Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome

Youying Mao; Ronen Schneider; Peter F.M. van der Ven; Marvin Assent; Keerthika Lohanadan; Verena Klämbt; Florian Buerger; Thomas M. Kitzler; Konstantin Deutsch; Makiko Nakayama; Amar J. Majmundar; Nina Mann; Tobias Hermle; Ana C. Onuchic-Whitford; Wei Zhou; Nandini Nagarajan Margam; Roy Duncan; Jonathan Marquez; Mustafa Khokha; Hanan M. Fathy; Jameela A. Kari; Sherif El Desoky; Loai A. Eid; Hazem Subhi Awad; Muna Al-Saffar; Shrikant Mane; Richard P. Lifton; Dieter O. Fürst; Shirlee Shril; Friedhelm Hildebrandt

doi:10.1016/j.ekir.2020.10.040

Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome

Youying Mao, Ronen Schneider, Peter F.M. van der Ven, Marvin Assent, Keerthika Lohanadan, Verena Klämbt, Florian Buerger, Thomas M. Kitzler, Konstantin Deutsch, Makiko Nakayama, Amar J. Majmundar, Nina Mann, Tobias Hermle, Ana C. Onuchic-Whitford, Wei Zhou, Nandini Nagarajan Margam, Roy Duncan, Jonathan Marquez, Mustafa Khokha, Hanan M. FathyJameela A. Kari, Sherif El Desoky, Loai A. Eid, Hazem Subhi Awad, Muna Al-Saffar, Shrikant Mane, Richard P. Lifton, Dieter O. Fürst, Shirlee Shril, Friedhelm Hildebrandt

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

7 Citas (Scopus)

Resumen

Introduction: Most of the approximately 60 genes that if mutated cause steroid-resistant nephrotic syndrome (SRNS) are highly expressed in the glomerular podocyte, rendering SRNS a “podocytopathy.” Methods: We performed whole-exome sequencing (WES) in 1200 nephrotic syndrome (NS) patients. Results: We discovered homozygous truncating and homozygous missense mutation in SYNPO2 (synaptopodin-2) (p.Lys1124∗ and p.Ala1134Thr) in 2 patients with childhood-onset NS. We found SYNPO2 expression in both podocytes and mesangial cells; however, notably, immunofluorescence staining of adult human and rat kidney cryosections indicated that SYNPO2 is localized mainly in mesangial cells. Subcellular localization studies reveal that in these cells SYNPO2 partially co-localizes with α-actinin and filamin A−containing F-actin filaments. Upon transfection in mesangial cells or podocytes, EGFP-SYNPO2 co-localized with α-actinin-4, which gene is mutated in autosomal dominant SRNS in humans. SYNPO2 overexpression increases mesangial cell migration rate (MMR), whereas shRNA knockdown reduces MMR. Decreased MMR was rescued by transfection of wild-type mouse Synpo2 cDNA but only partially by cDNA representing mutations from the NS patients. The increased mesangial cell migration rate (MMR) by SYNPO2 overexpression was inhibited by ARP complex inhibitor CK666. SYNPO2 shRNA knockdown in podocytes decreased active Rac1, which was rescued by transfection of wild-type SYNPO2 cDNA but not by cDNA representing any of the 2 mutant variants. Conclusion: We show that SYNPO2 variants may lead to Rac1-ARP3 dysregulation, and may play a role in the pathogenesis of nephrotic syndrome.

Idioma original	English
Páginas (desde-hasta)	472-483
Número de páginas	12
Publicación	Kidney International Reports
Volumen	6
N.º	2
DOI	https://doi.org/10.1016/j.ekir.2020.10.040
Estado	Published - feb. 2021

Nota bibliográfica

Funding Information:
We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH ( 5R01DK076683-13 ) and by the “EPT” program of Shanghai Children’s Medical Center to YM. AJM was supported by an NIH Training Grant ( T32DK-007726 ), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute , and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant ( 404527522 ) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award ( DK122766 ).

Funding Information:
We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH (5R01DK076683-13) and by the “EPT” program of Shanghai Children's Medical Center to YM. AJM was supported by an NIH Training Grant (T32DK-007726), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute, and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant (404527522) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award (DK122766). YM, RS, VK, FB, TMK, NM, MN, ACO-W, AJM, TH, KD, WZ, SS, and FH performed genetic analysis and functional studies. YM and RS performed cDNA cloning, IFs, migration assay, and G-LISA. KL, MA, PFMvdV, and DOF performed GST-Pulldown assay, RT-PCR, transfections, and IF in kidney. NNM and RD performed lamellipodia formation. JM and MK performed in situ hybridizaion experiments in Xenopus. HMF recruited patients and gathered detailed clinical information for the study. SM and RPL performed WES. FH conceived of and directed the project. YM, RS, and FH wrote the paper, which was critically reviewed by all authors. HMF, JAK, SED, LAE, HSA, and MA-S contributed patient data.

Publisher Copyright:
© 2020 International Society of Nephrology

ASJC Scopus Subject Areas

Nephrology

PubMed: MeSH publication types

Journal Article

Acceder al documento

10.1016/j.ekir.2020.10.040

Otros archivos y enlaces

Citar esto

Mao, Y., Schneider, R., van der Ven, P. F. M., Assent, M., Lohanadan, K., Klämbt, V., Buerger, F., Kitzler, T. M., Deutsch, K., Nakayama, M., Majmundar, A. J., Mann, N., Hermle, T., Onuchic-Whitford, A. C., Zhou, W., Margam, N. N., Duncan, R., Marquez, J., Khokha, M., ... Hildebrandt, F. (2021). Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome. Kidney International Reports, 6(2), 472-483. https://doi.org/10.1016/j.ekir.2020.10.040

Mao, Y, Schneider, R, van der Ven, PFM, Assent, M, Lohanadan, K, Klämbt, V, Buerger, F, Kitzler, TM, Deutsch, K, Nakayama, M, Majmundar, AJ, Mann, N, Hermle, T, Onuchic-Whitford, AC, Zhou, W, Margam, NN, Duncan, R, Marquez, J, Khokha, M, Fathy, HM, Kari, JA, El Desoky, S, Eid, LA, Awad, HS, Al-Saffar, M, Mane, S, Lifton, RP, Fürst, DO, Shril, S & Hildebrandt, F 2021, 'Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome', Kidney International Reports, vol. 6, n.º 2, pp. 472-483. https://doi.org/10.1016/j.ekir.2020.10.040

@article{737179bf2baa420a91538346cdf558a3,

title = "Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome",

abstract = "Introduction: Most of the approximately 60 genes that if mutated cause steroid-resistant nephrotic syndrome (SRNS) are highly expressed in the glomerular podocyte, rendering SRNS a “podocytopathy.” Methods: We performed whole-exome sequencing (WES) in 1200 nephrotic syndrome (NS) patients. Results: We discovered homozygous truncating and homozygous missense mutation in SYNPO2 (synaptopodin-2) (p.Lys1124∗ and p.Ala1134Thr) in 2 patients with childhood-onset NS. We found SYNPO2 expression in both podocytes and mesangial cells; however, notably, immunofluorescence staining of adult human and rat kidney cryosections indicated that SYNPO2 is localized mainly in mesangial cells. Subcellular localization studies reveal that in these cells SYNPO2 partially co-localizes with α-actinin and filamin A−containing F-actin filaments. Upon transfection in mesangial cells or podocytes, EGFP-SYNPO2 co-localized with α-actinin-4, which gene is mutated in autosomal dominant SRNS in humans. SYNPO2 overexpression increases mesangial cell migration rate (MMR), whereas shRNA knockdown reduces MMR. Decreased MMR was rescued by transfection of wild-type mouse Synpo2 cDNA but only partially by cDNA representing mutations from the NS patients. The increased mesangial cell migration rate (MMR) by SYNPO2 overexpression was inhibited by ARP complex inhibitor CK666. SYNPO2 shRNA knockdown in podocytes decreased active Rac1, which was rescued by transfection of wild-type SYNPO2 cDNA but not by cDNA representing any of the 2 mutant variants. Conclusion: We show that SYNPO2 variants may lead to Rac1-ARP3 dysregulation, and may play a role in the pathogenesis of nephrotic syndrome.",

author = "Youying Mao and Ronen Schneider and {van der Ven}, {Peter F.M.} and Marvin Assent and Keerthika Lohanadan and Verena Kl{\"a}mbt and Florian Buerger and Kitzler, {Thomas M.} and Konstantin Deutsch and Makiko Nakayama and Majmundar, {Amar J.} and Nina Mann and Tobias Hermle and Onuchic-Whitford, {Ana C.} and Wei Zhou and Margam, {Nandini Nagarajan} and Roy Duncan and Jonathan Marquez and Mustafa Khokha and Fathy, {Hanan M.} and Kari, {Jameela A.} and {El Desoky}, Sherif and Eid, {Loai A.} and Awad, {Hazem Subhi} and Muna Al-Saffar and Shrikant Mane and Lifton, {Richard P.} and F{\"u}rst, {Dieter O.} and Shirlee Shril and Friedhelm Hildebrandt",

note = "Funding Information: We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH ( 5R01DK076683-13 ) and by the “EPT” program of Shanghai Children{\textquoteright}s Medical Center to YM. AJM was supported by an NIH Training Grant ( T32DK-007726 ), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute , and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant ( 404527522 ) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award ( DK122766 ). Funding Information: We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH (5R01DK076683-13) and by the “EPT” program of Shanghai Children's Medical Center to YM. AJM was supported by an NIH Training Grant (T32DK-007726), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute, and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant (404527522) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award (DK122766). YM, RS, VK, FB, TMK, NM, MN, ACO-W, AJM, TH, KD, WZ, SS, and FH performed genetic analysis and functional studies. YM and RS performed cDNA cloning, IFs, migration assay, and G-LISA. KL, MA, PFMvdV, and DOF performed GST-Pulldown assay, RT-PCR, transfections, and IF in kidney. NNM and RD performed lamellipodia formation. JM and MK performed in situ hybridizaion experiments in Xenopus. HMF recruited patients and gathered detailed clinical information for the study. SM and RPL performed WES. FH conceived of and directed the project. YM, RS, and FH wrote the paper, which was critically reviewed by all authors. HMF, JAK, SED, LAE, HSA, and MA-S contributed patient data. Publisher Copyright: {\textcopyright} 2020 International Society of Nephrology",

year = "2021",

month = feb,

doi = "10.1016/j.ekir.2020.10.040",

language = "English",

volume = "6",

pages = "472--483",

journal = "Kidney International Reports",

issn = "2468-0249",

publisher = "Elsevier Inc.",

number = "2",

}

TY - JOUR

T1 - Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome

AU - Mao, Youying

AU - Schneider, Ronen

AU - van der Ven, Peter F.M.

AU - Assent, Marvin

AU - Lohanadan, Keerthika

AU - Klämbt, Verena

AU - Buerger, Florian

AU - Kitzler, Thomas M.

AU - Deutsch, Konstantin

AU - Nakayama, Makiko

AU - Majmundar, Amar J.

AU - Mann, Nina

AU - Hermle, Tobias

AU - Onuchic-Whitford, Ana C.

AU - Zhou, Wei

AU - Margam, Nandini Nagarajan

AU - Duncan, Roy

AU - Marquez, Jonathan

AU - Khokha, Mustafa

AU - Fathy, Hanan M.

AU - Kari, Jameela A.

AU - El Desoky, Sherif

AU - Eid, Loai A.

AU - Awad, Hazem Subhi

AU - Al-Saffar, Muna

AU - Mane, Shrikant

AU - Lifton, Richard P.

AU - Fürst, Dieter O.

AU - Shril, Shirlee

AU - Hildebrandt, Friedhelm

N1 - Funding Information: We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH ( 5R01DK076683-13 ) and by the “EPT” program of Shanghai Children’s Medical Center to YM. AJM was supported by an NIH Training Grant ( T32DK-007726 ), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute , and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant ( 404527522 ) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award ( DK122766 ). Funding Information: We are grateful to the families and study individuals for their contribution. FH is the William E. Harmon Professor of Pediatrics. This research is supported by a grant from the National Institutes of Health to FH (5R01DK076683-13) and by the “EPT” program of Shanghai Children's Medical Center to YM. AJM was supported by an NIH Training Grant (T32DK-007726), the 2017 Post-doctoral Fellowship Grant from the Harvard Stem Cell Institute, and the American Society of Nephrology Lipps Research Program 2018 Polycystic Kidney Disease Foundation Jared J. Grantham Research Fellowship. FB was supported by a fellowship grant (404527522) from the German Research Foundation (DFG). ACO is supported by the National Institutes of Health F32 Ruth L. Kirschstein Postdoctoral Individual National Research Service Award (DK122766). YM, RS, VK, FB, TMK, NM, MN, ACO-W, AJM, TH, KD, WZ, SS, and FH performed genetic analysis and functional studies. YM and RS performed cDNA cloning, IFs, migration assay, and G-LISA. KL, MA, PFMvdV, and DOF performed GST-Pulldown assay, RT-PCR, transfections, and IF in kidney. NNM and RD performed lamellipodia formation. JM and MK performed in situ hybridizaion experiments in Xenopus. HMF recruited patients and gathered detailed clinical information for the study. SM and RPL performed WES. FH conceived of and directed the project. YM, RS, and FH wrote the paper, which was critically reviewed by all authors. HMF, JAK, SED, LAE, HSA, and MA-S contributed patient data. Publisher Copyright: © 2020 International Society of Nephrology

PY - 2021/2

Y1 - 2021/2

N2 - Introduction: Most of the approximately 60 genes that if mutated cause steroid-resistant nephrotic syndrome (SRNS) are highly expressed in the glomerular podocyte, rendering SRNS a “podocytopathy.” Methods: We performed whole-exome sequencing (WES) in 1200 nephrotic syndrome (NS) patients. Results: We discovered homozygous truncating and homozygous missense mutation in SYNPO2 (synaptopodin-2) (p.Lys1124∗ and p.Ala1134Thr) in 2 patients with childhood-onset NS. We found SYNPO2 expression in both podocytes and mesangial cells; however, notably, immunofluorescence staining of adult human and rat kidney cryosections indicated that SYNPO2 is localized mainly in mesangial cells. Subcellular localization studies reveal that in these cells SYNPO2 partially co-localizes with α-actinin and filamin A−containing F-actin filaments. Upon transfection in mesangial cells or podocytes, EGFP-SYNPO2 co-localized with α-actinin-4, which gene is mutated in autosomal dominant SRNS in humans. SYNPO2 overexpression increases mesangial cell migration rate (MMR), whereas shRNA knockdown reduces MMR. Decreased MMR was rescued by transfection of wild-type mouse Synpo2 cDNA but only partially by cDNA representing mutations from the NS patients. The increased mesangial cell migration rate (MMR) by SYNPO2 overexpression was inhibited by ARP complex inhibitor CK666. SYNPO2 shRNA knockdown in podocytes decreased active Rac1, which was rescued by transfection of wild-type SYNPO2 cDNA but not by cDNA representing any of the 2 mutant variants. Conclusion: We show that SYNPO2 variants may lead to Rac1-ARP3 dysregulation, and may play a role in the pathogenesis of nephrotic syndrome.

AB - Introduction: Most of the approximately 60 genes that if mutated cause steroid-resistant nephrotic syndrome (SRNS) are highly expressed in the glomerular podocyte, rendering SRNS a “podocytopathy.” Methods: We performed whole-exome sequencing (WES) in 1200 nephrotic syndrome (NS) patients. Results: We discovered homozygous truncating and homozygous missense mutation in SYNPO2 (synaptopodin-2) (p.Lys1124∗ and p.Ala1134Thr) in 2 patients with childhood-onset NS. We found SYNPO2 expression in both podocytes and mesangial cells; however, notably, immunofluorescence staining of adult human and rat kidney cryosections indicated that SYNPO2 is localized mainly in mesangial cells. Subcellular localization studies reveal that in these cells SYNPO2 partially co-localizes with α-actinin and filamin A−containing F-actin filaments. Upon transfection in mesangial cells or podocytes, EGFP-SYNPO2 co-localized with α-actinin-4, which gene is mutated in autosomal dominant SRNS in humans. SYNPO2 overexpression increases mesangial cell migration rate (MMR), whereas shRNA knockdown reduces MMR. Decreased MMR was rescued by transfection of wild-type mouse Synpo2 cDNA but only partially by cDNA representing mutations from the NS patients. The increased mesangial cell migration rate (MMR) by SYNPO2 overexpression was inhibited by ARP complex inhibitor CK666. SYNPO2 shRNA knockdown in podocytes decreased active Rac1, which was rescued by transfection of wild-type SYNPO2 cDNA but not by cDNA representing any of the 2 mutant variants. Conclusion: We show that SYNPO2 variants may lead to Rac1-ARP3 dysregulation, and may play a role in the pathogenesis of nephrotic syndrome.

UR - http://www.scopus.com/inward/record.url?scp=85099563792&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85099563792&partnerID=8YFLogxK

U2 - 10.1016/j.ekir.2020.10.040

DO - 10.1016/j.ekir.2020.10.040

M3 - Article

C2 - 33615072

AN - SCOPUS:85099563792

SN - 2468-0249

VL - 6

SP - 472

EP - 483

JO - Kidney International Reports

JF - Kidney International Reports

IS - 2

ER -

Recessive Mutations in SYNPO2 as a Candidate of Monogenic Nephrotic Syndrome

Resumen

Nota bibliográfica

ASJC Scopus Subject Areas

PubMed: MeSH publication types

Acceder al documento

Otros archivos y enlaces

Huella

Citar esto