c0adf1b78e1c5bd31fa6701812f1ea13 pmc_10671092.pdf 60e955e09ea53b5f2e82e4f325e1e90e5286e5ae pmc_10671092.pdf 03ada0a21302d76e66e5445f170ee95691d79b11f3cc2fb13c0db896406a3ece pmc_10671092.pdf Title: Key Genes of the Immune System and Predisposition to Acquired Hemophilia A: Evidence from a Spanish Cohort of 49 Patients Using Next-Generation Sequencing Subject: Acquired hemophilia A (AHA) is a rare bleeding disorder caused by the presence of autoantibodies against factor VIII (FVIII). As with other autoimmune diseases, its etiology is complex and its genetic basis is unknown. The aim of this study was to identify the immunogenetic background that predisposes individuals to AHA. HLA and KIR gene clusters, as well as KLRK1, were sequenced using next-generation sequencing in 49 AHA patients. Associations between candidate genes involved in innate and adaptive immune responses and AHA were addressed by comparing the alleles, genotypes, haplotypes, and gene frequencies in the AHA cohort with those in the donors’ samples or Spanish population cohort. Two genes of the HLA cluster, as well as rs1049174 in KLRK1, which tags the natural killer (NK) cytotoxic activity haplotype, were found to be linked to AHA. Specifically, A*03:01 (p = 0.024; odds ratio (OR) = 0.26[0.06–0.85]) and DRB1*13:03 (p = 6.8 103, OR = 7.56[1.64–51.40]), as well as rs1049174 (p = 0.012), were significantly associated with AHA. In addition, two AHA patients were found to carry one copy each of the low-frequency allele DQB1*03:09 (nallele = 2, 2.04%), which was completely absent in the donors. To the best of our knowledge, this is the first time that the involvement of these specific alleles in the predisposition to AHA has been proposed. Further molecular and functional studies will be needed to unravel their specific contributions. We believe our findings expand the current knowledge on the genetic factors involved in susceptibility to AHA, which will contribute to improving the diagnosis and prognosis of AHA patients. Keywords: rare bleeding disorders; Acquired hemophilia A; autoantibodies; next-generation sequencing; genetic predisposition to disease; immunogenetics Author: Jose Pardos-Gea, Laura Martin-Fernandez, Laia Closa, Ainara Ferrero, Cristina Marzo, Manuel Rubio-Rivas, Francesca Mitjavila, José Ramón González-Porras, José María Bastida, José Mateo, Marina Carrasco, Ángel Bernardo, Itziar Astigarraga, Reyes Aguinaco, Irene Corrales, Iris Garcia-Martínez and Francisco Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Nov 15 14:14:10 2023 CET ModDate: Wed Nov 15 14:31:03 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 658770 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- FFAYKL+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 XOBIIF+VnURWPalladioL Type 1 Custom yes yes no 15 0 WMFALP+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 TXUJEB+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 JIRAVE+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 RZOAPT+CMSY10 Type 1 Builtin yes yes no 32 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 50 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 53 0 MAKFHF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 88 0 MAKFHG+PalatinoLinotype CID TrueType Identity-H yes yes yes 91 0 MAKFHH+PalatinoLinotype TrueType WinAnsi yes yes no 97 0 MAKFHJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 100 0 MAKGGH+PalatinoLinotype TrueType MacRoman yes yes no 103 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 110 0 BOLMEN+SFRM0900 Type 1 Custom yes yes no 136 0 CKBPVA+EURM10 Type 1 Builtin yes yes no 144 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-03 02:40:43 CEST RepresentationInformation: pmc_10671092.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-24 02:38:49 CEST Size: 658770 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 383 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: Demographic and Clinical Profiles of the AHA Cohort Destination: subsection.2.1 Item: Title: Hemostatic Treatment and Immunosuppressive Therapy Destination: subsection.2.2 Item: Title: Genetic Analyses and Association Studies Destination: subsection.2.3 Children: Item: Title: Classical HLA Class I and II Genes Destination: subsubsection.2.3.1 Item: Title: Non-Classical HLA Class I and HLA Class I-like Genes Destination: subsubsection.2.3.2 Item: Title: KIR Genes, Genotypes, and Haplotypes Destination: subsubsection.2.3.3 Item: Title: KLRK1 Gene Destination: subsubsection.2.3.4 Item: Title: Discussion Destination: section.3 Item: Title: Methods Destination: section.4 Children: Item: Title: Patients Destination: subsection.4.1 Item: Title: Sample Collection and Phenotype Determinations Destination: subsection.4.2 Item: Title: Candidate Region Amplification and Sequencing Destination: subsection.4.3 Item: Title: Bioinformatics Analyses Destination: subsection.4.4 Item: Title: Reference Samples Destination: subsection.4.5 Item: Title: Statistical Analyses Destination: subsection.4.6 Item: Title: References Destination: section.A. Info: Title: Key Genes of the Immune System and Predisposition to Acquired Hemophilia A: Evidence from a Spanish Cohort of 49 Patients Using Next-Generation Sequencing Author: Jose Pardos-Gea, Laura Martin-Fernandez, Laia Closa, Ainara Ferrero, Cristina Marzo, Manuel Rubio-Rivas, Francesca Mitjavila, José Ramón González-Porras, José María Bastida, José Mateo, Marina Carrasco, Ángel Bernardo, Itziar Astigarraga, Reyes Aguinaco, Irene Corrales, Iris Garcia-Martínez and Francisco Vidal Subject: Acquired hemophilia A (AHA) is a rare bleeding disorder caused by the presence of autoantibodies against factor VIII (FVIII). As with other autoimmune diseases, its etiology is complex and its genetic basis is unknown. The aim of this study was to identify the immunogenetic background that predisposes individuals to AHA. HLA and KIR gene clusters, as well as KLRK1, were sequenced using next-generation sequencing in 49 AHA patients. Associations between candidate genes involved in innate and adaptive immune responses and AHA were addressed by comparing the alleles, genotypes, haplotypes, and gene frequencies in the AHA cohort with those in the donors’ samples or Spanish population cohort. Two genes of the HLA cluster, as well as rs1049174 in KLRK1, which tags the natural killer (NK) cytotoxic activity haplotype, were found to be linked to AHA. Specifically, A*03:01 (p = 0.024; odds ratio (OR) = 0.26[0.06 0.85]) and DRB1*13:03 (p = 6.8 103, OR = 7.56[1.64 51.40]), as well as rs1049174 (p = 0.012), were significantly associated with AHA. In addition, two AHA patients were found to carry one copy each of the low-frequency allele DQB1*03:09 (nallele = 2, 2.04%), which was completely absent in the donors. To the best of our knowledge, this is the first time that the involvement of these specific alleles in the predisposition to AHA has been proposed. Further molecular and functional studies will be needed to unravel their specific contributions. We believe our findings expand the current knowledge on the genetic factors involved in susceptibility to AHA, which will contribute to improving the diagnosis and prognosis of AHA patients. 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