Functional Insights in PLS3-Mediated Osteogenic Regulation
Wenchao Zhong,
Janine Neugebauer,
Janak L. Pathak,
Xingyang Li,
Gerard Pals,
M. Carola Zillikens,
Elisabeth M. W. Eekhoff,
Nathalie Bravenboer,
Qingbin Zhang,
Matthias Hammerschmidt,
Brunhilde Wirth,
Dimitra Micha
Affiliations
Wenchao Zhong
Department of Human Genetics, Amsterdam UMC Location Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
Janine Neugebauer
Institute of Human Genetics University Hospital of Cologne, University of Cologne, 50931 Cologne, Germany
Janak L. Pathak
Department of Temporomandibular Joint, School and Hospital of Stomatology, Guangdong Engineering Research Center of Oral Restoration and Reconstruction & Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou Medical University, Guangzhou 510060, China
Xingyang Li
Department of Temporomandibular Joint, School and Hospital of Stomatology, Guangdong Engineering Research Center of Oral Restoration and Reconstruction & Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou Medical University, Guangzhou 510060, China
Gerard Pals
Department of Human Genetics, Amsterdam UMC Location Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
M. Carola Zillikens
Department of Internal Medicine, Erasmus MC, University Medical Center Rotterdam, 3015 CE Rotterdam, The Netherlands
Elisabeth M. W. Eekhoff
Amsterdam Movement Sciences, Tissue Function And Regeneration, 1081 HV Amsterdam, The Netherlands
Nathalie Bravenboer
Department of Clinical Chemistry, Amsterdam UMC Location Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
Qingbin Zhang
Department of Temporomandibular Joint, School and Hospital of Stomatology, Guangdong Engineering Research Center of Oral Restoration and Reconstruction & Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou Medical University, Guangzhou 510060, China
Matthias Hammerschmidt
Developmental Biology Unit, Institute of Zoology, University of Cologne, 50931 Cologne, Germany
Brunhilde Wirth
Institute of Human Genetics University Hospital of Cologne, University of Cologne, 50931 Cologne, Germany
Dimitra Micha
Department of Human Genetics, Amsterdam UMC Location Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
Plastin-3 (PLS3) encodes T-plastin, an actin-bundling protein mediating the formation of actin filaments by which numerous cellular processes are regulated. Loss-of-function genetic defects in PLS3 are reported to cause X-linked osteoporosis and childhood-onset fractures. However, the molecular etiology of PLS3 remains elusive. Functional compensation by actin-bundling proteins ACTN1, ACTN4, and FSCN1 was investigated in zebrafish following morpholino-mediated pls3 knockdown. Primary dermal fibroblasts from six patients with a PLS3 variant were also used to examine expression of these proteins during osteogenic differentiation. In addition, Pls3 knockdown in the murine MLO-Y4 cell line was employed to provide insights in global gene expression. Our results showed that ACTN1 and ACTN4 can rescue the skeletal deformities in zebrafish after pls3 knockdown, but this was inadequate for FSCN1. Patients’ fibroblasts showed the same osteogenic transdifferentiation ability as healthy donors. RNA-seq results showed differential expression in Wnt1, Nos1ap, and Myh3 after Pls3 knockdown in MLO-Y4 cells, which were also associated with the Wnt and Th17 cell differentiation pathways. Moreover, WNT2 was significantly increased in patient osteoblast-like cells compared to healthy donors. Altogether, our findings in different bone cell types indicate that the mechanism of PLS3-related pathology extends beyond actin-bundling proteins, implicating broader pathways of bone metabolism.
