Lentiviral small hairpin RNA knockdown of macrophage inflammatory protein-1γ ameliorates experimentally induced osteoarthritis in mice

Po Chuan Shen, Chia Sing Lu, Ai-Li Shiau, Che Hsin Lee, I. Ming Jou, Jeng Long Hsieh

Research output: Contribution to journalArticle

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Abstract

Immune cells are involved in the pathogenesis of osteoarthritis (OA). CD4+ T cells were activated during the onset of OA and induced macrophage inflammatory protein (MIP)-1γ expression and subsequent osteoclast formation. We evaluated the effects of local knockdown of MIP-1γ in a mouse OA model induced by anterior cruciate ligament transection. The mouse macrophage cell lines and osteoclast-like cells generated from immature hematopoietic monocyte/macrophage progenitors of murine bone marrow were cocultured with either receptor activator of NFκB ligand (RANKL) or CD4+ T cells. The levels of MIP-1γ and RANKL in cells and mice were examined by enzyme-linked immunosorbent assay (ELISA). The osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase and cathepsin K staining. OA was induced in one hind-leg knee joint of B6 mice. Lentiviral vector encoding MIP-1γ small hairpin RNA (shRNA) and control vector were individually injected intra-articularly into the knee joints, which were histologically assessed for manifestations of OA. The expression of MIP-1γ and matrix metalloproteinase (MMP)-13 and the infiltration of CD4+ T cells, macrophages, and osteoclastogenesis in tissues were examined using immunohistochemistry. CD4+ T cells were involved in OA by inducing MIP-1γ expression in osteoclast progenitors and the subsequent osteoclast formation. Neutralizing MIP-1γ with a specific antibody abolishes RANKL-stimulated and CD4+ T-cell-stimulated osteoclast formation. MIP-1γ levels were significantly higher in synovium and the chondro-osseous junction of joints 90 days postsurgery. The number of infiltrated CD4+ T cells and macrophages and IL-1β expression were reduced in the synovial tissues of mice treated with MIP-1γ shRNA. Histopathological examinations revealed that mice treated with MIP-1γ shRNA had less severe OA than control mice had, as well as decreased osteoclast formation and MMP-13 expression. Locally inhibiting MIP-1γ expression may ameliorate disease progression and provide a new OA therapy.

Original languageEnglish
Pages (from-to)871-882
Number of pages12
JournalHuman Gene Therapy
Volume24
Issue number10
DOIs
Publication statusPublished - 2013 Oct 1

Fingerprint

Macrophage Inflammatory Proteins
Osteoarthritis
Small Interfering RNA
Osteoclasts
T-Lymphocytes
Macrophages
Matrix Metalloproteinase 13
Knee Joint
Osteogenesis
Cathepsin K
Synovial Membrane
Anterior Cruciate Ligament
Interleukin-1
Disease Progression
Monocytes
Leg
Joints
Bone Marrow
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Shen, Po Chuan ; Lu, Chia Sing ; Shiau, Ai-Li ; Lee, Che Hsin ; Jou, I. Ming ; Hsieh, Jeng Long. / Lentiviral small hairpin RNA knockdown of macrophage inflammatory protein-1γ ameliorates experimentally induced osteoarthritis in mice. In: Human Gene Therapy. 2013 ; Vol. 24, No. 10. pp. 871-882.
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Lentiviral small hairpin RNA knockdown of macrophage inflammatory protein-1γ ameliorates experimentally induced osteoarthritis in mice. / Shen, Po Chuan; Lu, Chia Sing; Shiau, Ai-Li; Lee, Che Hsin; Jou, I. Ming; Hsieh, Jeng Long.

In: Human Gene Therapy, Vol. 24, No. 10, 01.10.2013, p. 871-882.

Research output: Contribution to journalArticle

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T1 - Lentiviral small hairpin RNA knockdown of macrophage inflammatory protein-1γ ameliorates experimentally induced osteoarthritis in mice

AU - Shen, Po Chuan

AU - Lu, Chia Sing

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AU - Jou, I. Ming

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