Elucidation of molecular basis of May-Hegglin anomaly and its related disordes
| Project/Area Number |
16390283
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | National Hospital Organization Nagoya Medical Center, Clinical Research Center |
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Principal Investigator |
SAITO Hidehiko National Hospital Organization Nagoya Medical Center, President, 名古屋医療センター, 院長 (20153819)
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| Co-Investigator(Kenkyū-buntansha) |
HAMAGUCHI Motohiro National Hospital Organization Nagoya Medical Center, Clinical Research Center, Head, 臨床研究センター・止血血栓研究部, 部長 (30393177)
YAMAZAKI Tomio National Hospital Organization Nagoya Medical Center, Clinical Research Center, Chief, 臨床研究センター・止血血栓研究部, 室長 (00282202)
KUNISHIMA Shinji National Hospital Organization Nagoya Medical Center, Clinical Research Center, Chief, 臨床研究センター・止血血栓研究部, 室長 (60373495)
KOJIMA Tetsuhito Nagoya University, Department of Medicine, Professor, 医学部, 教授 (40161913)
MATSUSHITA Tadashi Nagoya University Hospital, Department of Hematology, Assistant Professor, 医学部附属病院, 助手 (30314008)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2005: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2004: ¥9,600,000 (Direct Cost: ¥9,600,000)
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| Keywords | May-Hegglin anomaly / MYH9 disorders / NMMHCA / Congenital platelet disorders / Congenital thrombocytopenia / 血小板機能異常症 / 血小板減少症 |
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| Research Abstract |
Analysis of MYH9 disorders
We studied the neutrophil NMMHCA localization in 10 patients with MYH9 disorders. In five cases, leukocyte inclusion bodies were observed on May-Grunwald-Giemsa stained peripheral blood smears. In the rest five cases, the presence of leukocyte inclusion bodies were ambiguous. Abnormal staining of neutrophil NMMHCA was detected in all cases. Subsequent mutational analysis of the MYH9 gene showed that all cases had a heterozygous MYH9 mutation. Immunofluorescence analysis of neutrophil NMMHCA localization represents a clear and unambiguous alternative to conventional staining for the detection of minute leukocyte inclusions and the diagnosis of the autosomal dominant macrothrombocytopenias caused by MYH9 mutations.
Establishment of MYH9 Knock in mice
Mouse genomic DNA clones were isolated from the 129SvJ-derived genomic library, and the clones spanning the exon 16 region were used to construct the targeting vector. A R702C point mutation was introduced by site-directed mutagenesis. At the 3' end, a LoxP-Neo-LoxP and DTA cassettes were inserted for positive and negative selection of electroporated ES cells, respectively. The targeting vector was electroporated to ES cells, and the homologous recombinants were selected by PCR and Southern blotting. The knock-in mice are under construction by crossing chimeric founders. After establishment of MYH9 R702C knock in mice, detailed pathohistological and physiological examinations of blood cells, kidney and inner ear will be investigated.
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Report
(3 results)
Research Products
(10 results)
