Endogenous CCN family member WISP1 inhibits trauma-induced heterotopic ossification
Ginny Ching-Yun Hsu, Simone Marini, Stefano Negri, Yiyun Wang, Jiajia Xu, Chase Pagani, Charles Hwang, David Stepien, Carolyn A. Meyers, Sarah Miller, Edward McCarthy, Karen M. Lyons, Benjamin Levi, Aaron W. James
Ginny Ching-Yun Hsu, Simone Marini, Stefano Negri, Yiyun Wang, Jiajia Xu, Chase Pagani, Charles Hwang, David Stepien, Carolyn A. Meyers, Sarah Miller, Edward McCarthy, Karen M. Lyons, Benjamin Levi, Aaron W. James
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Research Article Bone biology Stem cells

Endogenous CCN family member WISP1 inhibits trauma-induced heterotopic ossification

Abstract

Heterotopic ossification (HO) is defined as abnormal differentiation of local stromal cells of mesenchymal origin, resulting in pathologic cartilage and bone matrix deposition. Cyr61, CTGF, Nov (CCN) family members are matricellular proteins that have diverse regulatory functions on cell proliferation and differentiation, including the regulation of chondrogenesis. However, little is known regarding CCN family member expression or function in HO. Here, a combination of bulk and single-cell RNA sequencing defined the dynamic temporospatial pattern of CCN family member induction within a mouse model of trauma-induced HO. Among CCN family proteins, Wisp1 (also known as Ccn4) was most upregulated during the evolution of HO, and Wisp1 expression corresponded with chondrogenic gene profile. Immunohistochemistry confirmed WISP1 expression across traumatic and genetic HO mouse models as well as in human HO samples. Transgenic Wisp1LacZ/LacZ knockin animals showed an increase in endochondral ossification in HO after trauma. Finally, the transcriptome of Wisp1-null tenocytes revealed enrichment in signaling pathways, such as the STAT3 and PCP signaling pathways, that may explain increased HO in the context of Wisp1 deficiency. In sum, CCN family members, and in particular Wisp1, are spatiotemporally associated with and negatively regulate trauma-induced HO formation.

Authors

Ginny Ching-Yun Hsu, Simone Marini, Stefano Negri, Yiyun Wang, Jiajia Xu, Chase Pagani, Charles Hwang, David Stepien, Carolyn A. Meyers, Sarah Miller, Edward McCarthy, Karen M. Lyons, Benjamin Levi, Aaron W. James

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Figure 1

Bulk total RNA sequencing identifies Wisp1 enrichment in posttraumatic heterotopic ossification.

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Bulk total RNA sequencing identifies Wisp1 enrichment in posttraumatic h...
Bulk RNA sequencing of Achilles tendon tenotomy site. Tissue was collected from the soft tissue around the injury site from the posterior compartment between the muscular origin and calcaneal insertion of Achilles tendon. Corresponding soft tissue, including intact Achilles tendon, was also harvested from the uninjured contralateral hind limb. In each image, data are presented in volcano plots and logarithmic fold change is shown on the x axis (log2). The y axis depicts P value (log10). Data were derived from n = 3 pooled samples per group, with expression examined at 3 weeks after injury. (A) CCN family members, including Ctgf, Cyr61, Nov, Wisp1, Wisp2, and Wisp3. (B) Selective cartilage-associated genes. (C) Selective bone-associated genes. All analyses were performed on the same data set. Light red and dark red dots indicate genes with a >2 SD increase and a >3 SD increase in comparison with uninjured control, respectively. Light blue and dark blue dots indicate genes with a >2 SD decrease and a >3 SD decrease in comparison with uninjured control, respectively.