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2757 Diagnostic Whole Exome Sequencing for 166 Patients with Inherited Bone Marrow Failure Syndrome

Program: Oral and Poster Abstracts
Session: 508. Bone Marrow Failure: Poster III
Hematology Disease Topics & Pathways:
Adult, Diseases, Bone Marrow Failure, Pediatric, Technology and Procedures, Study Population, Clinically relevant, NGS
Monday, December 7, 2020, 7:00 AM-3:30 PM

Motoharu Hamada1*, Hideki Muramatsu, MD, PhD2, Yusuke Okuno, MD, PhD3, Ayako Yamamori, MD4*, Taro Yoshida, MD4*, Masayuki Imaya, MD4*, Manabu Wakamatsu, MD1*, Shunsuke Miwata, MD1*, Kotaro Narita, MD, PhD1*, Daisuke Ichikawa, MD1*, Rieko Taniguchi, MD, PhD1*, Atsushi Narita, MD, PhD5*, Nozomu Kawashima, MD, PhD1*, Eri Nishikawa, MD, PhD1*, Nobuhiro Nishio, MD, PhD1,6*, Asahito Hama, MD, PhD7*, Masafumi Ito, MD, PhD8*, Tomoo Ogi, Ph.D.9*, Seiji Kojima, MD, PhD1* and Yoshiyuki Takahashi, MD, PhD1*

1Department of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan
2Nagoya University Graduate School of Medicine, Nagoya, Japan
3Medical Genomics Center, Nagoya University Hospital, Nagoya, Japan
4Pediatrics, Department of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan
5Department of Pediatrics, Graduate School of Medicine, Nagoya University, Nagoya, Japan
6Center for Advanced Medicine and Clinical Research, Nagoya University Hospital, Nagoya, Japan
7Department of Hematology and Oncology, Children's Medical Center, Japanese Red Cross Nagoya First Hospital, Nagoya, Japan
8Department of Pathology, Japanese Red Cross Nagoya First Hospital, Nagoya, Japan
9Department of Genetics, Research Institute of Environmental Medicine (RIeM), Nagoya University, Nagoya, Japan

BACKGROUND: Inherited bone marrow failure syndromes (IBMFSs) are a heterogeneous group of genetic disorders characterized by bone marrow failure, physical anomalies, and various kinds of organ complications. In addition to classical IBMFSs, such as Fanconi anemia, Diamond-Blackfan anemia, Dyskeratosis congenita, Shwachman-Diamond syndrome, and familial platelet disorders, many types of unclassified IBMFSs are reported. Over 100 genes are considered causative genes; however, the precise genetic diagnosis of IBMFSs remains challenging. We developed a capture-based target sequencing method for IBMFSs that covers more than 180 associated genes. Our system achieved genetic diagnosis for 225 (35%) of 738 patients between 2013 and 2018. However, the causative gene remained unknown for 513 (65%) patients, and further genetic analysis of these “target-negative” cases was necessary to achieve a precise diagnosis.

METHODS: We performed whole exome sequencing (WES) for patients who were “target-negative” but strongly suspected of having IBMFS based on the following clinical characteristics: physical or organ anomalies (skin, nail, hair, skeletal, growth, cardiac, lung, liver, or genitourinary), family history of hematological disorder, young age (≤2 years), short telomere length (<-2.0 SD), and hyper sensitivity to the chromosome breakage test. A sequencing library was prepared using the SureSelect Human All Exon 50Mb kit (Agilent Technologies, Santa Clara, CA, USA) and it was sequenced using the HiSeq2000 platform (Illumina, San Diego, CA, USA), according to manufacturers’ instructions. The candidate germline variants were detected through our Genomon-exome analysis pipeline. With mean coverage of 100×, ≥ 85% of all protein coding bases were covered at 20× or more.

RESULTS: Among the 513 “target-negative” cases, 166 patients were evaluated, of whom 17 patients’ parents were also analyzed in a trio-based analysis. New pathogenic variants were identified in 18 of the 166 (11%) patients according to the American College of Medical Genetics (ACMG) guidelines, of which 5 variants were revealed to be de novo. Diagnostic variants were identified in FANCF, SRP54, RPL19, RPL5, RTEL1, RUNX1, MECOM, CDC42, GNE, SLNF14 (all n = 1). In addition to IBMFS-associated genes, causative genes for congenital hemolytic anemia (G6PD, PKLR), inborn error of metabolism (SLC46A1), and primary immune deficiency (NFKB2, LRBA) are also identified (all n = 1). Moreover, loss-of-function mutation of ADH5 gene are identified in three patients that seems to be associated to novel IBMFSs. On the other hand, no pathogenic variant in GATA2, ERCC6L2, LIG4, and SAMD9/SAMD9L genes that are reported as unclassified IBMFSs in Europe and United States are identified in our cohort.

CONCLUSION: Our findings support the utility of WES (especially trio-based analysis) as a diagnostic tool for IBMFSs. Furthermore, genetic background of IBMFSs in East Asia seems to be different from that of Europe and United States.

Disclosures: No relevant conflicts of interest to declare.

*signifies non-member of ASH