Free PMC article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8266742/pdf/nihms-1705620.pdfGEMINI Consortium: Donald F Conrad, Liina Nagirnaja, Kenneth I Aston, Douglas T Carrell, James M Hotaling, Timothy G Jenkins, Rob McLachlan, Moira K O'Bryan, Peter N Schlegel, Michael L Eisenberg, Jay I Sandlow, Emily S Jungheim, Kenan R Omurtag, Alexandra M Lopes, Susana Seixas, Filipa Carvalho, Susana Fernandes, Alberto Barros, João Gonçalves, Iris Caetano, Graça Pinto, Sónia Correia, Maris Laan, Margus Punab, Ewa Rajpert-De Meyts, Niels Jørgensen, Kristian Almstrup, Csilla G Krausz, Keith A Jarvi.Member of GEMINI Consortium: João Gonçalves (INSA), lista completa na pág 1179.Male infertility impacts millions of couples yet, the etiology of primary infertility remains largely unknown. A critical element of successful spermatogenesis is maintenance of genome integrity. Here, we present a genomic study of spermatogenic failure (SPGF). Our initial analysis (n=176) did not reveal known gene-candidates but identifed a potentially signifcant single-nucleotide variant (SNV) in X-linked germ-cell nuclear antigen (GCNA). Together with a larger follow-up study (n=2049), 7 likely clinically relevant GCNA variants were identifed. GCNA is critical for genome integrity in male meiosis and knockout models exhibit impaired spermatogenesis and infertility. Single-cell RNA-seq and immunohistochemistry confrm human GCNA expression from spermatogonia to elongated spermatids. Five identifed SNVs were located in key
functional regions, including N-terminal SUMO-interacting motif and C-terminal Spartan-like protease domain. Notably, variant p.Ala115ProfsTer7 results in an early frameshift, while Spartan-like domain missense variants p.Ser659Trp and p.Arg664Cys change conserved residues, likely afecting 3D structure. For variants within GCNA’s intrinsically disordered region, we performed computational modeling for consensus motifs. Two SNVs were predicted to impact the structure of these consensus motifs. All identifed variants have an extremely low minor allele frequency in the general population and 6 of 7 were not detected in>5000 biological fathers. Considering evidence from animal models, germ-cell-specifc expression, 3D modeling, and computational predictions for SNVs, we propose that identifed GCNA variants disrupt structure and function of the respective protein domains, ultimately arresting germ-cell division. To our knowledge, this is the frst study implicating GCNA, a key genome integrity factor, in human male infertility.This study was supported by The Eunice Kennedy Shriver
NICHD Grant HD080755 (ANY), the Magee-Womens Research
Institute University of Pittsburgh Start Up Fund (ANY), PA DoH
Grant SAP4100085736 (ANY), NIH P50 Specialized Center Grant
HD096723 (KO, ANY, DC, PNS, KH, and MBE), German Research
Foundation Clinical Research Unit ‘Male Germ Cells’ grant DFG
CRU326 (FT), National Science Centre in Poland, grants no.:
2017/26/D/NZ5/00789 (AM) and 2015/17/B/NZ2/01157; NCN
2020/37/B/NZ5/00549 (MK), Magee-Womens Research Institute
University of Pittsburgh, Faculty Fellowship Award and NICHD T32
HD087194 (JH), GM125812 (MB), GM127569 (MB, JLY, and ANY),
NIH R00H090289 (MABE), National Health and Medical Research
Council Project grant APP1120356 (MKOB, JAV, and DC), UUKi
Rutherford Fund Fellowship (BJH), Estonian Research Council, grants
IUT34-12 and PRG1021 (ML), and The Netherlands Organization
for Scientifc Research grant no.: 918-15-667 as well as an Investigator Award in Science from the Wellcome Trust grant no.: 209451 (JAV). Computational analysis was supported in part by the University
of Pittsburgh Center for Research Computing through the resources
provided.info:eu-repo/semantics/publishedVersio
