| Project/Area Number |
16390293
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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 |
Pediatrics
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| Research Institution | University of Tsukuba |
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Principal Investigator |
OTSU Makoto University of Tsukuba, Graduate Scool of Comprehensive Human Sciences, Assistant Professor, 大学院・人間総合科学研究科, 講師 (30361330)
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| Co-Investigator(Kenkyū-buntansha) |
ARIGA Tadashi Hokkaido University, Dept.of Pediatrics, Professor, 大学院・医学研究科, 教授 (60322806)
MIYAMOTO Masaki Hokkaido University, Hokkaido Univ.Hospital, Assistant Professor, 北海道大学病院, 助手 (40333611)
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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 |
¥14,500,000 (Direct Cost: ¥14,500,000)
Fiscal Year 2005: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2004: ¥12,000,000 (Direct Cost: ¥12,000,000)
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| Keywords | CXCR4 / CXCL12 / hematopietic stem cell transplantation / graft failure / retroviral vector / WHIM syndrome / 血液幹細胞 / 骨髄移植 / レトロウイルベクター / WHIM |
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| Research Abstract |
Objective : Studies have suggested that the stromal derived factor-1(SDF-1) and its receptor CXCR4 play an important role in HSC (hematopoietic stem cell) homing and engraftment upon transplantation. Recently, C-terminus truncation of CXCR4 has been identified as causative mutations in WHIM syndrome, an immune deficiency disorder characterized by Warts, Hypogammaglobulinemia, Infections and Myelokathexis. We have tested if forced expression of truncated CXCR4 (WHIM mutants) could confer enhanced responsiveness to SDF-1 on murine HSC using a retroviral vector system.
Methods : We constructed retroviral vectors that express wild type murine CXCR4 or WHIM mutants using the pGCsam-backbone, then generated highly concentrated retroviral particles pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G).
Erythroleukemia K562 cells were transduced with the above retroviral vectors to test functionality of the mutant CXCR4 molecules. We also transduced HSC-rich marrow populations to test responsiveness to SDF-1 of murine HSCs expressing WHIM mutants in both in vitro and in vivo experimental systems.
Results : Both K562 and murine c-Kit positive/lineage-marker negative BM cells were successfully transduced with all the retrovi Future directions : Establishment of the reliable in vivo homing assay must be achieved for future experiments.
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