Abstract
Recently, autophagy-related proteins were shown to regulate osteoclast mediated bone resorption, a critical process in autoimmune diseases such as rheumatoid arthritis. However, the role of autophagy-linked FYVE containing protein, WDFY3, in osteoclast biology remains elusive. WDFY3 is a master regulator in selective autophagy for clearing ubiquitinated protein aggregates and has been linked with rheumatoid arthritis. Herein, we used a series of WDFY3 transgenic mice (Wdfy3 lacZ and Wdfy3 loxP ) to investigate the function of WDFY3 in osteoclast development and function. Our data demonstrate that WDFY3 is highly expressed at the growth plate of neonatal mice and is expressed in osteoclasts in vitro cultures. Osteoclasts derived from WDFY3 conditional knockout mice (Wdfy3 loxP/loxP -LysM-Cre + ) demonstrated increased osteoclast differentiation as evidenced by higher number and enlarged size of TRAP+ multinucleated cells. Western blot analysis also revealed up-regulation of TRAF6 and an increase in RANKL-induced NF-κB signaling in WDFY3-deficient bone marrow-derived macrophages compared to wild type cultures. Consistent with these observations WDFY3-deficient cells also demonstrated an increase in osteoclast-related genes Ctsk, Acp5, Mmp9 and an increase of dentine resorption in in vitro assays. Importantly, in vivo RANKL gene transfer exacerbated bone loss in WDFY3 conditional knockout mice, as evidenced by elevated serum TRAP, CTX-I and micro-CT analysis of distal femurs compared to wild type littermates. Taken together, our data highlight a novel role for WDFY3 in osteoclast development and function, which can be exploited for the treatment of musculoskeletal diseases.
| Original language | English (US) |
|---|---|
| Journal | Journal of Autoimmunity |
| DOIs | |
| State | Accepted/In press - May 12 2016 |
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Keywords
- Autophagy
- Autophagy-linked FYVE containing protein
- Musculoskeletal diseases
- Osteoclast
- TRAF6
- WDFY3
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Autophagy-linked FYVE containing protein WDFY3 interacts with TRAF6 and modulates RANKL-induced osteoclastogenesis. / Wu, Dennis J.; Gu, Ran; Sarin, Ritu; Zavodovskaya, Regina; Chen, Chia Pei; Christiansen, Blaine A; Adamopoulos, Iannis.
In: Journal of Autoimmunity, 12.05.2016.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Autophagy-linked FYVE containing protein WDFY3 interacts with TRAF6 and modulates RANKL-induced osteoclastogenesis
AU - Wu, Dennis J.
AU - Gu, Ran
AU - Sarin, Ritu
AU - Zavodovskaya, Regina
AU - Chen, Chia Pei
AU - Christiansen, Blaine A
AU - Adamopoulos, Iannis
PY - 2016/5/12
Y1 - 2016/5/12
N2 - Recently, autophagy-related proteins were shown to regulate osteoclast mediated bone resorption, a critical process in autoimmune diseases such as rheumatoid arthritis. However, the role of autophagy-linked FYVE containing protein, WDFY3, in osteoclast biology remains elusive. WDFY3 is a master regulator in selective autophagy for clearing ubiquitinated protein aggregates and has been linked with rheumatoid arthritis. Herein, we used a series of WDFY3 transgenic mice (Wdfy3 lacZ and Wdfy3 loxP ) to investigate the function of WDFY3 in osteoclast development and function. Our data demonstrate that WDFY3 is highly expressed at the growth plate of neonatal mice and is expressed in osteoclasts in vitro cultures. Osteoclasts derived from WDFY3 conditional knockout mice (Wdfy3 loxP/loxP -LysM-Cre + ) demonstrated increased osteoclast differentiation as evidenced by higher number and enlarged size of TRAP+ multinucleated cells. Western blot analysis also revealed up-regulation of TRAF6 and an increase in RANKL-induced NF-κB signaling in WDFY3-deficient bone marrow-derived macrophages compared to wild type cultures. Consistent with these observations WDFY3-deficient cells also demonstrated an increase in osteoclast-related genes Ctsk, Acp5, Mmp9 and an increase of dentine resorption in in vitro assays. Importantly, in vivo RANKL gene transfer exacerbated bone loss in WDFY3 conditional knockout mice, as evidenced by elevated serum TRAP, CTX-I and micro-CT analysis of distal femurs compared to wild type littermates. Taken together, our data highlight a novel role for WDFY3 in osteoclast development and function, which can be exploited for the treatment of musculoskeletal diseases.
AB - Recently, autophagy-related proteins were shown to regulate osteoclast mediated bone resorption, a critical process in autoimmune diseases such as rheumatoid arthritis. However, the role of autophagy-linked FYVE containing protein, WDFY3, in osteoclast biology remains elusive. WDFY3 is a master regulator in selective autophagy for clearing ubiquitinated protein aggregates and has been linked with rheumatoid arthritis. Herein, we used a series of WDFY3 transgenic mice (Wdfy3 lacZ and Wdfy3 loxP ) to investigate the function of WDFY3 in osteoclast development and function. Our data demonstrate that WDFY3 is highly expressed at the growth plate of neonatal mice and is expressed in osteoclasts in vitro cultures. Osteoclasts derived from WDFY3 conditional knockout mice (Wdfy3 loxP/loxP -LysM-Cre + ) demonstrated increased osteoclast differentiation as evidenced by higher number and enlarged size of TRAP+ multinucleated cells. Western blot analysis also revealed up-regulation of TRAF6 and an increase in RANKL-induced NF-κB signaling in WDFY3-deficient bone marrow-derived macrophages compared to wild type cultures. Consistent with these observations WDFY3-deficient cells also demonstrated an increase in osteoclast-related genes Ctsk, Acp5, Mmp9 and an increase of dentine resorption in in vitro assays. Importantly, in vivo RANKL gene transfer exacerbated bone loss in WDFY3 conditional knockout mice, as evidenced by elevated serum TRAP, CTX-I and micro-CT analysis of distal femurs compared to wild type littermates. Taken together, our data highlight a novel role for WDFY3 in osteoclast development and function, which can be exploited for the treatment of musculoskeletal diseases.
KW - Autophagy
KW - Autophagy-linked FYVE containing protein
KW - Musculoskeletal diseases
KW - Osteoclast
KW - TRAF6
KW - WDFY3
UR - http://www.scopus.com/inward/record.url?scp=84977490443&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84977490443&partnerID=8YFLogxK
U2 - 10.1016/j.jaut.2016.06.004
DO - 10.1016/j.jaut.2016.06.004
M3 - Article
C2 - 27330028
AN - SCOPUS:84977490443
JO - Journal of Autoimmunity
JF - Journal of Autoimmunity
SN - 0896-8411
ER -