9f3713366611324452894923882016aa 296609.pdf a257c653888509f119e02f54ce3d206ce85c3470 296609.pdf 850a0c2ca53a52a7a3a1ce332eefdbed55fb13595b382b167d2adaac77ed38d6 296609.pdf Title: Exploring the Contribution of the Transporter AGT1/rBAT in Cystinuria Progression: Insights from Mouse Models and a Retrospective Cohort Study Subject: More than 20 years have passed since the identification of SLC3A1 and SLC7A9 as causative genes for cystinuria. However, cystinuria patients exhibit significant variability in the age of lithiasis onset, recurrence, and response to treatment, suggesting the presence of modulatory factors influencing cystinuria severity. In 2016, a second renal cystine transporter, AGT1, encoded by the SLC7A13 gene, was discovered. Although it was discarded as a causative gene for cystinuria, its possible effect as a modulatory gene remains unexplored. Thus, we analyzed its function in mouse models of cystinuria, screened the SLC7A13 gene in 34 patients with different lithiasic phenotypes, and functionally characterized the identified variants. Mice results showed that AGT1/rBAT may have a protective role against cystine lithiasis. In addition, among the four missense variants detected in patients, two exhibited a 25% impairment in AGT1/rBAT transport. However, no correlation between SLC7A13 genotypes and lithiasis phenotypes was observed in patients, probably because these variants were found in heterozygous states. In conclusion, our results, consistent with a previous study, suggest that AGT1/rBAT does not have a relevant effect on cystinuria patients, although an impact in patients carrying homozygous pathogenic variants cannot be discarded. Keywords: cystinuria; cystine transporter; rare disease; SLC7A13 variants; functional analysis Author: Clara Mayayo-Vallverdú, Esther Prat, Marta Vecino-Pérez, Laura González, Silvia Gràcia-Garcia, Luz San Miguel, Noelia Lopera, Angela Arias, Rafael Artuch, Miguel López de Heredia, Carlos Torrecilla, Ferran Rousaud-Barón, Oriol Angerri, Ekaitz Errasti-Murugarren and Virginia Nunes Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Dec 5 09:57:50 2023 CET ModDate: Tue Dec 5 10:05:27 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2117896 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- VVTBMF+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 XERKWR+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 UNFNAD+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 CRENWH+VnURWPalladioL Type 1 Custom yes yes no 23 0 IUYWSY+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 46 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 49 0 UBZRXD+CMSY10 Type 1 Builtin yes yes no 71 0 FJGLJA+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 81 0 FJGLJB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 87 0 FJGLJC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 93 0 FJGLKD+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 99 0 FJGLKE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 102 0 FJGLKF+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 105 0 FJGMLP+PalatinoLinotype-Roman TrueType MacRoman yes yes no 108 0 PYPUMW+URWPalladioL-BoldItal Type 1 Custom yes yes no 115 0 FJGLJA+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 125 0 FJGLJB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 131 0 FJGLJC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 137 0 FJGLKD+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 143 0 FJGLKE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 146 0 FJGLKF+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 149 0 FJGMLP+PalatinoLinotype-Roman TrueType MacRoman yes yes no 152 0 FJGLJA+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 163 0 FJGLJB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 169 0 FJGLJC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 175 0 FJGLKD+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 181 0 FJGLKE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 184 0 FJGLKF+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 187 0 FJGMLP+PalatinoLinotype-Roman TrueType MacRoman yes yes no 190 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 203 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-07-18 02:36:58 CEST RepresentationInformation: 296609.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-07-17 13:31:43 CEST Size: 2117896 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 371 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: AGT1/rBAT Contribution to Cystinuria Progression in Mouse Models Destination: subsection.2.1 Item: Title: SLC3A1, SLC7A9, and SLC7A13 Genetic Analysis in Cystinuria Patients Destination: subsection.2.2 Item: Title: Functional Studies of SLC7A13 Missense Variants Identified in Cystinuria Patients Destination: subsection.2.3 Item: Title: c.745G>A and c.988C>T SLC7A13 Variants Contribution to Amino Acid Excretion in Cystinuria Patients Destination: subsection.2.4 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Mouse Proceedings Destination: subsection.4.1 Item: Title: Brush-Border Western Blot Destination: subsection.4.2 Item: Title: Patients Destination: subsection.4.3 Item: Title: Patient Genotyping Destination: subsection.4.4 Item: Title: DNA Constructions and Site-Directed Mutagenesis Destination: subsection.4.5 Item: Title: Cell Culture and Transfection Destination: subsection.4.6 Item: Title: Visualization of GFP-Tagged Amino Acid Transporters Using Fluorescence Microscopy Destination: subsection.4.7 Item: Title: Amino Acid Transport Assay Destination: subsection.4.8 Item: Title: Statistical Analysis Destination: subsection.4.9 Item: Title: References Destination: section.A. Info: Title: Exploring the Contribution of the Transporter AGT1/rBAT in Cystinuria Progression: Insights from Mouse Models and a Retrospective Cohort Study Author: Clara Mayayo-Vallverdú, Esther Prat, Marta Vecino-Pérez, Laura González, Silvia Gràcia-Garcia, Luz San Miguel, Noelia Lopera, Angela Arias, Rafael Artuch, Miguel López de Heredia, Carlos Torrecilla, Ferran Rousaud-Barón, Oriol Angerri, Ekaitz Errasti-Murugarren and Virginia Nunes Subject: More than 20 years have passed since the identification of SLC3A1 and SLC7A9 as causative genes for cystinuria. However, cystinuria patients exhibit significant variability in the age of lithiasis onset, recurrence, and response to treatment, suggesting the presence of modulatory factors influencing cystinuria severity. In 2016, a second renal cystine transporter, AGT1, encoded by the SLC7A13 gene, was discovered. Although it was discarded as a causative gene for cystinuria, its possible effect as a modulatory gene remains unexplored. Thus, we analyzed its function in mouse models of cystinuria, screened the SLC7A13 gene in 34 patients with different lithiasic phenotypes, and functionally characterized the identified variants. Mice results showed that AGT1/rBAT may have a protective role against cystine lithiasis. In addition, among the four missense variants detected in patients, two exhibited a 25% impairment in AGT1/rBAT transport. However, no correlation between SLC7A13 genotypes and lithiasis phenotypes was observed in patients, probably because these variants were found in heterozygous states. In conclusion, our results, consistent with a previous study, suggest that AGT1/rBAT does not have a relevant effect on cystinuria patients, although an impact in patients carrying homozygous pathogenic variants cannot be discarded. 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