PMID- 36968585 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20230328 IS - 1664-8021 (Print) IS - 1664-8021 (Electronic) IS - 1664-8021 (Linking) VI - 14 DP - 2023 TI - cDNA sequencing increases the molecular diagnostic yield in Chediak-Higashi syndrome. PG - 1072784 LID - 10.3389/fgene.2023.1072784 [doi] LID - 1072784 AB - Introduction: Chediak-Higashi syndrome (CHS) is rare autosomal recessive disorder caused by bi-allelic variants in the Lysosomal Trafficking Regulator (LYST) gene. Diagnosis is established by the detection of pathogenic variants in LYST in combination with clinical evidence of disease. Conventional molecular genetic testing of LYST by genomic DNA (gDNA) Sanger sequencing detects the majority of pathogenic variants, but some remain undetected for several individuals clinically diagnosed with CHS. In this study, cDNA Sanger sequencing was pursued as a complementary method to identify variant alleles that are undetected by gDNA Sanger sequencing and to increase molecular diagnostic yield. Methods: Six unrelated individuals with CHS were clinically evaluated and included in this study. gDNA Sanger sequencing and cDNA Sanger sequencing were performed to identify pathogenic LYST variants. Results: Ten novel LYST alleles were identified, including eight nonsense or frameshift variants and two in-frame deletions. Six of these were identified by conventional gDNA Sanger sequencing; cDNA Sanger sequencing was required to identify the remaining variant alleles. Conclusion: By utilizing cDNA sequencing as a complementary technique to identify LYST variants, a complete molecular diagnosis was obtained for all six CHS patients. In this small CHS cohort, the molecular diagnostic yield was increased, and canonical splice site variants identified from gDNA Sanger sequencing were validated by cDNA sequencing. The identification of novel LYST alleles will aid in diagnosing patients and these molecular diagnoses will also lead to genetic counseling, access to services and treatments and clinical trials in the future. CI - Copyright (c) 2023 Kuptanon, Morimoto, Nicoli, Stephen, Yarnell, Dorward, Owen, Parikh, Ozbek, Malbora, Ciccone, Gunay-Aygun, Gahl, Introne and Malicdan. FAU - Kuptanon, Chulaluk AU - Kuptanon C AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Morimoto, Marie AU - Morimoto M AD - National Institutes of Health Undiagnosed Diseases Program, National Institutes of Health Common Fund, Office of the Director, National Institutes of Health, Bethesda, MD, United States. FAU - Nicoli, Elena-Raluca AU - Nicoli ER AD - National Institutes of Health Undiagnosed Diseases Program, National Institutes of Health Common Fund, Office of the Director, National Institutes of Health, Bethesda, MD, United States. FAU - Stephen, Joshi AU - Stephen J AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Yarnell, David S AU - Yarnell DS AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Dorward, Heidi AU - Dorward H AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Owen, William AU - Owen W AD - Children's Hospital of The King's Daughters, Norfolk, VA, United States. FAU - Parikh, Suhag AU - Parikh S AD - Department of Pediatrics, School of Medicine, Emory University, Atlanta, GA, United States. FAU - Ozbek, Namik Yasar AU - Ozbek NY AD - Division of Pediatric Hematology and Oncology, University of Yeni Yuzyil, Gaziosmanpasa Hospital, Istanbul, Turkiye. FAU - Malbora, Baris AU - Malbora B AD - Department of Pediatric Hematology/Oncology, Ankara City Hospital, The University of Health Sciences, Ankara, Turkiye. FAU - Ciccone, Carla AU - Ciccone C AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Gunay-Aygun, Meral AU - Gunay-Aygun M AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Gahl, William A AU - Gahl WA AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. AD - National Institutes of Health Undiagnosed Diseases Program, National Institutes of Health Common Fund, Office of the Director, National Institutes of Health, Bethesda, MD, United States. FAU - Introne, Wendy J AU - Introne WJ AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. AD - Office of the Clinical Director, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. FAU - Malicdan, May Christine V AU - Malicdan MCV AD - Human Biochemical Genetics Section, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, United States. AD - National Institutes of Health Undiagnosed Diseases Program, National Institutes of Health Common Fund, Office of the Director, National Institutes of Health, Bethesda, MD, United States. LA - eng PT - Journal Article DEP - 20230308 PL - Switzerland TA - Front Genet JT - Frontiers in genetics JID - 101560621 PMC - PMC10031035 OTO - NOTNLM OT - LYST OT - molecular diagnosis OT - monogenic diseases OT - oculocutaneous albinism OT - personalized medicine OT - rare disorders OT - splicing abnormalities COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2023/03/28 06:00 MHDA- 2023/03/28 06:01 PMCR- 2023/03/08 CRDT- 2023/03/27 03:27 PHST- 2022/10/17 00:00 [received] PHST- 2023/02/22 00:00 [accepted] PHST- 2023/03/27 03:27 [entrez] PHST- 2023/03/28 06:00 [pubmed] PHST- 2023/03/28 06:01 [medline] PHST- 2023/03/08 00:00 [pmc-release] AID - 1072784 [pii] AID - 10.3389/fgene.2023.1072784 [doi] PST - epublish SO - Front Genet. 2023 Mar 8;14:1072784. doi: 10.3389/fgene.2023.1072784. eCollection 2023.