Sensitive detection of GATA1 mutations using complementary DNA-based analysis for transient abnormal myelopoiesis associated with the Down syndrome
Corresponding Author
Shumpei Mizuta
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, Hyogo, Japan
Correspondence
Shumpei Mizuta, P Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, 2-17-77 Higashinaniwa-cho Amagasaki, Hyogo, Japan.
Email: [email protected]
Search for more papers by this authorNoriko Yamane
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorSaya Mononobe
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorAsami Watanabe
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorRitsuko Kitamura
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTadamori Takahara
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorChieko Matsushima
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorAtushi Yoshida
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorSeiji Okamoto
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorKuniaki Tanaka
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Search for more papers by this authorAtsushi Iwai
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Search for more papers by this authorAtsuko Ikegawa
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTakahito Wada
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Genomic Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorIkuya Usami
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorToshiro Maihara
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTakao Komai
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorToshio Heike
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorYoshinobu Nishida
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorKenichiro Kobayashi
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Department of Pediatric Hematology and Oncology Research, Research Institute, National Center for Child Health and Development, Tokyo, Japan
Search for more papers by this authorCorresponding Author
Shumpei Mizuta
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, Hyogo, Japan
Correspondence
Shumpei Mizuta, P Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, 2-17-77 Higashinaniwa-cho Amagasaki, Hyogo, Japan.
Email: [email protected]
Search for more papers by this authorNoriko Yamane
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorSaya Mononobe
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorAsami Watanabe
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorRitsuko Kitamura
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTadamori Takahara
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorChieko Matsushima
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorAtushi Yoshida
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorSeiji Okamoto
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorKuniaki Tanaka
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Search for more papers by this authorAtsushi Iwai
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Search for more papers by this authorAtsuko Ikegawa
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTakahito Wada
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Genomic Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorIkuya Usami
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorToshiro Maihara
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorTakao Komai
Department of Clinical Laboratory, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorToshio Heike
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorYoshinobu Nishida
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Neonatology, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Search for more papers by this authorKenichiro Kobayashi
Department of Pediatrics, Hyogo Prefectural Amagasaki General Medical Center, Amagasaki, Japan
Department of Pediatric Hematology and Oncology, Hyogo Prefectural Amagasaki General Medical Center, Hyogo, Japan
Department of Pediatric Hematology and Oncology Research, Research Institute, National Center for Child Health and Development, Tokyo, Japan
Search for more papers by this authorFunding information
This work was supported by JSPS KAKENHI Grant Number JP18K15390
Abstract
Introduction
GATA1 mutation plays an important role in initiating transient abnormal myelopoiesis (TAM) and in the clonal evolution towards acute megakaryoblastic leukaemia (AMKL) associated with Down syndrome (DS). This study aimed to develop and validate the clinical utility of a complementary DNA (cDNA) analysis in parallel with the conventional genomic DNA (gDNA) Sanger sequencing (Ss), as an initial screening test for GATA1 mutations.
Methods
GATA1 mutations were evaluated using both gDNA and cDNA in 14 DS patients using Ss and fragment analysis (FA), respectively.
Results
The detection sensitivity of conventional gDNA sequencing was limited in low blast percentage TAM (LBP-TAM); however, cDNA-based Ss readily detected all the pathognomonic GATA1 mutations. The cDNA-based FA readily detected GATA1 frameshift mutation with a reliable sensitivity ranging from 0.005% to 0.01% of clonal cells.
Conclusions
GATA1 mutations are heterogeneous; therefore, we would like to propose a dual cDNA and gDNA analysis as a standard diagnostic approach, especially for LBP-TAM. cDNA-based FA promises an excellent sensitivity for detecting frameshift GATA1 mutations in the longitudinal clonal evolution towards AMKL without using a patient specific primer.
CONFLICT OF INTEREST
The authors have no competing interests.
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