Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy

Fabian Braun; Ahmed Abed; Dominik Sellung; Manuel Rogg; Mathias Woidy; Oysten Eikrem; Nicola Wanner; Jessica Gambardella; Sandra D. Laufer; Fabian Haas; Milagros N. Wong; Bernhard Dumoulin; Paula Rischke; Anne Mühlig; Wiebke Sachs; Katharina von Cossel; Kristina Schulz; Nicole Muschol; Sören W. Gersting; Ania C. Muntau; Oliver Kretz; Oliver Hahn; Markus M. Rinschen; Michael Mauer; Tillmann Bork; Florian Grahammer; Wei Liang; Thorsten Eierhoff; Winfried Römer; Arne Hansen; Catherine Meyer-Schwesinger; Guido Iaccarino; Camilla Tøndel; Hans Peter Marti; Behzad Najafian; Victor G. Puelles; Christoph Schell; Tobias B. Huber

doi:10.1172/JCI157782

Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy

Fabian Braun, Ahmed Abed, Dominik Sellung, Manuel Rogg, Mathias Woidy, Oysten Eikrem, Nicola Wanner, Jessica Gambardella, Sandra D. Laufer, Fabian Haas, Milagros N. Wong, Bernhard Dumoulin, Paula Rischke, Anne Mühlig, Wiebke Sachs, Katharina von Cossel, Kristina Schulz, Nicole Muschol, Sören W. Gersting, Ania C. MuntauOliver Kretz, Oliver Hahn, Markus M. Rinschen, Michael Mauer, Tillmann Bork, Florian Grahammer, Wei Liang, Thorsten Eierhoff, Winfried Römer, Arne Hansen, Catherine Meyer-Schwesinger, Guido Iaccarino, Camilla Tøndel, Hans Peter Marti, Behzad Najafian, Victor G. Puelles, Christoph Schell, Tobias B. Huber

Pediatrics - Nephrology

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

Current therapies for Fabry disease are based on reversing intracellular accumulation of globotriaosylceramide (Gb3) by enzyme replacement therapy (ERT) or chaperone-mediated stabilization of the defective enzyme, thereby alleviating lysosomal dysfunction. However, their effect in the reversal of end-organ damage, like kidney injury and chronic kidney disease, remains unclear. In this study, ultrastructural analysis of serial human kidney biopsies showed that long-term use of ERT reduced Gb3 accumulation in podocytes but did not reverse podocyte injury. Then, a CRISPR/Cas9-mediated α-galactosidase knockout podocyte cell line confirmed ERT-mediated reversal of Gb3 accumulation without resolution of lysosomal dysfunction. Transcriptome-based connectivity mapping and SILAC-based quantitative proteomics identified α-synuclein (SNCA) accumulation as a key event mediating podocyte injury. Genetic and pharmacological inhibition of SNCA improved lysosomal structure and function in Fabry podocytes, exceeding the benefits of ERT. Together, this work reconceptualizes Fabry-associated cell injury beyond Gb3 accumulation, and introduces SNCA modulation as a potential intervention, especially for patients with Fabry nephropathy.

Original language	English (US)
Article number	e157782
Journal	Journal of Clinical Investigation
Volume	133
Issue number	11
DOIs	https://doi.org/10.1172/JCI157782
State	Published - Jun 1 2023

Bibliographical note

Funding Information:
We thank Charlotte Meyer, Betina Kiefer, Valerie Oberüber, and Anja Obser for expert technical assistance. We thank Ein-ar Svarstad for taking part in the treatment and recruitment of Fabry patients. We also express our gratitude to all members of our laboratories and to the Life Imaging Center of the University of Freiburg for helpful discussions and support. The hR301Q mice of the Naples mouse cohort were provided through Amic- us Therapeutics. This study was supported by the 3R (Replace, Reduce, Refine) Start-up Funding Program, awarded by the Medical Faculty Hamburg in 2018 to FB and AH. FB was supported by the Else Kröner-Fresenius-Stiftung (2021_EKMS.26) and the German Society of Nephrology. DS was supported by Else Kröner-Fresenius-Stiftung grant 2014_MOTIVATE. AM and NW were supported by the Deutsche Forschungsgemein-schaft (DFG; German Research Foundation) (CRC1192). JG is supported by Programma Operativo Nazionale (PON) 1014-2020 REACT-EU. WR acknowledges support by a starting grant from the European Research Council (Programme “Ideas,” ERC-2011-StG 282105), and by the DFG under Germany’s Excellence Strategy (EXC-294 and EXC-2189). CMS was supported by the DFG (ME2108/10-1, INST 152/696-2, SFB1192, project B3). GI is supported by PRIN (2017HTKLRF), from the Italian Ministry of Research. WR and CS were supported by the DFG (SCHE 2092/1-2, SCHE 2092/3-1, SCHE 2092/4-1 [RP9, CP2, CP3] [project IDs 241702976 and 438496892]; CRU329 to CS [project ID 386793560]; CRC1453 [project ID 431984000]; CRC1160 [project ID 256073931]), the Matrix-Code research group (FRIAS, Freiburg), the Excellence Initiative of the German Federal and State Governments, the German Society of Nephrology (DGFN), the Else Kröner-Fresenius-Stiftung, NAKSYS, and the Berta-Ottenstein Programme, Faculty of Medicine, University of Freiburg. TBH was supported by the DFG (CRC1140 [KidGen], CRC1192, HU 1016/8-2, HU 1016/11-1, HU 1016/12-1), the Bundesministerium für Bildung und Forschung — Federal Ministry of Education and Research (BMBF) (STOP-FSGS-01GM2202A, NephrESA-031L0191E, and UPTAKE-01EK2105D), the Else Kröner Fresenius Foundation (Else Kröner-Promotionskolleg, iPRIME), the European Research Council (grant 616891), and the H2020-IMI2 consortium BEAt-DKD (115974); this joint undertaking receives support from the European Union’s Horizon 2020 research and innovation program, the European Federation of Pharmaceutical Industries and Associations, and JDRF. VGP was supported by DFG (CRC1192), the BMBF (STOP-FSGS-01GM2202A and eMed-Consortia Fibromap), and the Novo Nordisk Foundation (Young Investigator Award; NNF21OC0066381).

Publisher Copyright:
Copyright: © 2023, Braun et al. This is an open access article published under the terms of the Creative Commons Attribution 4.0 International License.

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Braun, F., Abed, A., Sellung, D., Rogg, M., Woidy, M., Eikrem, O., Wanner, N., Gambardella, J., Laufer, S. D., Haas, F., Wong, M. N., Dumoulin, B., Rischke, P., Mühlig, A., Sachs, W., von Cossel, K., Schulz, K., Muschol, N., Gersting, S. W., ... Huber, T. B. (2023). Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy. Journal of Clinical Investigation, 133(11), Article e157782. https://doi.org/10.1172/JCI157782

Braun, F, Abed, A, Sellung, D, Rogg, M, Woidy, M, Eikrem, O, Wanner, N, Gambardella, J, Laufer, SD, Haas, F, Wong, MN, Dumoulin, B, Rischke, P, Mühlig, A, Sachs, W, von Cossel, K, Schulz, K, Muschol, N, Gersting, SW, Muntau, AC, Kretz, O, Hahn, O, Rinschen, MM, Mauer, M, Bork, T, Grahammer, F, Liang, W, Eierhoff, T, Römer, W, Hansen, A, Meyer-Schwesinger, C, Iaccarino, G, Tøndel, C, Marti, HP, Najafian, B, Puelles, VG, Schell, C & Huber, TB 2023, 'Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy', Journal of Clinical Investigation, vol. 133, no. 11, e157782. https://doi.org/10.1172/JCI157782

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title = "Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy",

abstract = "Current therapies for Fabry disease are based on reversing intracellular accumulation of globotriaosylceramide (Gb3) by enzyme replacement therapy (ERT) or chaperone-mediated stabilization of the defective enzyme, thereby alleviating lysosomal dysfunction. However, their effect in the reversal of end-organ damage, like kidney injury and chronic kidney disease, remains unclear. In this study, ultrastructural analysis of serial human kidney biopsies showed that long-term use of ERT reduced Gb3 accumulation in podocytes but did not reverse podocyte injury. Then, a CRISPR/Cas9-mediated α-galactosidase knockout podocyte cell line confirmed ERT-mediated reversal of Gb3 accumulation without resolution of lysosomal dysfunction. Transcriptome-based connectivity mapping and SILAC-based quantitative proteomics identified α-synuclein (SNCA) accumulation as a key event mediating podocyte injury. Genetic and pharmacological inhibition of SNCA improved lysosomal structure and function in Fabry podocytes, exceeding the benefits of ERT. Together, this work reconceptualizes Fabry-associated cell injury beyond Gb3 accumulation, and introduces SNCA modulation as a potential intervention, especially for patients with Fabry nephropathy.",

author = "Fabian Braun and Ahmed Abed and Dominik Sellung and Manuel Rogg and Mathias Woidy and Oysten Eikrem and Nicola Wanner and Jessica Gambardella and Laufer, {Sandra D.} and Fabian Haas and Wong, {Milagros N.} and Bernhard Dumoulin and Paula Rischke and Anne M{\"u}hlig and Wiebke Sachs and {von Cossel}, Katharina and Kristina Schulz and Nicole Muschol and Gersting, {S{\"o}ren W.} and Muntau, {Ania C.} and Oliver Kretz and Oliver Hahn and Rinschen, {Markus M.} and Michael Mauer and Tillmann Bork and Florian Grahammer and Wei Liang and Thorsten Eierhoff and Winfried R{\"o}mer and Arne Hansen and Catherine Meyer-Schwesinger and Guido Iaccarino and Camilla T{\o}ndel and Marti, {Hans Peter} and Behzad Najafian and Puelles, {Victor G.} and Christoph Schell and Huber, {Tobias B.}",

note = "Funding Information: We thank Charlotte Meyer, Betina Kiefer, Valerie Ober{\"u}ber, and Anja Obser for expert technical assistance. We thank Ein-ar Svarstad for taking part in the treatment and recruitment of Fabry patients. We also express our gratitude to all members of our laboratories and to the Life Imaging Center of the University of Freiburg for helpful discussions and support. The hR301Q mice of the Naples mouse cohort were provided through Amic- us Therapeutics. This study was supported by the 3R (Replace, Reduce, Refine) Start-up Funding Program, awarded by the Medical Faculty Hamburg in 2018 to FB and AH. FB was supported by the Else Kr{\"o}ner-Fresenius-Stiftung (2021_EKMS.26) and the German Society of Nephrology. DS was supported by Else Kr{\"o}ner-Fresenius-Stiftung grant 2014_MOTIVATE. AM and NW were supported by the Deutsche Forschungsgemein-schaft (DFG; German Research Foundation) (CRC1192). JG is supported by Programma Operativo Nazionale (PON) 1014-2020 REACT-EU. WR acknowledges support by a starting grant from the European Research Council (Programme “Ideas,” ERC-2011-StG 282105), and by the DFG under Germany{\textquoteright}s Excellence Strategy (EXC-294 and EXC-2189). CMS was supported by the DFG (ME2108/10-1, INST 152/696-2, SFB1192, project B3). GI is supported by PRIN (2017HTKLRF), from the Italian Ministry of Research. WR and CS were supported by the DFG (SCHE 2092/1-2, SCHE 2092/3-1, SCHE 2092/4-1 [RP9, CP2, CP3] [project IDs 241702976 and 438496892]; CRU329 to CS [project ID 386793560]; CRC1453 [project ID 431984000]; CRC1160 [project ID 256073931]), the Matrix-Code research group (FRIAS, Freiburg), the Excellence Initiative of the German Federal and State Governments, the German Society of Nephrology (DGFN), the Else Kr{\"o}ner-Fresenius-Stiftung, NAKSYS, and the Berta-Ottenstein Programme, Faculty of Medicine, University of Freiburg. TBH was supported by the DFG (CRC1140 [KidGen], CRC1192, HU 1016/8-2, HU 1016/11-1, HU 1016/12-1), the Bundesministerium f{\"u}r Bildung und Forschung — Federal Ministry of Education and Research (BMBF) (STOP-FSGS-01GM2202A, NephrESA-031L0191E, and UPTAKE-01EK2105D), the Else Kr{\"o}ner Fresenius Foundation (Else Kr{\"o}ner-Promotionskolleg, iPRIME), the European Research Council (grant 616891), and the H2020-IMI2 consortium BEAt-DKD (115974); this joint undertaking receives support from the European Union{\textquoteright}s Horizon 2020 research and innovation program, the European Federation of Pharmaceutical Industries and Associations, and JDRF. VGP was supported by DFG (CRC1192), the BMBF (STOP-FSGS-01GM2202A and eMed-Consortia Fibromap), and the Novo Nordisk Foundation (Young Investigator Award; NNF21OC0066381). Publisher Copyright: Copyright: {\textcopyright} 2023, Braun et al. This is an open access article published under the terms of the Creative Commons Attribution 4.0 International License.",

year = "2023",

month = jun,

day = "1",

doi = "10.1172/JCI157782",

language = "English (US)",

volume = "133",

journal = "Journal of Clinical Investigation",

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TY - JOUR

T1 - Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy

AU - Braun, Fabian

AU - Abed, Ahmed

AU - Sellung, Dominik

AU - Rogg, Manuel

AU - Woidy, Mathias

AU - Eikrem, Oysten

AU - Wanner, Nicola

AU - Gambardella, Jessica

AU - Laufer, Sandra D.

AU - Haas, Fabian

AU - Wong, Milagros N.

AU - Dumoulin, Bernhard

AU - Rischke, Paula

AU - Mühlig, Anne

AU - Sachs, Wiebke

AU - von Cossel, Katharina

AU - Schulz, Kristina

AU - Muschol, Nicole

AU - Gersting, Sören W.

AU - Muntau, Ania C.

AU - Kretz, Oliver

AU - Hahn, Oliver

AU - Rinschen, Markus M.

AU - Mauer, Michael

AU - Bork, Tillmann

AU - Grahammer, Florian

AU - Liang, Wei

AU - Eierhoff, Thorsten

AU - Römer, Winfried

AU - Hansen, Arne

AU - Meyer-Schwesinger, Catherine

AU - Iaccarino, Guido

AU - Tøndel, Camilla

AU - Marti, Hans Peter

AU - Najafian, Behzad

AU - Puelles, Victor G.

AU - Schell, Christoph

AU - Huber, Tobias B.

N1 - Funding Information: We thank Charlotte Meyer, Betina Kiefer, Valerie Oberüber, and Anja Obser for expert technical assistance. We thank Ein-ar Svarstad for taking part in the treatment and recruitment of Fabry patients. We also express our gratitude to all members of our laboratories and to the Life Imaging Center of the University of Freiburg for helpful discussions and support. The hR301Q mice of the Naples mouse cohort were provided through Amic- us Therapeutics. This study was supported by the 3R (Replace, Reduce, Refine) Start-up Funding Program, awarded by the Medical Faculty Hamburg in 2018 to FB and AH. FB was supported by the Else Kröner-Fresenius-Stiftung (2021_EKMS.26) and the German Society of Nephrology. DS was supported by Else Kröner-Fresenius-Stiftung grant 2014_MOTIVATE. AM and NW were supported by the Deutsche Forschungsgemein-schaft (DFG; German Research Foundation) (CRC1192). JG is supported by Programma Operativo Nazionale (PON) 1014-2020 REACT-EU. WR acknowledges support by a starting grant from the European Research Council (Programme “Ideas,” ERC-2011-StG 282105), and by the DFG under Germany’s Excellence Strategy (EXC-294 and EXC-2189). CMS was supported by the DFG (ME2108/10-1, INST 152/696-2, SFB1192, project B3). GI is supported by PRIN (2017HTKLRF), from the Italian Ministry of Research. WR and CS were supported by the DFG (SCHE 2092/1-2, SCHE 2092/3-1, SCHE 2092/4-1 [RP9, CP2, CP3] [project IDs 241702976 and 438496892]; CRU329 to CS [project ID 386793560]; CRC1453 [project ID 431984000]; CRC1160 [project ID 256073931]), the Matrix-Code research group (FRIAS, Freiburg), the Excellence Initiative of the German Federal and State Governments, the German Society of Nephrology (DGFN), the Else Kröner-Fresenius-Stiftung, NAKSYS, and the Berta-Ottenstein Programme, Faculty of Medicine, University of Freiburg. TBH was supported by the DFG (CRC1140 [KidGen], CRC1192, HU 1016/8-2, HU 1016/11-1, HU 1016/12-1), the Bundesministerium für Bildung und Forschung — Federal Ministry of Education and Research (BMBF) (STOP-FSGS-01GM2202A, NephrESA-031L0191E, and UPTAKE-01EK2105D), the Else Kröner Fresenius Foundation (Else Kröner-Promotionskolleg, iPRIME), the European Research Council (grant 616891), and the H2020-IMI2 consortium BEAt-DKD (115974); this joint undertaking receives support from the European Union’s Horizon 2020 research and innovation program, the European Federation of Pharmaceutical Industries and Associations, and JDRF. VGP was supported by DFG (CRC1192), the BMBF (STOP-FSGS-01GM2202A and eMed-Consortia Fibromap), and the Novo Nordisk Foundation (Young Investigator Award; NNF21OC0066381). Publisher Copyright: Copyright: © 2023, Braun et al. This is an open access article published under the terms of the Creative Commons Attribution 4.0 International License.

PY - 2023/6/1

Y1 - 2023/6/1

N2 - Current therapies for Fabry disease are based on reversing intracellular accumulation of globotriaosylceramide (Gb3) by enzyme replacement therapy (ERT) or chaperone-mediated stabilization of the defective enzyme, thereby alleviating lysosomal dysfunction. However, their effect in the reversal of end-organ damage, like kidney injury and chronic kidney disease, remains unclear. In this study, ultrastructural analysis of serial human kidney biopsies showed that long-term use of ERT reduced Gb3 accumulation in podocytes but did not reverse podocyte injury. Then, a CRISPR/Cas9-mediated α-galactosidase knockout podocyte cell line confirmed ERT-mediated reversal of Gb3 accumulation without resolution of lysosomal dysfunction. Transcriptome-based connectivity mapping and SILAC-based quantitative proteomics identified α-synuclein (SNCA) accumulation as a key event mediating podocyte injury. Genetic and pharmacological inhibition of SNCA improved lysosomal structure and function in Fabry podocytes, exceeding the benefits of ERT. Together, this work reconceptualizes Fabry-associated cell injury beyond Gb3 accumulation, and introduces SNCA modulation as a potential intervention, especially for patients with Fabry nephropathy.

AB - Current therapies for Fabry disease are based on reversing intracellular accumulation of globotriaosylceramide (Gb3) by enzyme replacement therapy (ERT) or chaperone-mediated stabilization of the defective enzyme, thereby alleviating lysosomal dysfunction. However, their effect in the reversal of end-organ damage, like kidney injury and chronic kidney disease, remains unclear. In this study, ultrastructural analysis of serial human kidney biopsies showed that long-term use of ERT reduced Gb3 accumulation in podocytes but did not reverse podocyte injury. Then, a CRISPR/Cas9-mediated α-galactosidase knockout podocyte cell line confirmed ERT-mediated reversal of Gb3 accumulation without resolution of lysosomal dysfunction. Transcriptome-based connectivity mapping and SILAC-based quantitative proteomics identified α-synuclein (SNCA) accumulation as a key event mediating podocyte injury. Genetic and pharmacological inhibition of SNCA improved lysosomal structure and function in Fabry podocytes, exceeding the benefits of ERT. Together, this work reconceptualizes Fabry-associated cell injury beyond Gb3 accumulation, and introduces SNCA modulation as a potential intervention, especially for patients with Fabry nephropathy.

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ER -

Accumulation of α-synuclein mediates podocyte injury in Fabry nephropathy

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