Mutations causing medullary cystic kidney disease type 1 lie in a large VNTR in MUC1 missed by massively parallel sequencing
- Creators
- Kirby, Andrew
-
Guttman, Mitchell
Abstract
Although genetic lesions responsible for some mendelian disorders can be rapidly discovered through massively parallel sequencing of whole genomes or exomes, not all diseases readily yield to such efforts. We describe the illustrative case of the simple mendelian disorder medullary cystic kidney disease type 1 (MCKD1), mapped more than a decade ago to a 2-Mb region on chromosome 1. Ultimately, only by cloning, capillary sequencing and de novo assembly did we find that each of six families with MCKD1 harbors an equivalent but apparently independently arising mutation in sequence markedly under-represented in massively parallel sequencing data: the insertion of a single cytosine in one copy (but a different copy in each family) of the repeat unit comprising the extremely long (~1.5–5 kb), GC-rich (>80%) coding variable-number tandem repeat (VNTR) sequence in the MUC1 gene encoding mucin 1. These results provide a cautionary tale about the challenges in identifying the genes responsible for mendelian, let alone more complex, disorders through massively parallel sequencing.
Additional Information
© 2013 Macmillan Publishers Limited. Received 3 May 2012; accepted 7 January 2013; published online 10 February 2013. We thank T.L. Hatte for reagent use. We thank D. Altshuler, T. Carter and J. Schlondorff for useful discussions and M. Cortes, M. Ilzarbe and M. Betancourt for helpful project management. We also thank F. Letendre, M. Coole, R.P. Frere, C. Bonnet, L. Mulrain, N. Norbui and H. Arachchi for Sanger sequencing. This work was conducted as part of the Slim Initiative for Genomic Medicine, a joint United States–Mexico project funded by the Carlos Slim Health Institute. This research was supported in part by the Intramural Research Program of the US NIH, National Human Genome Research Institute (NHGRI). S.K., H.H., J.S. and V.B. were funded by Charles University programs PRVOUK-P24/LF1/3 and UNCE 204011, and their work was supported by grants LH12015 and NT13116-4/2012 from the Ministry of Education and the Ministry of Health of the Czech Republic. S.L.A. was supported by US NIH grant DK34854 (The Harvard Digestive Diseases Center). N.P. is a Broad Fellow of the Broad Institute and a postdoctoral research fellow of the Fund for Scientific Research–Flanders (FWO Vlaanderen, Belgium). I.G.-V. was supported by the Human Frontier Science Program, Alon, the Israeli Centers of Research Excellence (I-CORE) and the Edmond J. Safra Center for Bioinformatics at Tel Aviv University. Author Contributions: A.J.B., E.S.L. and M.J.D. jointly supervised the research. R.J.X., M.R.P. and S.L.A. provided study design and interpretation advice. C.A., S.J.S., P.S.H. and A.J.B. performed sample collection. C. Stevens managed the project. C. Sougnez and K.C. provided early genotyping and sequencing support. Linkage analysis was performed by A.K. on the basis of previous work by P.S.H. A.K. and M.J.D. developed variation discovery and analysis methods. A.K., J.T.R. and R.E.H. analyzed structural variation. T.G. performed CNV analysis. S.G. supervised the sequencing. S. Sigurdsson and K.L.-T. designed the custom capture array. M.P. performed direct PCR of the polymorphic VNTR candidates selected by N.P. A.G. and D.A. performed Southern blot and long-range PCR of the MUC1 VNTR. C.N. supervised the MUC1 VNTR sequencing approach. A.G. performed VNTR allele cloning and generation of sequencing libraries. E.K., R.D., D.P. and S. Steelman performed Sanger sequencing. D.B.J. assembled and analyzed VNTR Sanger sequencing. M.G. provided RNA-seq support. S.K. supervised the immunohistochemistry and immunofluorescence work performed by V.B., H.H., J.S. and P.V. A.K., B.B. and M.D. developed the C-insertion genotype assay. M.C.Z. provided informatic and sequencing consultation. A.R. provided informatic and analysis consultation. C.Y., J.T.R., M.N.C., I.G.-V., R.E.H. and E.R. provided informatic support. The manuscript was written primarily by A.K., A.G., A.J.B., E.S.L. and M.J.D. The supplementary information was prepared mainly by A.K., A.G., D.B.J., B.B., R.E.H., S. Sigurdsson, S.K. and A.J.B. Competing financial interests: A.K., A.G., B.B. and M.D. are listed as inventors on the C-insertion genotyping assay under patent review, filed by the Broad Institute. The other authors declare no competing interests.Attached Files
Accepted Version - nihms474683.pdf
Supplemental Material - ng.2543-S1.pdf
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Additional details
- PMCID
- PMC3901305
- Eprint ID
- 72237
- DOI
- 10.1038/ng.2543
- Resolver ID
- CaltechAUTHORS:20161122-082748151
- Carlos Slim Health Institute
- NIH
- National Human Genome Research Institute
- PRVOUK-P24/LF1/3
- Charles University
- UNCE 204011
- Charles University
- LH12015
- Ministry of Education (Czech Republic)
- NT13116-4/2012
- Ministry of Health (Czech Republic)
- DK34854
- NIH
- Broad Institute of MIT and Harvard
- Fonds Wetenschappelijk Onderzoek - Vlaanderen (FWO)
- Human Frontier Science Program
- Alon Fellowship
- I-CORE Program of the Planning and Budgeting Committee
- Tel Aviv University
- Created
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2016-11-22Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field