Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism
Background: Congenital Hypopituitarism is caused by genetic and environmental factors. Over 30 genes have been implicated in isolated and/or combined pituitary hormone deficiency. The etiology remains unknown for up to 80% of the patients, but most cases have been analyzed by limited candidate gene...
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todo:paper_23249269_v6_n4_p514_PerezMillan2023-10-03T16:40:59Z Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism Pérez Millán, M.I. Vishnopolska, S.A. Daly, A.Z. Bustamante, J.P. Seilicovich, A. Bergadá, I. Braslavsky, D. Keselman, A.C. Lemons, R.M. Mortensen, A.H. Marti, M.A. Camper, S.A. Kitzman, J.O. congenital hypopituitarism GH1 growth hormone deficiency single-molecule molecular inversion probes Background: Congenital Hypopituitarism is caused by genetic and environmental factors. Over 30 genes have been implicated in isolated and/or combined pituitary hormone deficiency. The etiology remains unknown for up to 80% of the patients, but most cases have been analyzed by limited candidate gene screening. Mutations in the PROP1 gene are the most common known cause, and the frequency of mutations in this gene varies greatly by ethnicity. We designed a custom array to assess the frequency of mutations in known hypopituitarism genes and new candidates, using single molecule molecular inversion probes sequencing (smMIPS). Methods: We used this panel for the first systematic screening for causes of hypopituitarism in children. Molecular inversion probes were designed to capture 693 coding exons of 30 known genes and 37 candidate genes. We captured genomic DNA from 51 pediatric patients with CPHD (n = 43) or isolated GH deficiency (IGHD) (n = 8) and their parents and conducted next generation sequencing. Results: We obtained deep coverage over targeted regions and demonstrated accurate variant detection by comparison to whole-genome sequencing in a control individual. We found a dominant mutation GH1, p.R209H, in a three-generation pedigree with IGHD. Conclusions: smMIPS is an efficient and inexpensive method to detect mutations in patients with hypopituitarism, drastically limiting the need for screening individual genes by Sanger sequencing. © 2018 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_23249269_v6_n4_p514_PerezMillan |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
congenital hypopituitarism GH1 growth hormone deficiency single-molecule molecular inversion probes |
| spellingShingle |
congenital hypopituitarism GH1 growth hormone deficiency single-molecule molecular inversion probes Pérez Millán, M.I. Vishnopolska, S.A. Daly, A.Z. Bustamante, J.P. Seilicovich, A. Bergadá, I. Braslavsky, D. Keselman, A.C. Lemons, R.M. Mortensen, A.H. Marti, M.A. Camper, S.A. Kitzman, J.O. Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| topic_facet |
congenital hypopituitarism GH1 growth hormone deficiency single-molecule molecular inversion probes |
| description |
Background: Congenital Hypopituitarism is caused by genetic and environmental factors. Over 30 genes have been implicated in isolated and/or combined pituitary hormone deficiency. The etiology remains unknown for up to 80% of the patients, but most cases have been analyzed by limited candidate gene screening. Mutations in the PROP1 gene are the most common known cause, and the frequency of mutations in this gene varies greatly by ethnicity. We designed a custom array to assess the frequency of mutations in known hypopituitarism genes and new candidates, using single molecule molecular inversion probes sequencing (smMIPS). Methods: We used this panel for the first systematic screening for causes of hypopituitarism in children. Molecular inversion probes were designed to capture 693 coding exons of 30 known genes and 37 candidate genes. We captured genomic DNA from 51 pediatric patients with CPHD (n = 43) or isolated GH deficiency (IGHD) (n = 8) and their parents and conducted next generation sequencing. Results: We obtained deep coverage over targeted regions and demonstrated accurate variant detection by comparison to whole-genome sequencing in a control individual. We found a dominant mutation GH1, p.R209H, in a three-generation pedigree with IGHD. Conclusions: smMIPS is an efficient and inexpensive method to detect mutations in patients with hypopituitarism, drastically limiting the need for screening individual genes by Sanger sequencing. © 2018 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc. |
| format |
JOUR |
| author |
Pérez Millán, M.I. Vishnopolska, S.A. Daly, A.Z. Bustamante, J.P. Seilicovich, A. Bergadá, I. Braslavsky, D. Keselman, A.C. Lemons, R.M. Mortensen, A.H. Marti, M.A. Camper, S.A. Kitzman, J.O. |
| author_facet |
Pérez Millán, M.I. Vishnopolska, S.A. Daly, A.Z. Bustamante, J.P. Seilicovich, A. Bergadá, I. Braslavsky, D. Keselman, A.C. Lemons, R.M. Mortensen, A.H. Marti, M.A. Camper, S.A. Kitzman, J.O. |
| author_sort |
Pérez Millán, M.I. |
| title |
Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| title_short |
Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| title_full |
Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| title_fullStr |
Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| title_full_unstemmed |
Next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| title_sort |
next generation sequencing panel based on single molecule molecular inversion probes for detecting genetic variants in children with hypopituitarism |
| url |
http://hdl.handle.net/20.500.12110/paper_23249269_v6_n4_p514_PerezMillan |
| work_keys_str_mv |
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