2016 Fiscal Year Research-status Report
Morphometrical and quantitative transcriptome analyses of C-C chemokine receptor 5 in functional structure of osteoclasts
| Project/Area Number |
16K20412
|
|---|
| Research Institution | Ehime University |
|---|
Principal Investigator |
李 智媛 愛媛大学, プロテオサイエンスセンター, 助教(特定教員) (70711274)
|
|---|
| Project Period (FY) |
2016-04-01 – 2018-03-31
|
| Keywords | chemokine receptor 5 / osteoclast / bone / RNA-sequence / RANKL |
|---|
| Outline of Annual Research Achievements |
[Purpose of research] Our previous observations found that chemokine receptor 5 (CCR5) had unique role for osteoclastic functions. To further investigate this research results, we will take combinatorial approaches of morphometericla, cytomorphhometrical and transcriptome analyses. The putative results would patio-temporally dissociate chemokine-mediated signals during osteoclast ontogeny and possibly contribute to therapeutic development for osteoporosis. In 1st year of this project, we conducted half of project as planned described above.
[Archievements] 1) Histomorphometrial analysis was conducted on femoral bone sections obtained from 8-10 week-old Ccr5 wild type and Ccr5 knock out mice; Ccr5-deficient mice had significantly increased number and size of osteoclasts, although they did not show significant difference in BMD compared to their wild-type littermates. In keeping with osteoclst dysfunction, Ccr5-deficient mice were less susceptible to RANKL-induced bone loss. 2) Transcriptome analysis was confirmed from chemokine knockout bone marrow cell culture, and Data of RNA-seq were analyzed using specific software; Maldi-Tof and RNA-sequencing demonstrated that CCR5-mediated signaling, cooperating with RANKL-mediated signaling, regulated cellular architecture and motility of differentiated osteoclast through activation of small-GTPases.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The first year of project is being under processing as planed.
|
| Strategy for Future Research Activity |
1) Cytomorphometrical analysis of Ccr5+/+ and Ccr5-/- osteoclasts by fluorescence bio-imaging; To evaluate three-dimensional architecture of cultured osteoclasts from Ccr5+/+ and Ccr5-/-, we will take 3D fluorescence images by confocal based super resolution microscopy.
2) Validation of candidate factor and signaling pathway from profiling; we will construct RNA interference (RANi) plasmid to block expression of candidate molecules using basic vector. These cells are analyzed by osteoclastic marker expression by qRT-PCR, osteoclast function assay and cytomorphometrical analyses.
|
| Causes of Carryover |
budget for next year experiment
|
| Expenditure Plan for Carryover Budget |
be expect to spend for data analysis of RNA sequencing
|
-
[Journal Article] Bone Research2017
Author(s)
Aya Tanaka, Ji-Won Lee, Kyoko Hirano, Yukihiro Isogai, Toshinori Ishizuya, Rhoko Takao-Kawabata, Tadahiro Iimura
-
Journal Title
Administration frequency as well as dosage of PTH are associated with development of cortical porosity in ovariwctomized rats
Volume: 5
Pages: -
DOI
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
-
[Journal Article] PLOS One2017
Author(s)
Hiroshi Yamane, Aya Takakura, Yukari Shimadzu, Toshiyuki Kodama, Ji-Won Lee, Yukihiro Isogai, Toshinori Ishizuya Rhoko Takao-Kawabata, Tadahiro Iimura
-
Journal Title
Acute development of cortical porosity and endosteal naive bone formation from the daily but not weekly short-term administration of PTH in rabbit
Volume: -
Pages: -
DOI
Peer Reviewed / Open Access / Int'l Joint Research
-
[Journal Article] Journal of Oral Biosciences2017
Author(s)
Tadahiro Iimura, Ji-Won Lee
-
Journal Title
Unveiling a rhythmic regulatory mode hidden in developmental tissue growth by fluorescence live imaging-based mathematical modeling
Volume: 59
Pages: 6-11
DOI
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
-
[Journal Article] Japanse Dental Science Review2017
Author(s)
Ji-Won Lee, Tadahiro Iimura
-
Journal Title
Shedding quantitative fluorescence light on novel regulatory mechanisms in skeletal biomedicine and biodentistry
Volume: 53
Pages: 2-10
DOI
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
-
-
-
-
-
-
-
-
[Presentation] C-C chemokine receptor 5, a co-receptor of HIV, -mediated signal is required for geometric architecture and function of osteoclasts, thus for RANKL-induced bone destruction2016
Author(s)
Ji-Won Lee, Akiyoshi Hoshino, Kazuki Inoue, Shunsuke Uehara, Yasuhiro Kobayashi, Satoshi Ueha, Kouji Matsushima, Akira Yamaguchi, Yuuki Imai, Tadahiro Iimura
Organizer
43rd Annual European Calcified Tissue Society Congress
Place of Presentation
Rome, Italy
Year and Date
2016-05-14 – 2016-05-17
Int'l Joint Research
