Overexpression of TIMP-3 in Chondrocytes Produces Transient Reduction in Growth Plate Length but Permanently Reduces Adult Bone Quality and Quantity

Poulet, Blandine; Liu, Ke; Plumb, Darren A.; Vo, P; Shah, Mittal; Staines, Katherine; Sampson, Alexandra; Nakamura, Hiroyuki; Nagase, Hideaki; Carriero, Alessandra; Shefelbine, Sandra J.; Pitsillides, Andrew A.; Bou‐Gharios, George

doi:10.1371/journal.pone.0167971

Cited by 21 publications

(24 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In order to verify that [‐1A]TIMP‐3 exerts different effects compared with native, endogenous TIMP‐3, we extended the experiments to include native TIMP‐3. In contrast to transduction with [‐1A]TIMP‐3, native TIMP‐3 completely abrogated bone formation, consistent with our previous in vivo findings, in which overexpression of Col2a1‐driven TIMP‐3 resulted in significant reduction in bone mass . Similarly, expression of mRNA for alkaline phosphatase, osteocalcin, type I collagen, and RUNX‐2, reflecting osteogenic stimulation, was found to be elevated in [‐1A]TIMP‐3–transduced osteoblasts and suppressed by TIMP‐3 (Figure E).…”

Section: Resultssupporting

confidence: 90%

“…Evidence presented herein shows that STR/Ort mice expressing high levels of [‐1A]TIMP‐3, especially females, had significantly increased trabecular parameters and cortical bone thickness, compared with control mice and mice expressing low levels of [‐1A]TIMP‐3. We expected the opposite effect, since our previous studies demonstrated that high levels of WT TIMP‐3 overexpression in mouse cartilage led to lack of secondary ossification, and mice with intermediate levels of TIMP‐3 had reduced bone mass . It has already been shown that MMP deficiency (e.g., deficiencies of MMP‐9 and membrane type 1 MMP) can lead to developmental skeletal abnormalities .…”

Section: Discussionmentioning

confidence: 93%

“…Extracellular trafficking of TIMP‐3 is regulated by the competition between ECM binding and endocytosis via low‐density lipoprotein receptor–related protein 1 (LRP‐1) . It has previously been shown that modifications in TIMP‐3 expression could modify joint degeneration and that loss/gain of TIMP‐3 function leads to significant alteration in bone structure . In the present study, we have used a variant of TIMP‐3, named [‐1A]TIMP‐3, which harbors a single alanine addition to the amino‐terminal end, which was shown to inhibit aggrecanases, namely ADAMTS‐4/5, with much greater selectivity than other MMPs .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Targeted Inhibition of Aggrecanases Prevents Articular Cartilage Degradation and Augments Bone Mass in the STR/Ort Mouse Model of Spontaneous Osteoarthritis

Kanakis

Liu

Poulet

et al. 2019

Arthritis & Rheumatology

Self Cite

View full text Add to dashboard Cite

Objective. Cartilage destruction in osteoarthritis (OA) is mediated mainly by matrix metalloproteinases (MMPs) and ADAMTS. The therapeutic candidature of targeting aggrecanases has not yet been defined in joints in which spontaneous OA arises from genetic susceptibility, as in the case of the STR/Ort mouse, without a traumatic or loadinduced etiology. In addition, we do not know the long-term effect of aggrecanase inhibition on bone. We undertook this study to assess the potential aggrecanase selectivity of a variant of tissue inhibitor of metalloproteinases 3 (TIMP-3), called [-1A]TIMP-3, on spontaneous OA development and bone formation in STR/Ort mice.Methods. Using the background of STR/Ort mice, which develop spontaneous OA, we generated transgenic mice that overexpress [-1A]TIMP-3, either ubiquitously or conditionally in chondrocytes. [-1A]TIMP-3 has an extra alanine at the N-terminus that selectively inhibits ADAMTS but not MMPs. We analyzed a range of OA-related measures in all mice at age 40 weeks.Results. Mice expressing high levels of [-1A]TIMP-3 were protected against development of OA, while those expressing low levels were not. Interestingly, we also found that high levels of [-1A]TIMP-3 transgene overexpression resulted in increased bone mass, particularly in females. This regulation of bone mass was at least partly direct, as adult mouse primary osteoblasts infected with [-1A]TIMP-3 in vitro showed elevated rates of mineralization.Conclusion. The results provide evidence that [-1A]TIMP-3-mediated inhibition of aggrecanases can protect against cartilage degradation in a naturally occurring mouse model of OA, and they highlight a novel role that aggrecanase inhibition may play in increased bone mass.

show abstract

Section: Resultssupporting

confidence: 90%

Section: Discussionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Targeted Inhibition of Aggrecanases Prevents Articular Cartilage Degradation and Augments Bone Mass in the STR/Ort Mouse Model of Spontaneous Osteoarthritis

Kanakis

Liu

Poulet

et al. 2019

Arthritis & Rheumatology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Additionally, the MMP13 and ADAMTS5 are able to degrade COL II, lead to articular cartilage destruction and initiate osteoarthritis 53 - 56 . Meanwhile, TIMP3, a key anticatabolic factor and one of the endogenous inhibitors of MMP13 and ADAMTS5 57 - 59 , was also studied. The results showed that MoO 4 2- ions inhibited catabolic responses of co-cultured RCs indicative of downregulation of HIF-2α, MMP13, ADAMTS5 as well as upregulation of TIMP3.…”

Section: Discussionmentioning

confidence: 99%

3D printing of Mo-containing scaffolds with activated anabolic responses and bi-lineage bioactivities

Dang¹,

Wang²,

Li³

et al. 2018

Theranostics

View full text Add to dashboard Cite

When osteochondral tissues suffer from focal or degenerative lesions caused by trauma or disorders, it is a tough challenge to regenerate them because of the limited self-healing capacity of articular cartilage. In this study, a series of Mo-doped bioactive glass ceramic (Mo-BGC) scaffolds were prepared and then systematically characterized. The released MoO42- ions from 7.5Mo-BGC scaffolds played a vital role in regenerating articular cartilage and subchondral bone synchronously.Methods: The Mo-BGC scaffolds were fabricated through employing both a sol-gel method and 3D printing technology. SEM, EDS, HRTEM, XRD, ICPAES and mechanical strength tests were respectively applied to analyze the physicochemical properties of Mo-BGC scaffolds. The proliferation and differentiation of rabbit chondrocytes (RCs) and human bone mesenchymal stem cells (HBMSCs) cultured with dilute solutions of 7.5Mo-BGC powder extract were investigated in vitro. The co-culture model was established to explore the possible mechanism of stimulatory effects of MoO42- ions on the RCs and HBMSCs. The efficacy of regenerating articular cartilage and subchondral bone using 7.5Mo-BGC scaffolds was evaluated in vivo.Results: The incorporation of Mo into BGC scaffolds effectively enhanced the compressive strength of scaffolds owing to the improved surface densification. The MoO42- ions released from the 7.5Mo-BGC powders remarkably promoted the proliferation and differentiation of both RCs and HBMSCs. The MoO42- ions in the co-culture system significantly stimulated the chondrogenic differentiation of RCs and meanwhile induced the chondrogenesis of HBMSCs. The chondrogenesis stimulated by MoO42- ions happened through two pathways: 1) MoO42- ions elicited anabolic responses through activating the HIF-1α signaling pathway; 2) MoO42- ions inhibited catabolic responses and protected cartilage matrix from degradation. The in vivo study showed that 7.5Mo-BGC scaffolds were able to significantly promote cartilage/bone regeneration when implanted into rabbit osteochondral defects for 8 and 12 weeks, displaying bi-lineage bioactivities.Conclusion: The 3D-printed Mo-BGC scaffolds with bi-lineage bioactivities and activated anabolic responses could offer an effective strategy for cartilage/bone interface regeneration.

show abstract

“…Thus, we speculate that sustained TIMP-3 expression, and the consequent persistent inhibition of ADAM10, as well as the inhibition of other metalloproteinases, can have detrimental effects in vivo . In support of this hypothesis, transgenic mice overexpressing TIMP-3 in chondrocytes display abnormalities in bone development, including decreased osteoblast differentiation and reduction in growth plate length and bone mass 52 , 53 . Such a phenotype may be due to dysregulation of Notch signaling, which plays a crucial role in maintaining bone homeostasis by regulating proliferation and differentiation of both osteoblasts and osteoclasts 54 .…”

Section: Discussionmentioning

confidence: 86%

Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor

Scilabra

Pigoni

Pravatà

et al. 2018

Sci Rep

View full text Add to dashboard Cite

The tissue inhibitor of metalloproteinases-3 (TIMP-3) is a major regulator of extracellular matrix turnover and protein shedding by inhibiting different classes of metalloproteinases, including disintegrin metalloproteinases (ADAMs). Tissue bioavailability of TIMP-3 is regulated by the endocytic receptor low-density-lipoprotein receptor-related protein-1 (LRP-1). TIMP-3 plays protective roles in disease. Thus, different approaches have been developed aiming to increase TIMP-3 bioavailability, yet overall effects of increased TIMP-3 in vivo have not been investigated. Herein, by using unbiased mass-spectrometry we demonstrate that TIMP-3-overexpression in HEK293 cells has a dual effect on shedding of transmembrane proteins and turnover of soluble proteins. Several membrane proteins showing reduced shedding are known as ADAM10 substrates, suggesting that exogenous TIMP-3 preferentially inhibits ADAM10 in HEK293 cells. Additionally identified shed membrane proteins may be novel ADAM10 substrate candidates. TIMP-3-overexpression also increased extracellular levels of several soluble proteins, including TIMP-1, MIF and SPARC. Levels of these proteins similarly increased upon LRP-1 inactivation, suggesting that TIMP-3 increases soluble protein levels by competing for their binding to LRP-1 and their subsequent internalization. In conclusion, our study reveals that increased levels of TIMP-3 induce substantial modifications in the cellular secretome and that TIMP-3-based therapies may potentially provoke undesired, dysregulated functions of ADAM10 and LRP-1.

show abstract

Overexpression of TIMP-3 in Chondrocytes Produces Transient Reduction in Growth Plate Length but Permanently Reduces Adult Bone Quality and Quantity

Cited by 21 publications

References 41 publications

Targeted Inhibition of Aggrecanases Prevents Articular Cartilage Degradation and Augments Bone Mass in the STR/Ort Mouse Model of Spontaneous Osteoarthritis

Targeted Inhibition of Aggrecanases Prevents Articular Cartilage Degradation and Augments Bone Mass in the STR/Ort Mouse Model of Spontaneous Osteoarthritis

3D printing of Mo-containing scaffolds with activated anabolic responses and bi-lineage bioactivities

Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor

Contact Info

Product

Resources

About