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Investigating Global Changes in Gene Expression in a Murine Model of Cherubism

ABSTRACT: Investigating Global Changes in Gene Expression in a Murine Model of Cherubism

PROVIDER: PRJNA579596 | ENA |

REPOSITORIES: ENA

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Investigating Global Changes in Gene Expression in a Murine Model of Cherubism

Project description:Purpose: To identify the mechanistic changes that lead to impaired osteogenesis and spontaneous osteoclast formation in both the hematopoietic and mesenchymal cell populations the makeup bone marrow cultures derived from cherubism mice (Sh3bp2KI/KI). Methods: Bone marrow cultures derived from wild type and cherubism mice were grown for seven days. On day seven, hematopoietic and mesenchymal cell types were sorted using CD45 and Sca1 cell surface markers. CD45+Sca1+ identified the hematopoietic population. CD45-Sca1+ identified the mesenchymal population. Total RNA was prepared from cell sorted populations. RNA quality was confirmed, ribosomal RNAs were depleted, libraries were prepared, and paired-end sequencing was performed on a illumina platform. We performed this study using six biological replicates. Results: Gene expression differences were identified Conclusions: Our study, for the first time, identifies the impact of a cherubism mutation on the global transcriptome of hematopoietic and mesenchymal cells extracted from murine bone marrow stromal cultures. With this study, we have found novel molecular signatures that are consistent with the cherubism phenotype (inflammation, bone loss, and fibrosis).

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Investigating global gene expression changes in a murine model of cherubism.

Project description:Cherubism is a rare genetic disorder caused primarily by mutations in SH3BP2 resulting in excessive bone resorption and fibrous tissue overgrowth in the lower portions of the face. Bone marrow derived cell cultures derived from a murine model of cherubism display poor osteogenesis and spontaneous osteoclast formation. To develop a deeper understanding for the potential underlying mechanisms contributing to these phenotypes in mice, we compared global gene expression changes in hematopoietic and mesenchymal cell populations between cherubism and wild type mice. In the hematopoietic population, not surprisingly, upregulated genes were significantly enriched for functions related to osteoclastogenesis. However, these upregulated genes were also significantly enriched for functions associated with inflammation including arachidonic acid/prostaglandin signaling, regulators of coagulation and autoinflammation, extracellular matrix remodeling, and chemokine expression. In the mesenchymal population, we observed down regulation of osteoblast and adventitial reticular cell marker genes. Regulators of BMP and Wnt pathway associated genes showed numerous changes in gene expression, likely implicating the down regulation of BMP signaling and possibly the activation of certain Wnt pathways. Analyses of the cherubism derived mesenchymal population also revealed interesting changes in gene expression related to inflammation including the expression of distinct granzymes, chemokines, and sulfotransferases. These studies reveal complex changes in gene expression elicited from a cherubic mutation in Sh3bp2 that are informative to the mechanisms responding to inflammatory stimuli and repressing osteogenesis. The outcomes of this work are likely to have relevance not only to cherubism, but other inflammatory conditions impacting the skeleton.

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