Articular cartilage matrix vesicles (ACVs) are small membrane-bound extracellular organelles found in normal cartilage and chondrocyte cultures. To date, they have almost exclusively been described in reference to their role in pathologic matrix mineralization in articular cartilage. Yet, their presence in normal cartilage and roles for ACVs other than mineral formation remain unexplored. We recently demonstrated that ACVs contain functional RNA and are able to transfer labeled RNA and protein to naove chondrocytes. The addition of 1-10 5g/ml normal ACVs to normal chondrocyte monolayers increased markers characteristic of the hypertrophic chondrocytes seen in osteoarthritic (OA) cartilage. These exciting findings suggest that ACVs directly interact with chondrocytes and may participate in the cellular changes seen in OA. Preliminary data suggest an important role for proteoglycans in binding ACVs to pericellular matrix. We hypothesize that increased activity of proteoglycan-degrading enzymes in early OA promotes ACV "mobility" and fosters ACV-chondrocyte interactions in cartilage. However, the mechanisms through which ACVs bind to matrix components and the identity of the factors modulating matrix binding are unstudied. Similarly, little is known about the factors that mediate the cellular effects of ACVs. Exposure of normal articular chondrocytes to exogenous transglutaminase (Tgase) enzymes promotes chondrocyte hypertrophy in an integrin-dependent manner. As Tgases are present in high levels in ACVs, we hypothesize that Tgases in or on ACVs are responsible for inducing the hypertrophic phenotype in chondrocytes exposed to ACVs and that this effect is integrin-dependent. Using porcine ACV and chondrocytes, and replicating key experiments with ACVs from purchased human chondrocytes, we will investigate the following hypotheses:
Specific aim 1 : To investigate the hypothesis that ACVs bind to proteoglycans in matrix and ADAMTS4 and 5 release ACVs from cartilage matrix.
Specific aim 2 : To investigate the hypothesis that Tgases in ACVs are responsible for induction of the hypertrophic phenotype seen in chondrocytes after ACV exposure and that this is integrin-dependent. The ultimate goal of this work is to understand the role of ACVs in cartilage in the hope that manipulating the contents or availability of these organelles might contribute to the treatment of OA.

Public Health Relevance

Narrative: Osteoarthritis is the most common form of arthritis in adults and affects 40 million Americans. It is currently untreatable, and often results in significant loss of independence and increased medical costs for affected people. Risk factors for OA, including advanced age, prior trauma, and obesity are particularly prevalent in the US veteran population and OA is a leading source of disability in veterans. This proposal deals with the role of small extracellular vesicles, called articular cartilage vesicles (ACVs) in OA. ACVs carry RNA and protein that can be transferred to chondrocytes. Transfer of ACV contents to chondrocytes mimics changes seen in OA. Understanding the role of ACVs in cartilage and ultimately manipulating the contents of ACVs could lead to novel therapies for OA.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX000812-02
Application #
8198369
Study Section
Immunology A (IMMA)
Project Start
2010-10-01
Project End
2014-09-30
Budget Start
2011-10-01
Budget End
2012-09-30
Support Year
2
Fiscal Year
2012
Total Cost
Indirect Cost
Name
Clement J. Zablocki VA Medical Center
Department
Type
DUNS #
078952454
City
Milwaukee
State
WI
Country
United States
Zip Code
53295