4b8550c186f527b09a852778588e946e 329521.pdf 7a7ab8138fa3f25352623071cecd941d966bc484 329521.pdf dcdfc6e19c9a0c6634f49d5038b7f32959f735bd2941aea0c6565bb3609066d9 329521.pdf Title: VEGF Signaling Pathway Germline Polymorphisms as Prognostic Pharmacogenetic Biomarkers in Localized High-Grade Osteosarcoma Patients from the GEIS-33 Protocol Subject: Background/Objective: The vascular endothelial growth factor (VEGF) signaling pathway induces angiogenesis, which impacts tumor progression and clinical outcomes in patients with localized osteosarcoma. This study evaluates whether genetic polymorphisms in the VEGF signaling pathway are associated with survival outcomes in these patients. Methods: Sixty-nine patients with localized high-grade osteosarcoma enrolled in the GEIS-33 protocol and treated with MAP (methotrexate, doxorubicin, cisplatin) chemotherapy, surgery, and subsequent adjuvant treatment were included. Nine variants of interest in the VEGFA (rs1570360, rs2010963 and rs699947), FLT1 (VEGFR1; rs7993418, rs9513070 and rs9582036), and KDR (VEGFR2; rs1551641 and rs1870377 and rs2071559) genes were genotyped from peripheral blood samples using TaqMan Assay technology. Genetic data were correlated with relapse-free survival (RFS) and overall survival (OS) considering clinical variables as covariates. Results: The analyses showed nominally significant associations between the FLT1 variants rs7993418 and rs9582036 and survival. Patients carrying the rs7993418(C) allele had worse RFS (p = 0.01) and OS (p = 0.01). Carriers of the minor rs9582036(C) allele also had worse RFS (p = 0.02) and OS (p = 0.03). Additionally, patients harboring the TT genotype of the KDR rs1551641 variant had significantly worse RFS (p = 0.002). These polymorphisms remained statistically significant in the multivariate Cox regression analyses that included surgical margins and pathological response as covariates. Conclusions: Pharmacogenetics may contribute to precision medicine in oncology. Germline polymorphisms in the VEGF pathway may be useful as predictors of survival in high-grade localized osteosarcoma patients treated with chemotherapy, following validation in a large cohort of patients. Current treatment strategies aimed at improving outcomes for osteosarcoma patients may benefit from the identification of new biomarkers, such as these FLT1 rs7993418 and rs9582036 variants. Keywords: pharmacogenetics; osteosarcoma; VEGF; polymorphism Author: Juliana Salazar, María J. Arranz, Javier Martin-Broto, Massimo Serra, Emanuela Palmerini, Jeronimo Martínez-García, Antonio Juan-Ribelles, Maitane Andion, Javier Martinez-Trufero, Guiomar Gutiérrez, Aizpea Echebarria-Barona, Maria A. Vaz-Salgado, Alexandra Regueiro, Rosa Alvarez, Claudia Valverde, Pablo Luna, Pilar Blay and Ana Sebio Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Dec 5 09:16:43 2025 CET ModDate: Fri Dec 5 09:31:41 2025 CET Custom Metadata: yes 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: 980851 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PLOLAK+VnURWPalladioL Type 1 Custom yes yes yes 102 0 ECAGAF+URWPalladioL-Ital Type 1 Custom yes yes yes 103 0 BOLNEH+URWPalladioL-Bold Type 1 Custom yes yes yes 104 0 BJKMVU+URWPalladioL-Roma Type 1 Custom yes yes yes 105 0 AEIEME+CMSY10 Type 1 Builtin yes yes yes 190 0 MOMEPI+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 179 0 MOMEPJ+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 180 0 MOMEOH+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 181 0 MOMENF+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 182 0 MOMEOG+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 183 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 237 0 MOMEPI+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 226 0 MOMEPJ+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 227 0 MOMEOH+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 228 0 MOMENF+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 229 0 MOMEOG+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 230 0 LDSSDS+EURM10 Type 1 Builtin yes yes yes 343 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-07-09 04:17:47 CEST RepresentationInformation: 329521.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-07-08 13:08:51 CEST Size: 980851 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 457 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Results Children: Item: Title: Clinical Results Item: Title: VEGF Pathway Polymorphisms and Recurrence-Free Survival Item: Title: VEGF Pathway Polymorphisms and Overall Survival Item: Title: Cumulative Effects of FLT1 rs7993418 and rs9582036 Variants on Survival Item: Title: VEGF Pathway Polymorphisms and Chemotherapy Response Item: Title: Discussion Item: Title: Materials and Methods Children: Item: Title: Study Design and Population Item: Title: Selection and Genotyping of Polymorphisms Item: Title: Statistical Analysis Item: Title: References Info: Title: VEGF Signaling Pathway Germline Polymorphisms as Prognostic Pharmacogenetic Biomarkers in Localized High-Grade Osteosarcoma Patients from the GEIS-33 Protocol Author: Juliana Salazar, María J. Arranz, Javier Martin-Broto, Massimo Serra, Emanuela Palmerini, Jeronimo Martínez-García, Antonio Juan-Ribelles, Maitane Andion, Javier Martinez-Trufero, Guiomar Gutiérrez, Aizpea Echebarria-Barona, Maria A. Vaz-Salgado, Alexandra Regueiro, Rosa Alvarez, Claudia Valverde, Pablo Luna, Pilar Blay and Ana Sebio Subject: Background/Objective: The vascular endothelial growth factor (VEGF) signaling pathway induces angiogenesis, which impacts tumor progression and clinical outcomes in patients with localized osteosarcoma. This study evaluates whether genetic polymorphisms in the VEGF signaling pathway are associated with survival outcomes in these patients. Methods: Sixty-nine patients with localized high-grade osteosarcoma enrolled in the GEIS-33 protocol and treated with MAP (methotrexate, doxorubicin, cisplatin) chemotherapy, surgery, and subsequent adjuvant treatment were included. Nine variants of interest in the VEGFA (rs1570360, rs2010963 and rs699947), FLT1 (VEGFR1; rs7993418, rs9513070 and rs9582036), and KDR (VEGFR2; rs1551641 and rs1870377 and rs2071559) genes were genotyped from peripheral blood samples using TaqMan Assay technology. Genetic data were correlated with relapse-free survival (RFS) and overall survival (OS) considering clinical variables as covariates. Results: The analyses showed nominally significant associations between the FLT1 variants rs7993418 and rs9582036 and survival. Patients carrying the rs7993418(C) allele had worse RFS (p = 0.01) and OS (p = 0.01). Carriers of the minor rs9582036(C) allele also had worse RFS (p = 0.02) and OS (p = 0.03). Additionally, patients harboring the TT genotype of the KDR rs1551641 variant had significantly worse RFS (p = 0.002). These polymorphisms remained statistically significant in the multivariate Cox regression analyses that included surgical margins and pathological response as covariates. Conclusions: Pharmacogenetics may contribute to precision medicine in oncology. Germline polymorphisms in the VEGF pathway may be useful as predictors of survival in high-grade localized osteosarcoma patients treated with chemotherapy, following validation in a large cohort of patients. Current treatment strategies aimed at improving outcomes for osteosarcoma patients may benefit from the identification of new biomarkers, such as these FLT1 rs7993418 and rs9582036 variants. 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