Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy

Yo Jun Choi; Jan Halbritter; Daniela A. Braun; Markus Schueler; David Schapiro; John Hoon Rim; Sumeda Nandadasa; Won il Choi; Eugen Widmeier; Shirlee Shril; Friederike Körber; Sidharth K. Sethi; Richard P. Lifton; Bodo B. Beck; Suneel S. Apte; Heon Yung Gee; Friedhelm Hildebrandt

doi:10.1016/j.ajhg.2018.11.003

Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy

Yo Jun Choi, Jan Halbritter, Daniela A. Braun, Markus Schueler, David Schapiro, John Hoon Rim, Sumeda Nandadasa, Won il Choi, Eugen Widmeier, Shirlee Shril, Friederike Körber, Sidharth K. Sethi, Richard P. Lifton, Bodo B. Beck, Suneel S. Apte, Heon Yung Gee, Friedhelm Hildebrandt

Department of Pharmacology

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

18 Citations (Scopus)

Abstract

Nephronophthisis-related ciliopathies (NPHP-RCs) are a group of inherited diseases that are associated with defects in primary cilium structure and function. To identify genes mutated in NPHP-RC, we performed homozygosity mapping and whole-exome sequencing for >100 individuals, some of whom were single affected individuals born to consanguineous parents and some of whom were siblings of indexes who were also affected by NPHP-RC. We then performed high-throughput exon sequencing in a worldwide cohort of 800 additional families affected by NPHP-RC. We identified two ADAMTS9 mutations (c.4575_4576del [p.Gln1525Hisfs^∗60] and c.194C>G [p.Thr65Arg]) that appear to cause NPHP-RC. Although ADAMTS9 is known to be a secreted extracellular metalloproteinase, we found that ADAMTS9 localized near the basal bodies of primary cilia in the cytoplasm. Heterologously expressed wild-type ADAMTS9, in contrast to mutant proteins detected in individuals with NPHP-RC, localized to the vicinity of the basal body. Loss of ADAMTS9 resulted in shortened cilia and defective sonic hedgehog signaling. Knockout of Adamts9 in IMCD3 cells, followed by spheroid induction, resulted in defective lumen formation, which was rescued by an overexpression of wild-type, but not of mutant, ADAMTS9. Knockdown of adamts9 in zebrafish recapitulated NPHP-RC phenotypes, including renal cysts and hydrocephalus. These findings suggest that the identified mutations in ADAMTS9 cause NPHP-RC and that ADAMTS9 is required for the formation and function of primary cilia.

Original language	English
Pages (from-to)	45-54
Number of pages	10
Journal	American Journal of Human Genetics
Volume	104
Issue number	1
DOIs	https://doi.org/10.1016/j.ajhg.2018.11.003
Publication status	Published - 2019 Jan 3

Bibliographical note

Funding Information:
The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health ( DK076683 , DK068306 ). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079 ). S.S.A. is supported by the Allen Distinguished Investigator Program , The Paul G. Allen Frontiers Group , the American Heart Association , and the NIH-NHLBI Program of Excellence in Glycoscience HL107147 . S.N. is supported by the Mark Lauer Pediatric Research Grant . E.W. was supported by the Leopoldina Fellowship Program , German National Academy of Sciences Leopoldina ( LPDS 2015-07 ). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India).

Funding Information:
The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health (DK076683, DK068306). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079). S.S.A. is supported by the Allen Distinguished Investigator Program, The Paul G. Allen Frontiers Group, the American Heart Association, and the NIH-NHLBI Program of Excellence in Glycoscience HL107147. S.N. is supported by the Mark Lauer Pediatric Research Grant. E.W. was supported by the Leopoldina Fellowship Program, German National Academy of Sciences Leopoldina (LPDS 2015-07). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India).

Publisher Copyright:
© 2018 American Society of Human Genetics

All Science Journal Classification (ASJC) codes

Genetics
Genetics(clinical)

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10.1016/j.ajhg.2018.11.003

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Choi, Y. J., Halbritter, J., Braun, D. A., Schueler, M., Schapiro, D., Rim, J. H., Nandadasa, S., Choi, W. I., Widmeier, E., Shril, S., Körber, F., Sethi, S. K., Lifton, R. P., Beck, B. B., Apte, S. S., Gee, H. Y., & Hildebrandt, F. (2019). Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy. American Journal of Human Genetics, 104(1), 45-54. https://doi.org/10.1016/j.ajhg.2018.11.003

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title = "Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy",

abstract = "Nephronophthisis-related ciliopathies (NPHP-RCs) are a group of inherited diseases that are associated with defects in primary cilium structure and function. To identify genes mutated in NPHP-RC, we performed homozygosity mapping and whole-exome sequencing for >100 individuals, some of whom were single affected individuals born to consanguineous parents and some of whom were siblings of indexes who were also affected by NPHP-RC. We then performed high-throughput exon sequencing in a worldwide cohort of 800 additional families affected by NPHP-RC. We identified two ADAMTS9 mutations (c.4575_4576del [p.Gln1525Hisfs∗60] and c.194C>G [p.Thr65Arg]) that appear to cause NPHP-RC. Although ADAMTS9 is known to be a secreted extracellular metalloproteinase, we found that ADAMTS9 localized near the basal bodies of primary cilia in the cytoplasm. Heterologously expressed wild-type ADAMTS9, in contrast to mutant proteins detected in individuals with NPHP-RC, localized to the vicinity of the basal body. Loss of ADAMTS9 resulted in shortened cilia and defective sonic hedgehog signaling. Knockout of Adamts9 in IMCD3 cells, followed by spheroid induction, resulted in defective lumen formation, which was rescued by an overexpression of wild-type, but not of mutant, ADAMTS9. Knockdown of adamts9 in zebrafish recapitulated NPHP-RC phenotypes, including renal cysts and hydrocephalus. These findings suggest that the identified mutations in ADAMTS9 cause NPHP-RC and that ADAMTS9 is required for the formation and function of primary cilia.",

author = "Choi, {Yo Jun} and Jan Halbritter and Braun, {Daniela A.} and Markus Schueler and David Schapiro and Rim, {John Hoon} and Sumeda Nandadasa and Choi, {Won il} and Eugen Widmeier and Shirlee Shril and Friederike K{\"o}rber and Sethi, {Sidharth K.} and Lifton, {Richard P.} and Beck, {Bodo B.} and Apte, {Suneel S.} and Gee, {Heon Yung} and Friedhelm Hildebrandt",

note = "Funding Information: The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health ( DK076683 , DK068306 ). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079 ). S.S.A. is supported by the Allen Distinguished Investigator Program , The Paul G. Allen Frontiers Group , the American Heart Association , and the NIH-NHLBI Program of Excellence in Glycoscience HL107147 . S.N. is supported by the Mark Lauer Pediatric Research Grant . E.W. was supported by the Leopoldina Fellowship Program , German National Academy of Sciences Leopoldina ( LPDS 2015-07 ). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India). Funding Information: The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health (DK076683, DK068306). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079). S.S.A. is supported by the Allen Distinguished Investigator Program, The Paul G. Allen Frontiers Group, the American Heart Association, and the NIH-NHLBI Program of Excellence in Glycoscience HL107147. S.N. is supported by the Mark Lauer Pediatric Research Grant. E.W. was supported by the Leopoldina Fellowship Program, German National Academy of Sciences Leopoldina (LPDS 2015-07). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India). Publisher Copyright: {\textcopyright} 2018 American Society of Human Genetics",

year = "2019",

month = jan,

day = "3",

doi = "10.1016/j.ajhg.2018.11.003",

language = "English",

volume = "104",

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journal = "American Journal of Human Genetics",

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Choi, YJ, Halbritter, J, Braun, DA, Schueler, M, Schapiro, D, Rim, JH, Nandadasa, S, Choi, WI, Widmeier, E, Shril, S, Körber, F, Sethi, SK, Lifton, RP, Beck, BB, Apte, SS, Gee, HY & Hildebrandt, F 2019, 'Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy', American Journal of Human Genetics, vol. 104, no. 1, pp. 45-54. https://doi.org/10.1016/j.ajhg.2018.11.003

TY - JOUR

T1 - Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy

AU - Choi, Yo Jun

AU - Halbritter, Jan

AU - Braun, Daniela A.

AU - Schueler, Markus

AU - Schapiro, David

AU - Rim, John Hoon

AU - Nandadasa, Sumeda

AU - Choi, Won il

AU - Widmeier, Eugen

AU - Shril, Shirlee

AU - Körber, Friederike

AU - Sethi, Sidharth K.

AU - Lifton, Richard P.

AU - Beck, Bodo B.

AU - Apte, Suneel S.

AU - Gee, Heon Yung

AU - Hildebrandt, Friedhelm

N1 - Funding Information: The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health ( DK076683 , DK068306 ). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079 ). S.S.A. is supported by the Allen Distinguished Investigator Program , The Paul G. Allen Frontiers Group , the American Heart Association , and the NIH-NHLBI Program of Excellence in Glycoscience HL107147 . S.N. is supported by the Mark Lauer Pediatric Research Grant . E.W. was supported by the Leopoldina Fellowship Program , German National Academy of Sciences Leopoldina ( LPDS 2015-07 ). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India). Funding Information: The authors thank the families who contributed to this study. F.H. is the Warren E. Grupe professor. F.H. was supported by grants from the National Institutes of Health (DK076683, DK068306). H.Y.G. was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT 2018R1A2A3074572 and 2018R1A5A2025079). S.S.A. is supported by the Allen Distinguished Investigator Program, The Paul G. Allen Frontiers Group, the American Heart Association, and the NIH-NHLBI Program of Excellence in Glycoscience HL107147. S.N. is supported by the Mark Lauer Pediatric Research Grant. E.W. was supported by the Leopoldina Fellowship Program, German National Academy of Sciences Leopoldina (LPDS 2015-07). We thank Yonsei Advanced Imaging Center in cooperation with Carl Zeiss Microscopy and Electron Microscopy Core. We also thank Dr. Seok Jun Moon (Yonsei University College of Dentistry) for sharing the SMO-GFP RPE1 cells, and we thank Drs. Shyam Bihari Bansal and Vijay Kher (Kidney Institute, Medanta, the Medicity, India). Publisher Copyright: © 2018 American Society of Human Genetics

PY - 2019/1/3

Y1 - 2019/1/3

N2 - Nephronophthisis-related ciliopathies (NPHP-RCs) are a group of inherited diseases that are associated with defects in primary cilium structure and function. To identify genes mutated in NPHP-RC, we performed homozygosity mapping and whole-exome sequencing for >100 individuals, some of whom were single affected individuals born to consanguineous parents and some of whom were siblings of indexes who were also affected by NPHP-RC. We then performed high-throughput exon sequencing in a worldwide cohort of 800 additional families affected by NPHP-RC. We identified two ADAMTS9 mutations (c.4575_4576del [p.Gln1525Hisfs∗60] and c.194C>G [p.Thr65Arg]) that appear to cause NPHP-RC. Although ADAMTS9 is known to be a secreted extracellular metalloproteinase, we found that ADAMTS9 localized near the basal bodies of primary cilia in the cytoplasm. Heterologously expressed wild-type ADAMTS9, in contrast to mutant proteins detected in individuals with NPHP-RC, localized to the vicinity of the basal body. Loss of ADAMTS9 resulted in shortened cilia and defective sonic hedgehog signaling. Knockout of Adamts9 in IMCD3 cells, followed by spheroid induction, resulted in defective lumen formation, which was rescued by an overexpression of wild-type, but not of mutant, ADAMTS9. Knockdown of adamts9 in zebrafish recapitulated NPHP-RC phenotypes, including renal cysts and hydrocephalus. These findings suggest that the identified mutations in ADAMTS9 cause NPHP-RC and that ADAMTS9 is required for the formation and function of primary cilia.

AB - Nephronophthisis-related ciliopathies (NPHP-RCs) are a group of inherited diseases that are associated with defects in primary cilium structure and function. To identify genes mutated in NPHP-RC, we performed homozygosity mapping and whole-exome sequencing for >100 individuals, some of whom were single affected individuals born to consanguineous parents and some of whom were siblings of indexes who were also affected by NPHP-RC. We then performed high-throughput exon sequencing in a worldwide cohort of 800 additional families affected by NPHP-RC. We identified two ADAMTS9 mutations (c.4575_4576del [p.Gln1525Hisfs∗60] and c.194C>G [p.Thr65Arg]) that appear to cause NPHP-RC. Although ADAMTS9 is known to be a secreted extracellular metalloproteinase, we found that ADAMTS9 localized near the basal bodies of primary cilia in the cytoplasm. Heterologously expressed wild-type ADAMTS9, in contrast to mutant proteins detected in individuals with NPHP-RC, localized to the vicinity of the basal body. Loss of ADAMTS9 resulted in shortened cilia and defective sonic hedgehog signaling. Knockout of Adamts9 in IMCD3 cells, followed by spheroid induction, resulted in defective lumen formation, which was rescued by an overexpression of wild-type, but not of mutant, ADAMTS9. Knockdown of adamts9 in zebrafish recapitulated NPHP-RC phenotypes, including renal cysts and hydrocephalus. These findings suggest that the identified mutations in ADAMTS9 cause NPHP-RC and that ADAMTS9 is required for the formation and function of primary cilia.

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ER -

Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy

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