Perturbation of ATP6V0A1 as a Driver and Druggable Target for Renal Proximal Tubular Cellular Injury in Nephropathic Cystinosis

Sur, Swastika; Kerwin, Maggie; Pineda, Silvia; Sansanwal, Poonam; Sigdel, Tara K.; Sirota, Marina; Sarwal, Minnie M.

doi:10.1101/2022.05.13.491826

Perturbation of ATP6V0A1 as a Driver and Druggable Target for Renal Proximal Tubular Cellular Injury in Nephropathic Cystinosis

Sur, Swastika
Kerwin, Maggie
Pineda, Silvia
Sansanwal, Poonam
Sigdel, Tara K.
Sirota, Marina
Sarwal, Minnie M.

Revista:

BioRxiv

Año de publicación: 2022

Tipo: Artículo

DOI: 10.1101/2022.05.13.491826 GOOGLE SCHOLAR Acceso abierto editor

Resumen

Understanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to developing new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts isolated from patients with cystinosis were transcriptionally profiled. Lysosomal fractionation, immunoblotting, confocal microscopy, intracellular pH, TEM, mitochondrial stress test, and membrane integrity assays were performed for validation. CRISPR, CTNS -/- RPTECs were generated. Alterations in cell stress, pH, autophagic turnover, and mitochondrial energetics highlighted key changes in the vacuolar (V)-ATPases in patient-derived and CTNS-/- RPTECs. ATP6V0A1 was significantly downregulated in cystinosis and highly co-regulated with loss of CTNS. Correction of ATP6V0A1 rescued cell stress and mitochondrial function. Treatment of CTNS -/- RPTECs with antioxidants astaxanthin (ATX) induced ATP6V0A1 expression and improved autophagosome turnover and mitochondrial integrity.In conclusion, our exploratory transcriptional and in vitro cellular and functional studies confirm that loss of cystinosin in RPTECs, results in a reduction in ATP6V0A1 expression, with changes in intracellular pH, mitochondrial integrity, mitochondrial function, and autophagosome-lysosome clearance. The novel findings are ATP6V0A1’s role in cystinosis-associated renal pathology and among other antioxidants, ATX specifically upregulated ATP6V0A1, improved autophagosome turnover or reduced autophagy and mitochondrial integrity. This is a pilot study highlighting a novel mechanism of tubular injury in cystinosis and requires further study in animal models to clarify its utility in clinical settings.

Referencias bibliográficas

10.1002/(SICI)1098-1004(199912)14:6<454::AID-HUMU2>3.0.CO;2-H
Fuller M , Meikle PJ , Hopwood JJ . Epidemiology of lysosomal storage diseases: an overview. Fabry disease: perspectives from 5 years of FOS: Oxford PharmaGenesis; 2006.
10.1101/gr.10.2.165
10.1542/peds.102.6.e69
(2009), Biochim Biophys Acta, 4, pp. 684
10.1186/s13256-022-03379-7
10.1093/emboj/20.21.5940
10.1038/ng0498-319
10.1093/hmg/8.13.2507
10.1203/00006450-200001000-00007
10.1038/nrneph.2016.182
10.1186/s13023-016-0426-y
10.1007/s00467-010-1627-6
(2021), International journal of molecular sciences, 22
10.1681/ASN.2019070712
10.15252/emmm.202013067
10.1681/ASN.2019090956
10.1159/000495270
10.1007/s10545-016-9919-z
10.1074/mcp.M116.063867
10.1007/s00467-012-2227-4
10.4161/auto.6.7.13099
(2020), Cell death discovery, 6
10.1080/15548627.2018.1446625
10.1128/MCB.00417-13
10.1681/ASN.2009040383
10.1093/hmg/ddt617
10.1681/ASN.2013060577
(2020), JCI insight, 5
10.1021/jf991106k
10.3390/md12010128
10.1016/j.cbpc.2006.12.008
10.3390/md13042105
10.1038/s41598-021-86146-w
10.1186/s12967-015-0388-1
10.1152/ajprenal.1993.265.6.F839
10.1007/BF01152292
10.7326/0003-4819-147-4-200708210-00006
10.1038/s41596-018-0103-9
10.1152/physrev.00045.2003
10.1113/expphysiol.2013.073809
10.1002/emmm.201201527
10.1074/jbc.M116.764076
(2019), Front Endocrinol (Lausanne, 10
10.1159/000331445
10.1086/302509
10.1152/ajprenal.00277.2021
10.1681/ASN.2014090937
10.1056/NEJMra020552
10.1038/ki.2011.277
10.1016/j.neuroscience.2014.01.001
10.1016/j.abb.2008.03.025
10.1126/science.1207056
10.1038/nature11654
10.1091/mbc.E20-06-0383
10.1016/j.ab.2008.10.007
(2017), Frontiers in cell and developmental biology, 5
(2014), Biochim Biophys Acta, 4, pp. 461
10.3389/fnins.2020.00742
10.1172/JCI113331
10.1172/JCI111806
10.3164/jcbn.18-47
10.1093/jn/134.12.3225
(2018), International journal of molecular sciences, 19
10.3892/mmr.2016.5035
10.1016/j.neulet.2017.03.048
(2019), Front Cell Neurosci, 13
10.3390/ijms11125109
10.1016/j.fct.2010.04.002
10.1016/j.intimp.2018.01.011
(2016), Mar Drugs, 14
10.1111/j.1471-4159.2008.05743.x
10.1016/j.brainres.2010.08.100
10.1016/j.ejphar.2017.04.008
10.1007/s12192-013-0443-x
(2017), Mar Drugs, 15
10.1016/j.jpeds.2004.03.056
10.1007/s00467-014-3018-x
10.1203/PDR.0b013e31809fd9a7
10.2165/00003088-197904030-00003
10.1515/JBCPP.1990.1.1-4.357
(2019), 3 Biotech
10.1126/science.157.3794.1321
10.1073/pnas.091062498