Role of HTRA1 in bone formation and regeneration: In vitro and in vivo evaluation

The role of mammalian high temperature requirement protease A1 (HTRA1) in somatic stem cell differentiation and mineralized matrix formation remains controversial, having been demonstrated to impart either anti- or pro-osteogenic effects, depending on the in vitro cell model used. The aim of this study was therefore to further evaluate the role of HTRA1 in regulating the differentiation potential and lineage commitment of murine mesenchymal stem cells in vitro, and to assess its influence on bone structure and regeneration in vivo. Our results demonstrated that short hairpin RNA-mediated ablation of Htra1 in the murine mesenchymal cell line C3H10T1/2 increased the expression of several osteogenic gene markers, and significantly enhanced matrix mineralization in response to BMP-2 stimulation. These effects were concomitant with decreases in the expression of chondrogenic gene markers, and increases in adipogenic gene expression and lipid accrual. Despite the profound effects of loss-of-function of HTRA1 on this in vitro osteochondral model, these were not reproduced in vivo, where bone microarchitecture and regeneration in 16-week-old Htra1-knockout mice remained unaltered as compared to wild-type controls. By comparison, analysis of femurs from 52-week-old mice revealed that bone structure was better preserved in Htra1-knockout mice than age-matched wild-type controls. These findings therefore provide additional insights into the role played by HTRA1 in regulating mesenchymal stem cell differentiation, and offer opportunities for improving our understanding of how this multifunctional protease may act to influence bone quality.

[1]  Jack C. Yu,et al.  Effects of In Utero Thyroxine Exposure on Murine Cranial Suture Growth , 2016, PloS one.

[2]  G. Onder,et al.  Association of frailty with the serine protease HtrA1 in older adults , 2016, Experimental Gerontology.

[3]  M. Kawaichi,et al.  Epigenetic silencing of serine protease HTRA1 drives polyploidy , 2016, BMC Cancer.

[4]  M. Blüher,et al.  Novel Function of Serine Protease HTRA1 in Inhibiting Adipogenic Differentiation of Human Mesenchymal Stem Cells via MAP Kinase‐Mediated MMP Upregulation , 2016, Stem cells.

[5]  P. Richards,et al.  Loss-of-Function of HtrA1 Abrogates All-Trans Retinoic Acid-Induced Osteogenic Differentiation of Mouse Adipose-Derived Stromal Cells Through Deficiencies in p70S6K Activation. , 2016, Stem cells and development.

[6]  Mingzhi Zhao,et al.  Serum HtrA1 is differentially regulated between early-onset and late-onset preeclampsia. , 2015, Placenta.

[7]  A. Joachimiak,et al.  Structural and Functional Analysis of Human HtrA3 Protease and Its Subdomains , 2015, PloS one.

[8]  D. Marzioni,et al.  HtrA1: Its future potential as a novel biomarker for cancer , 2015, Oncology reports.

[9]  Ningli Wang,et al.  Inhibition of cell proliferation and migration after HTRA1 knockdown in retinal pigment epithelial cells , 2015, Graefe's Archive for Clinical and Experimental Ophthalmology.

[10]  Ran Li,et al.  HtrA1 may regulate the osteogenic differentiation of human periodontal ligament cells by TGF-β1 , 2015, Journal of Molecular Histology.

[11]  D. Werring,et al.  Cerebral small vessel disease-related protease HtrA1 processes latent TGF-β binding protein 1 and facilitates TGF-β signaling , 2014, Proceedings of the National Academy of Sciences.

[12]  R. O’Keefe,et al.  The Convergence of Fracture Repair and Stem Cells: Interplay of Genes, Aging, Environmental Factors and Disease , 2014, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[13]  V. Rosen,et al.  HtrA1 is upregulated during RANKL‐induced osteoclastogenesis, and negatively regulates osteoblast differentiation and BMP2‐induced Smad1/5/8, ERK and p38 phosphorylation , 2014, FEBS letters.

[14]  J. Graham,et al.  Serine Protease HTRA1 Antagonizes Transforming Growth Factor-β Signaling by Cleaving Its Receptors and Loss of HTRA1 In Vivo Enhances Bone Formation , 2013, PloS one.

[15]  Sung-Cheng Huang,et al.  PPARγ silencing enhances osteogenic differentiation of human adipose-derived mesenchymal stem cells , 2013, Journal of cellular and molecular medicine.

[16]  T. He,et al.  BMP signaling in mesenchymal stem cell differentiation and bone formation. , 2013, Journal of biomedical science and engineering.

[17]  M. Kawaichi,et al.  HtrA1 is induced by oxidative stress and enhances cell senescence through p38 MAPK pathway. , 2013, Experimental eye research.

[18]  P. Richards,et al.  The emerging roles of HTRA1 in musculoskeletal disease. , 2013, The American journal of pathology.

[19]  M. Ehrmann,et al.  Human Serine Protease HTRA1 Positively Regulates Osteogenesis of Human Bone Marrow‐derived Mesenchymal Stem Cells and Mineralization of Differentiating Bone‐forming Cells Through the Modulation of Extracellular Matrix Protein , 2012, Stem cells.

[20]  Cheol‐Hee Kim,et al.  HtrA1 Is a Novel Antagonist Controlling Fibroblast Growth Factor (FGF) Signaling via Cleavage of FGF8 , 2012, Molecular and Cellular Biology.

[21]  M. van Lookeren Campagne,et al.  Structural and functional analysis of HtrA1 and its subdomains. , 2012, Structure.

[22]  T. Liebscher,et al.  Detrimental Role for Human High Temperature Requirement Serine Protease A1 (HTRA1) in the Pathogenesis of Intervertebral Disc (IVD) Degeneration* , 2012, The Journal of Biological Chemistry.

[23]  R. Huber,et al.  Human High Temperature Requirement Serine Protease A1 (HTRA1) Degrades Tau Protein Aggregates* , 2012, The Journal of Biological Chemistry.

[24]  Amrita,et al.  Increased expression of multifunctional serine protease, HTRA1, in retinal pigment epithelium induces polypoidal choroidal vasculopathy in mice , 2011, Proceedings of the National Academy of Sciences.

[25]  R. Huber,et al.  HTRA proteases: regulated proteolysis in protein quality control , 2011, Nature Reviews Molecular Cell Biology.

[26]  K. Arima,et al.  Cerebral small-vessel disease protein HTRA1 controls the amount of TGF-β1 via cleavage of proTGF-β1. , 2011, Human molecular genetics.

[27]  À. Rovira,et al.  A missense HTRA1 mutation expands CARASIL syndrome to the Caucasian population , 2010, Neurology.

[28]  M. Caraglia,et al.  The Serine Protease HtrA1 Specifically Interacts and Degrades the Tuberous Sclerosis Complex 2 Protein , 2010, Molecular Cancer Research.

[29]  Ralph Müller,et al.  Guidelines for assessment of bone microstructure in rodents using micro–computed tomography , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[30]  Keita Ito,et al.  Influence of defective bone marrow osteogenesis on fracture repair in an experimental model of senile osteoporosis , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[31]  Keita Ito,et al.  Fixation compliance in a mouse osteotomy model induces two different processes of bone healing but does not lead to delayed union. , 2009, Journal of biomechanics.

[32]  J. Chien,et al.  Serine Protease HtrA1 Associates with Microtubules and Inhibits Cell Migration , 2009, Molecular and Cellular Biology.

[33]  T. He,et al.  A comprehensive analysis of the dual roles of BMPs in regulating adipogenic and osteogenic differentiation of mesenchymal progenitor cells. , 2009, Stem cells and development.

[34]  L. Dauphinot,et al.  HtrA1-dependent proteolysis of TGF-β controls both neuronal maturation and developmental survival , 2008, Cell Death and Differentiation.

[35]  R. Boot-Handford,et al.  HtrA1 Inhibits Mineral Deposition by Osteoblasts , 2008, Journal of Biological Chemistry.

[36]  Ralph Müller,et al.  Automated compartmental analysis for high-throughput skeletal phenotyping in femora of genetic mouse models. , 2007, Bone.

[37]  N. Camp,et al.  A Variant of the HTRA1 Gene Increases Susceptibility to Age-Related Macular Degeneration , 2006, Science.

[38]  C. Barnstable,et al.  HTRA1 Promoter Polymorphism in Wet Age-Related Macular Degeneration , 2006, Science.

[39]  L. Salamonsen,et al.  Serine proteases HTRA1 and HTRA3 are down-regulated with increasing grades of human endometrial cancer. , 2006, Gynecologic oncology.

[40]  Tim Clausen,et al.  The Role of Human HtrA1 in Arthritic Disease* , 2006, Journal of Biological Chemistry.

[41]  M. Kawaichi,et al.  Expression of mouse HtrA1 serine protease in normal bone and cartilage and its upregulation in joint cartilage damaged by experimental arthritis. , 2005, Bone.

[42]  Alfonso Baldi,et al.  Implications of the serine protease HtrA1 in amyloid precursor protein processing , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[43]  R. Tuan,et al.  Transdifferentiation potential of human mesenchymal stem cells derived from bone marrow , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[44]  M. Kawaichi,et al.  Developmentally regulated expression of mouse HtrA3 and its role as an inhibitor of TGF‐β signaling , 2004, Development, growth & differentiation.

[45]  Kozo Nakamura,et al.  PPARgamma insufficiency enhances osteogenesis through osteoblast formation from bone marrow progenitors. , 2004, The Journal of clinical investigation.

[46]  M. Kawaichi,et al.  HtrA1 serine protease inhibits signaling mediated by Tgfβ family proteins , 2004, Development.

[47]  David I. Smith,et al.  A candidate tumor suppressor HtrA1 is downregulated in ovarian cancer , 2004, Oncogene.

[48]  Xian-Yang Zhang,et al.  Transduction of Bone-Marrow-Derived Mesenchymal Stem Cells by Using Lentivirus Vectors Pseudotyped with Modified RD114 Envelope Glycoproteins , 2004, Journal of Virology.

[49]  T. Einhorn,et al.  BMP treatment of C3H10T1/2 mesenchymal stem cells induces both chondrogenesis and osteogenesis , 2003, Journal of cellular biochemistry.

[50]  B. Scammell,et al.  The fate of soft callus chondrocytes during long bone fracture repair , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[51]  C. Southan,et al.  The HtrA family of proteases: implications for protein composition and cell fate. , 2002, Molecular cell.

[52]  H F Sailer,et al.  Slow and continuous application of human recombinant bone morphogenetic protein via biodegradable poly(lactide-co-glycolide) foamspheres. , 2002, International journal of oral and maxillofacial surgery.

[53]  Theodore Miclau,et al.  Does adult fracture repair recapitulate embryonic skeletal formation? , 1999, Mechanisms of Development.

[54]  S. Karlsson,et al.  Transduction of nondividing cells using pseudotyped defective high-titer HIV type 1 particles. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[55]  C. Heidelberger,et al.  Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. , 1973, Cancer research.

[56]  M. Kawaichi,et al.  Abnormal development of placenta in HtrA1-deficient mice. , 2015, Developmental biology.