title: Visualizing the origins of selfish de novo mutations in individual seminiferous tubules of human testes.
abstract: De novo point mutations arise predominantly in the male germline and increase in frequency with age, but it has not previously been possible to locate specific, identifiable mutations directly within the seminiferous tubules of human testes. Using microdissection of tubules exhibiting altered expression of the spermatogonial markers MAGEA4, FGFR3, and phospho-AKT, whole genome amplification, and DNA sequencing, we establish an in situ strategy for discovery and analysis of pathogenic de novo mutations. In 14 testes from men aged 39-90 y, we identified 11 distinct gain-of-function mutations in five genes (fibroblast growth factor receptors FGFR2 and FGFR3, tyrosine phosphatase PTPN11, and RAS oncogene homologs HRAS and KRAS) from 16 of 22 tubules analyzed; all mutations have known associations with severe diseases, ranging from congenital or perinatal lethal disorders to somatically acquired cancers. These results support proposed selfish selection of spermatogonial mutations affecting growth factor receptor-RAS signaling, highlight its prevalence in older men, and enable direct visualization of the microscopic anatomy of elongated mutant clones.
authors: Clinical Genetics Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, United Kingdom;
authors: Geoffrey J
authors: GJ
authors: Maher
authors: Computational Biology Research Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, United Kingdom;
authors: Simon J
authors: SJ
authors: McGowan
authors: Clinical Genetics Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, United Kingdom;
authors: Eleni
authors: E
authors: Giannoulatou
authors: Department of Cellular Pathology, Oxford University Hospitals NHS Trust, Oxford, OX3 9DU, United Kingdom.
authors: Clare
authors: C
authors: Verrill
authors: Clinical Genetics Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, United Kingdom; andrew.wilkie@imm.ox.ac.uk anne.goriely@imm.ox.ac.uk.
authors: Anne
authors: A
authors: Goriely
authors: Clinical Genetics Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, United Kingdom; andrew.wilkie@imm.ox.ac.uk anne.goriely@imm.ox.ac.uk.
authors: Andrew O M
authors: AO
authors: Wilkie
mesh_terms: D006801:Humans; D008297:Male; D009154:Mutation; D012671:Seminiferous Tubules / Q000378:metabolism*
keywords: germline; mutation; selfish selection; seminiferous tubule; testis