Dataset Information

Dissecting inflammatory response in intervertebral disc degeneration: triggering of CD44 by IL-1beta

ABSTRACT: Objectives/Design: Intervertebral Disc (IVD) degeneration has been associated with a chronic inflammatory response, but knowledge on the contribution of distinct IVD cells, namely CD44, to the progression of IVD degeneration remains elusive. Material: Bovine nucleus pulposus (NP) cells. Treatment: Stimulation with IL-1beta (10ng/ml). Methods: CD44 NP cells were sorted and compared by gene expression and proteomics with the negative counterpart. Dynamics of CD44 gene and protein expression was analyzed upon pro-inflammatory treatment. Results: CD44 has a multidimensional functional role in IVD metabolism, ECM synthesis and production of neuropermissive factors. CD44 widespread expression in NP has been partially associated with CD14 and CD45, resulting in the identification of distinct cell subsets. Conclusions: This study points out CD44 and CD44-based cell subsets as relevant targets in the modulation of the IVD pro-inflammatory/degenerative cascade.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Nucleus Pulposus

SUBMITTER: Hugo Osorio

LAB HEAD: Raquel M. Gonçalves

PROVIDER: PXD048373 | Pride | 2024-05-22

REPOSITORIES: Pride

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Dynamics of CD44<sup>+</sup> bovine nucleus pulposus cells with inflammation.

Ferreira J R JR Caldeira J J Sousa M M Barbosa M A MA Lamghari M M Almeida-Porada G G Gonçalves R M RM

Scientific reports 20240421 1

Intervertebral Disc (IVD) degeneration has been associated with a chronic inflammatory response, but knowledge on the contribution of distinct IVD cells, namely CD44, to the progression of IVD degeneration remains elusive. Here, bovine nucleus pulposus (NP) CD44 cells were sorted and compared by gene expression and proteomics with the negative counterpart. NP cells were then stimulated with IL-1b (10 ng/ml) and dynamics of CD44 gene and protein expression was analyzed upon pro-inflammatory treat ...[more]

PMID: 38644369

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Project description:Objective: Aging and early degeneration of the intervertebral disc (IVD) involves the substition of notochordal cells (NCs) in the nucleus pulposus (NP) by chondrocyte-like cells (CLCs). This study investigated the gene expression profiles involved in this process using NP tissue from both non-chondrodystrophic and chondrodystrophic dogs, a species with naturally occuring IVD degeneration. Methods: Dual channel DNA microarrays were used to compare 1) healthy NP tissue, 2) NP tissue with a mixed population of NCs and CLCs, and 3) NP tissue containing solely CLCs. Canonical Wnt-signaling was validated using qPCR of relevant Wnt target genes. Caveolin-1, a known regulator of canonical Wnt signaling, was investigated further in tissue sections using qPCR and immunohistochemistry, and in cultured NCs by qPCR and immunofluorescence. Also, the NP of 3-month-old caveolin-1 knock-out mice was histopathologically evaluated and compared with wild type mice of the same age. Results: Early IVD degeneration involved significant regulations in numerous pathways, such as extracellular matrix remodeling, Bone Morphogenetic Protein- , and Wnt/beta-catenin-signaling. With regard to Wnt/beta-catenin signaling, axin2 gene expression was significantly higher in chondrodystrophic dogs compared with non-chondrodystrophic dogs. IVD degeneration involved significant downregulation of axin2 gene expression and caveolin-1 gene and protein expression. NCs showed abundant caveolin-1 expression in vivo and in vitro, whereas CLCs did not. The NP of 3-month-old WT mice were rich in viable NCs, whereas the NPs of 3-month-old caveolin-1 knock-out mice contained chondroid-like matrix with small, rounded cells, the majority of which showed morphological signs of apoptosis. Conclusions: Aging and the onset of degeneration of the IVD involve significant downregulation of canonical Wnt signaling and caveolin-1, which appears to be essential in the physiology and preservation of NCs. DNA microarrays were used to compare nucleus palposus (NP) tissue of healthy and chondrodystrophic individuals. Furthermore, the situation of the tissue was divided into three stages: NCR: notochordal cell(NC) rich; CR: tissue containing solely chondrocyte-like-cells (CLC) and T: tissue with a mixed population of NCs and CLCs. Comparisons were analysed on a 2-color platform against a common reference sample, consisting of a multitude of canine organs, including liver, spleen, kidney, lung, hart, intestine and bone.

			Action	DRS
	CD44_minus_DonorA.raw	Raw
	CD44_minus_DonorB.raw	Raw
	CD44_minus_DonorC.raw	Raw
	CD44_minus_DonorD.raw	Raw
	CD44_plus_DonorA.raw	Raw