175d56d82082b809c2fbad2ed004124c pmc_35052486.pdf 45d10c16d8251102015c0705f9ad19ae40ae31f8 pmc_35052486.pdf c53e181c8366d5d1f08f4befd3420ec78b11eee331b00bac1ea7014aeefcfcb6 pmc_35052486.pdf Title: Gene Network of Susceptibility to Atypical Femoral Fractures Related to Bisphosphonate Treatment Subject: Atypical femoral fractures (AFF) are rare fragility fractures in the subtrocantheric or diaphysis femoral region associated with long-term bisphosphonate (BP) treatment. The etiology of AFF is still unclear even though a genetic basis is suggested. We performed whole exome sequencing (WES) analysis of 12 patients receiving BPs for at least 5 years who sustained AFFs and 4 controls, also long-term treated with BPs but without any fracture. After filtration and prioritization of rare variants predicted to be damaging and present in genes shared among at least two patients, a total of 272 variants in 132 genes were identified. Twelve of these genes were known to be involved in bone metabolism and/or AFF, highlighting DAAM2 and LRP5, both involved in the Wnt pathway, as the most representative. Afterwards, we intersected all mutated genes with a list of 34 genes obtained from a previous study of three sisters with BP-related AFF, identifying nine genes. One of these (MEX3D) harbored damaging variants in two AFF patients from the present study and one shared among the three sisters. Gene interaction analysis using the AFFNET web suggested a complex network among bone-related genes as well as with other mutated genes. BinGO biological function analysis highlighted cytoskeleton and cilium organization. In conclusion, several genes and their interactions could provide genetic susceptibility to AFF, that along with BPs treatment and in some cases with glucocorticoids may trigger this so feared complication. Keywords: atypical femoral fractures; bisphosphonates; WES Author: Natalia Garcia-Giralt, Neus Roca-Ayats, Josep F Abril, Nuria Martinez-Gil, Diana Ovejero, Santos Castañeda, Xavier Nogues, Daniel Grinberg, Susanna Balcells and Raquel Rabionet Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Jan 19 07:16:18 2022 CET ModDate: Wed Jan 19 07:19:05 2022 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2189737 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- VHOBHM+VnURWPalladioL Type 1 Custom yes yes no 10 0 EOWPZV+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 FGHRAU+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 UCLWCM+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 32 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 36 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 40 0 KONDUX+EuphemiaUCAS-Bold Type 1C WinAnsi yes yes no 52 0 HIZBRM+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 55 0 JMPXLZ+URWPalladioL-BoldItal Type 1 Custom yes yes no 77 0 NWYVYM+CMSY10 Type 1 Builtin yes yes no 81 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 93 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 99 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 105 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 04:30:17 CEST RepresentationInformation: pmc_35052486.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2022-01-31 03:38:32 CET Size: 2189737 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 307 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Subjects Destination: subsection.2.1 Item: Title: Whole-Exome Sequencing (WES) Destination: subsection.2.2 Item: Title: AFF Network Construction (AFFNET) Destination: subsection.2.3 Item: Title: Results Destination: section.3 Children: Item: Title: Variant Selection Destination: subsection.3.1 Item: Title: AFF Network Analysis with Candidate Genes Destination: subsection.3.2 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Gene Network of Susceptibility to Atypical Femoral Fractures Related to Bisphosphonate Treatment Author: Natalia Garcia-Giralt, Neus Roca-Ayats, Josep F Abril, Nuria Martinez-Gil, Diana Ovejero, Santos Castañeda, Xavier Nogues, Daniel Grinberg, Susanna Balcells and Raquel Rabionet Subject: Atypical femoral fractures (AFF) are rare fragility fractures in the subtrocantheric or diaphysis femoral region associated with long-term bisphosphonate (BP) treatment. The etiology of AFF is still unclear even though a genetic basis is suggested. We performed whole exome sequencing (WES) analysis of 12 patients receiving BPs for at least 5 years who sustained AFFs and 4 controls, also long-term treated with BPs but without any fracture. After filtration and prioritization of rare variants predicted to be damaging and present in genes shared among at least two patients, a total of 272 variants in 132 genes were identified. Twelve of these genes were known to be involved in bone metabolism and/or AFF, highlighting DAAM2 and LRP5, both involved in the Wnt pathway, as the most representative. Afterwards, we intersected all mutated genes with a list of 34 genes obtained from a previous study of three sisters with BP-related AFF, identifying nine genes. One of these (MEX3D) harbored damaging variants in two AFF patients from the present study and one shared among the three sisters. Gene interaction analysis using the AFFNET web suggested a complex network among bone-related genes as well as with other mutated genes. BinGO biological function analysis highlighted cytoskeleton and cilium organization. In conclusion, several genes and their interactions could provide genetic susceptibility to AFF, that along with BPs treatment and in some cases with glucocorticoids may trigger this so feared complication. 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