Induced Pluripotent Stem Cell–Derived Podocyte-Like Cells as Models for Assessing Mechanisms Underlying Heritable Disease Phenotype: Initial Studies Using Two Alport Syndrome Patient Lines Indicate Impaired Potassium Channel Activity
| Autor: | Wai-Ling Lieuw, Teresa H. Vandekolk, Sharon D. Ricardo, John M. Haynes, James N. Selby, Sheetal Saini, Joan K. Ho, Craig L. Fisher, Katie Leach, Isaiah P. L. Abad, Judith Savige |
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| Rok vydání: | 2018 |
| Předmět: |
Collagen Type IV
Male 0301 basic medicine Potassium Channels Adolescent Induced Pluripotent Stem Cells Nephritis Hereditary Calcium in biology Cell Line Podocyte 03 medical and health sciences 0302 clinical medicine Glomerular Basement Membrane medicine Humans Alport syndrome Large-Conductance Calcium-Activated Potassium Channel alpha Subunits Induced pluripotent stem cell Pharmacology Mesangial cell Podocytes Chemistry Glomerular basement membrane Middle Aged medicine.disease Cell biology Potassium channel activity Phenotype 030104 developmental biology medicine.anatomical_structure Focal Adhesion Protein-Tyrosine Kinases Molecular Medicine Calcium Stem cell Receptors Calcium-Sensing 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | Journal of Pharmacology and Experimental Therapeutics. 367:335-347 |
| ISSN: | 1521-0103 0022-3565 |
| DOI: | 10.1124/jpet.118.250142 |
| Popis: | Renal podocyte survival depends upon the dynamic regulation of a complex cell architecture that links the glomerular basement membrane to integrins, ion channels, and receptors. Alport syndrome is a heritable chronic kidney disease where mutations in α3, α4, or α5 collagen genes promote podocyte death. In rodent models of renal failure, activation of the calcium-sensing receptor (CaSR) can protect podocytes from stress-related death. In this study, we assessed CaSR function in podocyte-like cells derived from induced-pluripotent stem cells from two patients with Alport Syndrome (AS1 & AS2) and a renal disease free individual [normal human mesangial cell (NHMC)], as well as a human immortalized podocyte-like (HIP) cell line. Extracellular calcium elicited concentration-dependent elevations of intracellular calcium in all podocyte-like cells. NHMC and HIP, but not AS1 or AS2 podocyte-like cells, also showed acute reductions in intracellular calcium prior to elevation. In NHMC podocyte-like cells this acute reduction was blocked by the large-conductance potassium channel (KCNMA1) inhibitors iberiotoxin (10 nM) and tetraethylammonium (5 mM), as well as the focal adhesion kinase inhibitor PF562271 (N-methyl-N-(3-((2-(2-oxo-2,3-dihydro-1H-indol-5-ylamino)-5-trifluoromethyl-pyrimidin-4-ylamino)-methyl)-pyridin-2-yl)-methanesulfonamide, 10 nM). Quantitative polymerase chain reaction (qPCR) and immunolabeling showed the presence of KCNMA1 transcript and protein in all podocyte-like cells tested. Cultivation of AS1 podocytes on decellularized plates of NHMC podocyte-like cells partially restored acute reductions in intracellular calcium in response to extracellular calcium. We conclude that the AS patient-derived podocyte-like cells used in this study showed dysfunctional integrin signaling and potassium channel function, which may contribute to podocyte death seen in Alport syndrome. |
| Databáze: | OpenAIRE |
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