3009e71c0f9606dad9f7e2f545bff6e0 283225.pdf a89c962ddf9833636480a0bc11a51591c8fa731f 283225.pdf 6fa53e153754ec8a0e480f6907c906097959b09efb4b47549b26551027294fc3 283225.pdf Title: A Novel Intragenic Duplication in the HDAC8 Gene Underlying a Case of Cornelia de Lange Syndrome Subject: Cornelia de Lange syndrome (CdLS) is a multisystemic genetic disorder characterized by distinctive facial features, growth retardation, and intellectual disability, as well as various systemic conditions. It is caused by genetic variants in genes related to the cohesin complex. Single-nucleotide variations are the best-known genetic cause of CdLS; however, copy number variants (CNVs) clearly underlie a substantial proportion of cases of the syndrome. The NIPBL gene was thought to be the locus within which clinically relevant CNVs contributed to CdLS. However, in the last few years, pathogenic CNVs have been identified in other genes such as HDAC8, RAD21, and SMC1A. Here, we studied an affected girl presenting with a classic CdLS phenotype heterozygous for a de novo ~32 kbp intragenic duplication affecting exon 10 of HDAC8. Molecular analyses revealed an alteration in the physiological splicing that included a 96 bp insertion between exons 9 and 10 of the main transcript of HDAC8. The aberrant transcript was predicted to generate a truncated protein whose accessibility to the active center was restricted, showing reduced ease of substrate entry into the mutated enzyme. Lastly, we conclude that the duplication is responsible for the patient’s phenotype, highlighting the contribution of CNVs as a molecular cause underlying CdLS. Keywords: Cornelia de Lange syndrome; genetic disorder; copy number variants; HDAC8; intragenic duplication; array CGH; genetic diagnosis Author: Cristina Lucia-Campos, Irene Valenzuela, Ana Latorre-Pellicer, David Ros-Pardo, Marta Gil-Salvador, María Arnedo, Beatriz Puisac, Neus Castells, Alberto Plaja, Anna Tenes, Ivon Cuscó, Laura Trujillano, Feliciano J. Ramos, Eduardo F. Tizzano, Paulino Gómez-Puertas and Juan Pié Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Aug 10 09:28:56 2022 CEST ModDate: Mon Aug 22 12:21:48 2022 CEST 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: 1448603 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AHKDHL+VnURWPalladioL Type 1 Custom yes yes no 17 0 YEJUFV+URWPalladioL-Bold Type 1 Custom yes yes no 22 0 LWBWVH+URWPalladioL-Roma Type 1 Custom yes yes no 27 0 HGMICT+URWPalladioL-Ital Type 1 Custom yes yes no 31 0 YYTNHR+URWPalladioL-BoldItal Type 1 Custom yes yes no 35 0 XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 39 0 KONDUX+EuphemiaUCAS-Bold Type 1C WinAnsi yes yes no 55 0 HIZBRM+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 58 0 DYVFWR+CMSY10 Type 1 Builtin yes yes no 81 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 88 0 QDCWHF+EURM10 Type 1 Builtin yes yes no 92 0 NNOPJO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 106 0 NNOPJP+PalatinoLinotype CID TrueType Identity-H yes yes yes 109 0 NNOPKA+PalatinoLinotype TrueType WinAnsi yes yes no 115 0 NNOPKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 118 0 NNOPJO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 128 0 NNOPJP+PalatinoLinotype CID TrueType Identity-H yes yes yes 129 0 NNOPKA+PalatinoLinotype TrueType WinAnsi yes yes no 131 0 NNOPKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 132 0 NNOPJO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 143 0 NNOPJP+PalatinoLinotype CID TrueType Identity-H yes yes yes 144 0 NNOPKA+PalatinoLinotype TrueType WinAnsi yes yes no 146 0 NNOPKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 147 0 NNPDFO+TimesNewRoman CID TrueType Identity-H yes yes yes 148 0 NNOPJO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 161 0 NNOPJP+PalatinoLinotype CID TrueType Identity-H yes yes yes 162 0 NNOPKA+PalatinoLinotype TrueType WinAnsi yes yes no 164 0 NNOPKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 165 0 NNPDFO+TimesNewRoman CID TrueType Identity-H yes yes yes 166 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-11-08 02:29:54 CET RepresentationInformation: 283225.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-11-07 13:51:17 CET Size: 1448603 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 334 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: Clinical Diagnosis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Isolation of DNA and RNA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Next-Generation Sequencing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Array Comparative Genomic Hybridization Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: cDNA Synthesis and Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Real-Time Quantitative PCR (qPCR) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Structure Modeling of HDAC8 Variant and Molecular Dynamics Simulation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Children: Item: Title: Clinical Report Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: DNA Molecular Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: RNA Molecular Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: Structural Prediction of HDAC8 Variant Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@462d5aee Info: Title: A Novel Intragenic Duplication in the HDAC8 Gene Underlying a Case of Cornelia de Lange Syndrome Author: Cristina Lucia-Campos, Irene Valenzuela, Ana Latorre-Pellicer, David Ros-Pardo, Marta Gil-Salvador, María Arnedo, Beatriz Puisac, Neus Castells, Alberto Plaja, Anna Tenes, Ivon Cuscó, Laura Trujillano, Feliciano J. Ramos, Eduardo F. Tizzano, Paulino Gómez-Puertas and Juan Pié Subject: Cornelia de Lange syndrome (CdLS) is a multisystemic genetic disorder characterized by distinctive facial features, growth retardation, and intellectual disability, as well as various systemic conditions. It is caused by genetic variants in genes related to the cohesin complex. Single-nucleotide variations are the best-known genetic cause of CdLS; however, copy number variants (CNVs) clearly underlie a substantial proportion of cases of the syndrome. The NIPBL gene was thought to be the locus within which clinically relevant CNVs contributed to CdLS. However, in the last few years, pathogenic CNVs have been identified in other genes such as HDAC8, RAD21, and SMC1A. Here, we studied an affected girl presenting with a classic CdLS phenotype heterozygous for a de novo ~32 kbp intragenic duplication affecting exon 10 of HDAC8. Molecular analyses revealed an alteration in the physiological splicing that included a 96 bp insertion between exons 9 and 10 of the main transcript of HDAC8. The aberrant transcript was predicted to generate a truncated protein whose accessibility to the active center was restricted, showing reduced ease of substrate entry into the mutated enzyme. Lastly, we conclude that the duplication is responsible for the patient’s phenotype, highlighting the contribution of CNVs as a molecular cause underlying CdLS. 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