis (SPPS). We then designed and synthesized 38 polymyxin analogues, incorporating these unique building blocks at the N-terminus, or to replace hydrophobic residues at positions 6 and 7 of the native lipopeptides. Several polymyxin analogues bearing one or more S-linked lipids were found to be equipotent to polymyxin, showed minimal kidney nephrotoxicity, and demonstrated activity against several World Health Organisation (WHO) priority pathogens. The S-lipidation strategy has demonstrated potential as a novel approach to prepare innovative new lipopeptide antibiotics.

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Section: Methodsmentioning

confidence: 99%

Synthesis, Antibacterial Activity, and Nephrotoxicity of Polymyxin B Analogues Modified at Leu-7, d-Phe-6, and the N-Terminus Enabled by S-Lipidation

et al. 2022

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“…iPSCs were maintained on 10 cm cell culture dishes coated with Geltrex (Thermo Fisher) or Cultrex (R&D) and mTeSR1 (Stemcell Technologies) medium, passaged every 3–4 days. Experiments were performed with MANZ-2-2 (female) and MANZ4-37 (male) iPSC lines [ 33 ] as stated in the figures.…”

Section: Methodsmentioning

confidence: 99%

Modeling oxidative injury response in human kidney organoids

et al. 2022

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Background Hemolysis occurs in many injury settings and can trigger disease processes. In the kidney, extracellular hemoglobin can induce damage via several mechanisms. These include oxidative stress, mitochondrial dysfunction, and inflammation, which promote fibrosis and chronic kidney disease. Understanding the pathophysiology of these injury pathways offers opportunities to develop new therapeutic strategies. Methods To model hemolysis-induced kidney injury, human kidney organoids were treated with hemin, an iron-containing porphyrin, that generates reactive oxygen species. In addition, we developed an induced pluripotent stem cell line expressing the biosensor, CytochromeC-GFP (CytoC-GFP), which provides a real-time readout of mitochondrial morphology, health, and early apoptotic events. Results We found that hemin-treated kidney organoids show oxidative damage, increased expression of injury markers, impaired functionality of organic anion and cation transport and undergo fibrosis. Injury could be detected in live CytoC-GFP organoids by cytoplasmic localization of fluorescence. Finally, we show that 4-(phenylthio)butanoic acid, an HDAC inhibitor with anti-fibrotic effects in vivo, reduces hemin-induced human kidney organoid fibrosis. Conclusion This work establishes a hemin-induced model of kidney organoid injury. This platform provides a new tool to study the injury and repair response pathways in human kidney tissue and will assist in the development of new therapeutics.

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“…iPSCs were maintained on 10 cm cell culture dishes coated with Geltrex (Thermo Fisher) and mTeSR1 (Stemcell Technologies) medium, and passaged every 3-4 days. Experiments were performed with MANZ-2-2 (female) and MANZ4-37 (male) iPSC lines [32].…”

Section: Methods Ipsc Culturementioning

confidence: 99%

Modeling oxidative injury response in human kidney organoids

Przepiorski

Vanichapol

Espiritu

et al. 2021

Preprint

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BackgroundPersistent acute kidney injury (AKI) leads to tubular atrophy, kidney fibrosis, and, if severe enough, chronic kidney disease (CKD). A common feature of AKI is the generation of excessive reactive oxygen species (ROS) which damage cells and induce inflammation. MethodsHuman kidney organoids were treated with hemin, an iron-containing porphyrin derived from lysed red blood cells, that generates ROS in disease settings such as rhabdomyolysis, sepsis and ischemia reperfusion leading to AKI. In addition, we developed an induced pluripotent stem cell line expressing the biosensor, CytochromeC-GFP (CytoC-GFP), which provides a real-time readout of mitochondrial morphology, health, and early apoptotic events. ResultsWe found that hemin-treated kidney organoids show oxidative damage, increased expression of injury markers, impaired functionality of organic anion and cation transport and undergo fibrosis. Tubule injury could be detected in live CytoC-GFP organoids by cytoplasmic localization of fluorescence. Finally, we show that 4-(phenylthio)butanoic acid, an HDAC inhibitor with anti-fibrotic effects in vivo , reduces hemin-induced human kidney organoid fibrosis. ConclusionTogether this work establishes a hemin-induced model of kidney organoid injury and fibrosis as a new model to study renal repair and a human platform for developing AKI therapeutics.

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Derivation of induced pluripotent stem cell lines from New Zealand donors

Cited by 9 publications

References 16 publications

Synthesis, Antibacterial Activity, and Nephrotoxicity of Polymyxin B Analogues Modified at Leu-7, d-Phe-6, and the N-Terminus Enabled by S-Lipidation

Synthesis, Antibacterial Activity, and Nephrotoxicity of Polymyxin B Analogues Modified at Leu-7, d-Phe-6, and the N-Terminus Enabled by S-Lipidation

Modeling oxidative injury response in human kidney organoids

Modeling oxidative injury response in human kidney organoids

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