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A functional SNP in CILP, encoding cartilage intermediate layer protein, is associated with susceptibility to lumbar disc disease


Lumbar disc disease (LDD) is caused by degeneration of intervertebral discs of the lumbar spine. One of the most common musculoskeletal disorders1, LDD has strong genetic determinants2,3,4. Using a case-control association study, we identified a functional SNP (1184T → C, resulting in the amino acid substitution I395T) in CILP, which encodes the cartilage intermediate layer protein, that acts as a modulator of LDD susceptibility. CILP was expressed abundantly in intervertebral discs, and its expression increased as disc degeneration progressed. CILP colocalized with TGF-β1 in clustering chondrocytes and their territorial matrices in intervertebral discs. CILP inhibited TGF-β1–mediated induction of cartilage matrix genes through direct interaction with TGF-β1 and inhibition of TGF-β1 signaling. The susceptibility-associated 1184C allele showed increased binding and inhibition of TGF-β1. Therefore, we conclude that the extracellular matrix protein CILP regulates TGF-β signaling and that this regulation has a crucial role in the etiology and pathogenesis of LDD. Our study also adds to the list of connective tissue diseases that are associated with TGF-β.

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Figure 1: LD mapping of the susceptibility gene for LDD.
Figure 2: Effects of the LDD-associated SNP (1184T → C) in CILP on TGF-β1–induced expression of aggrecan (a) and type II collagen (b) in rabbit nucleus pulposus cells.
Figure 3: Suppression of the TGF-β signal by CILP in nucleus pulposus cells.
Figure 4: Binding of CILP to TGF-β1 in vitro.
Figure 5: Localization of CILP and TGF-β1 in human lumbar disc tissue.

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We thank the affected individuals for participating in this study; H. Hirabayashi, T. Kono, N. Hosogane, H. Hase, T. Ogura, T. Tanaka, M. Sasahara, A. Sano, H. Ishihara, M. Kanamori and K. Toyoshima for assistance; and Y. Takanashi and T. Matsushima for technical assistance.

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Correspondence to Shiro Ikegawa.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Real-time RT-PCR analysis of CILP mRNA in various human tissues and cell lines. (PDF 86 kb)

Supplementary Fig. 2

CILP expression in intervertebral disc tissue from LDD patients. (PDF 62 kb)

Supplementary Fig. 3

CILP expression in intervertebral disc tissues from a normal subject and an LDD patient (Schneiderman's grade 3). (PDF 61 kb)

Supplementary Fig. 4

Lack of NTPPHase activity in COS-7 cells expressing F-CILP. (PDF 75 kb)

Supplementary Fig. 5

Purified F-CILP containing p.I395 or p.T395, visualized by silver staining. (PDF 75 kb)

Supplementary Fig. 6

Effects of the LDD-associated SNP (p.I395T) in N-CILP on TGF-β1-induced expression of the aggrecan and type II collagen genes in rabbit nucleus pulposus cells. (PDF 94 kb)

Supplementary Table 1

Assessment of population stratification. (PDF 61 kb)

Supplementary Table 2

Haplotype analysis of CILP with lumbar disc herniation. (PDF 96 kb)

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Seki, S., Kawaguchi, Y., Chiba, K. et al. A functional SNP in CILP, encoding cartilage intermediate layer protein, is associated with susceptibility to lumbar disc disease. Nat Genet 37, 607–612 (2005).

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