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Relationship: 2685
Title
Decrease, sox9 expression leads to Smaller and morphologically distorted facial cartilage structures
Upstream event
Downstream event
Key Event Relationship Overview
AOPs Referencing Relationship
AOP Name | Adjacency | Weight of Evidence | Quantitative Understanding | Point of Contact | Author Status | OECD Status |
---|---|---|---|---|---|---|
Aryl hydrocarbon receptor activation leading to early life stage mortality via sox9 repression induced impeded craniofacial development | adjacent | High | Low | Prarthana Shankar (send email) | Under development: Not open for comment. Do not cite | Under Review |
Taxonomic Applicability
Sex Applicability
Sex | Evidence |
---|---|
Unspecific | High |
Life Stage Applicability
Term | Evidence |
---|---|
Embryo | High |
Development | High |
Key Event Relationship Description
- Sox9 is an important transcriptional regulator that has been implicated in several functions including craniofacial development, specifically via chondrogenesis or the formation of cartilage structure (Lefebvre and Dvir-Ginzberg 2017).
- Additionally, exposure of different animals to relevant environmental pollutants leads to a significant decrease of sox9 expression (Garcia et al., 2017; Shi et al., 2017; Tussellino et al., 2016).
- This KER provides lines of evidence linking the sox9 repression to alterations in craniofacial development and function.
Evidence Collection Strategy
Evidence Supporting this KER
KER 2685 concordance table: https://aopwiki.org/system/dragonfly/production/2022/10/20/71wqxtrj9g_Concordance_Table_sox9_to_craniofacial_clean.pdf
Biological Plausibility
- Compelling biological plausibility evidence comes from studies in multiple species showing the spatiotemporal expression of sox9 in the developing cartilage structures of the jaw.
- In mice, sox9 mRNA is widely expressed in the condylar anlage and Meckel’s cartilage (Shibata et al. 2006), and the sox9 protein in the tissue layer of secondary cartilage (Hirouchi et al., 2018; Zhang et al., 2013) suggesting roles of sox9 in chondrogenesis. Additionally, sox9 is expressed widely during palatogenesis (Nie 2006; Watanabe et al., 2016), and is also found in the temporomandibular joint of developing mice (TMJ) (Wang et al., 2011). There is a some evidence for sox9 being expressed in the condyle cartilage, as well as the proliferative layer and in the chondrocytes of developing rats (Al-Dujaili et al., 2018; Rabie and Hägg 2002).
- Sox9 is expressed within the developing chondrocytes of rabbits (Huang et al., 2015).
- Sox9 is expressed in the different regions of the jaw cartilage structure of developing chickens (Hu et al., 2008), duck, and quail (Eames and Schneider 2008).
- In zebrafish, sox9b has spatiotemporal expression patterns in and around perichondrial cells (Burns et al., 2015), and generally in the lower jaw region of developing sox9b reporter zebrafish (Garcia et al., 2017) (Burns et al., 2016). Sox9 expression has been detected in the dentition of atlantic salmon as well (Huysseune et al., 2008).
- Compared to mice, sox9 expression was identified earlier in the cranial analgen of opposum embryos demonstrating species-specific sox9 expression spatiotemporal patterns (Wakamatsu et al., 2014).
- Few studies have found evidence for relationships between sox9 and molecular signaling pathways that are important for normal development of the craniofacial region. The lines of evidence provide some mechanisms by which sox9 can be involved when disruption of craniofacial cartilage takes place.
- Fertilized chicken eggs infected with retroviruses coding BMP had sox9 expression induced in different regions of the cartilage structure (Hu et al., 2008).
- Exogenous BMP4 added to mouse mandibular explants leads to induction of sox9 expression within 24 hours (Semba et al., 2000). Similarly rat organ cultures exposed to BMP7 for eight days had significantly increased sox9 expression as well as more bone and cartilage, as well as an induction of chondrocyte proliferation and differentiation (Cowan et al., 2006).
- In mouse and chicken in vitro culture systems, addition of BMP induced chondrogenesis, while epidermal growth factor (EGF) repressed both chondrogenesis and also led to sox9 repression (Nonaka et al., 1999).
- Noggin-soaked beads inserted into stage 15 or stage 20 chicken embryos had increased sox9 expression in the maxillary mesenchyme which was associated with ectopic cartilage growth in the stage 15 embryos, and loss of bones in the stage 20 animals (Celá et al., 2016).
- Fibroblast growth factors (FGFs) which play a fundamental role in cartilage formation were added to chicken pluripotent mesenchymal cells which led to both sox repression as well as depression of cartilage matrix production (Bobick et al., 2007). In the case of one rat study, FGF10 electroporated via an expression vector increased both sox9 expression as well as the size of the Meckel’s cartilage (Terao et al., 2011).
- Condylar cartilage explants cultured with a notch signal inhibitor had sox9 expression increased as well as a decrease of proliferation as measured by cyclic B1 expression (Serrano et al., 2014).
- Wnt signaling inhibited by dickkopf-1 in chicken embryos had sox9 expression downregulated, in addition to defects of the maxilla and hypoplasia of the premaxilla and palatine bones (Shimomura et al., 2019).
Empirical Evidence
Empirical evidence and essentiality of KEup for KEdown to occur
- Developing zebrafish exposed to 1ng/mL TCDD had several deformations in the jaw region including in the Meckel’s cartilage which exhibited decreased calcein staining of calcium, and in the craniofacial skeleton which had reduced ossification of the dermal bones. These deformities were associated with a significant reduction of sox9b expression (Burns et al., 2015; Garcia et al., 2017).
- Developing rockfish exposed to 0.5, 5 and 50nM pyrene had decrease of sox9a expression in the craniofacial skeleton region (such as the Meckel’s cartilage, ceratobranchial and pectoral fin blastemas) in a concentration-dependent manner, as well as severe craniofacial deformities starting at 0.5 nM pyrene exposure (Shi et al., 2012).
- In one study, zebrafish injected with sox9b morpholinos had several jaw malformations by 72 hours post fertilization (hpf). Additionally cartilage staining of the jaw showed that the Meckel’s, palatoquadrate, and ceratohyal cartilages were smaller and malformed compared to control morphants (Xiong et al., 2008). A rescue experiment with sox9b mRNA led to 14% of mRNA-injected animals showing normal jaw phenotype.
- Sox9a morpholino knockdown in zebrafish also led to a blockage of cartilage formation (Koskinen et al., 2009).
- In one study, zebrafish exposed to procymidone (0, 10, 100, and 1000 ng/L) significantly repressed sox9 expression (paralog not mentioned) at the 100 and 1000 ng/L concentrations. Significant lower jaw malformations such as, shortened mandibular arch and lower jaw length, were observed at the same concentrations (Wu et al., 2018).
- Investigations into the mutations in the human sox9 coding sequence have identified two novel deletions in the upstream region associated with pierre robin sequence (PRS) (Gordon et al., 2014).
- Sox9 expression purposely ablated in mouse cranial neural crest cells led to an inactivation of sox9 as well as the absence of the condylar cartilage and TMJ malformations (Wang et al., 2011).
- Parathyroid hormone exposure in mice as well as primary chondrocyte cultures from the mandibular condylar cartilage of mice exposed to the hormone had significantly increased sox9 expression as well as a significant increase in cell proliferation in the cartilage and cartilage thickness (Dutra et al., 2021).
- Intermittent hypoxia exposure-induced underdeveloped mandibular ramus and condyles is associated with downregulation of sox9 expression in rats (Lekvijittada et al., 2021). Similarly intermittent nasal obstruction (mouth breathing) in four-week-old rats let to a decrease of sox9 expression in the mesenchymal stem cells of the condylar cartilage, as well as all mandibular parameters being significantly smaller (Wang et al., 2019). However no differences in proliferative ability was seen.
- Mice with altered functional loading had a loss of condylar cartilage, loss of density of mandibular condylar subchondral bone, as well as a decrease in early chondrocyte differentiation, all of which were associated with a decrease in sox9 expression in the jaw region (Chen et al., 2011; Chen et al., 2009). On the other hand, repeated mechanical loading applied to rats had increased sox9 expression as well as a promotion of condylar cartilage growth (Ng et al. 2006).
Uncertainties and Inconsistencies
- Few studies have showed an opposite relationship between sox9 expression and the size of cartilage structures.
- Conditional knockout of setdb1 (histone methyltransferase) specifically in the murine Meckel’s cartilage led to and enlargement of the cartilage structure as well as the proliferation of chondrocytes, however, sox9 expression was significantly repressed (Yahiro et al., 2017).
- Experimental unilateral anterior crossbite created in rats led to decreased ratio of the hypertrophic cartilage layer in the experiment group, which was evidence for obvious cartilage degradation. This was accompanied by induction of sox9 expression (Zhang et al., 2013b).
- One recent zebrafish study using the CRISPR-Cas9 tool, demonstrated that sox9a but not sox9b was required for normal cartilage development (Lin et al., 2021). This is inconsistent with all previous research showing the importance of both sox9a and sox9b for cartilage development in zebrafish.
Known modulating factors
Quantitative Understanding of the Linkage
- Morpholino knockdown of sox9b in developing zebrafish is sufficient to lead to jaw malformations with reduced cartilage structure. Additionally, sox9b heterozygous zebrafish have normal jaw development indicating that there is a threshold for sox9b expression above which jaw can develop normally (Xiong et al., 2008).
Response-response Relationship
Time-scale
Known Feedforward/Feedback loops influencing this KER
Domain of Applicability
- Evidence suggests that sox9 and its function in craniofacial development (and thus its repression leading to craniofacial malformations), is an evolutionarily conserved phenomenon. This relationship appears to exist in almost all vertebrates and invertebrates that encode one or more versions of the sox9 gene.
References
Al-Dujaili M, Milne TJ, Cannon RD, Farella M. 2018. Postnatal expression of chondrogenic and osteogenic regulatory factor mrna in the rat condylar cartilage. Arch Oral Biol. 93:126-132.
Bobick BE, Thornhill TM, Kulyk WM. 2007. Fibroblast growth factors 2, 4, and 8 exert both negative and positive effects on limb, frontonasal, and mandibular chondrogenesis via mek-erk activation. J Cell Physiol. 211(1):233-243.
Burns FR, Lanham KA, Xiong KM, Gooding AJ, Peterson RE, Heideman W. 2016. Analysis of the zebrafish sox9b promoter: Identification of elements that recapitulate organ-specific expression of sox9b. Gene. 578(2):281-289.
Burns FR, Peterson RE, Heideman W. 2015. Dioxin disrupts cranial cartilage and dermal bone development in zebrafish larvae. Aquat Toxicol. 164:52-60.
Celá P, Buchtová M, Veselá I, Fu K, Bogardi JP, Song Y, Barlow A, Buxton P, Medalová J, Francis-West P et al. 2016. Bmp signaling regulates the fate of chondro-osteoprogenitor cells in facial mesenchyme in a stage-specific manner. Dev Dyn. 245(9):947-962.
Chen J, Sobue T, Utreja A, Kalajzic Z, Xu M, Kilts T, Young M, Wadhwa S. 2011. Sex differences in chondrocyte maturation in the mandibular condyle from a decreased occlusal loading model. Calcif Tissue Int. 89(2):123-129.
Chen J, Sorensen KP, Gupta T, Kilts T, Young M, Wadhwa S. 2009. Altered functional loading causes differential effects in the subchondral bone and condylar cartilage in the temporomandibular joint from young mice. Osteoarthritis Cartilage. 17(3):354-361.
Cowan CM, Cheng S, Ting K, Soo C, Walder B, Wu B, Kuroda S, Zhang X. 2006. Nell-1 induced bone formation within the distracted intermaxillary suture. Bone. 38(1):48-58.
Dutra EH, O'Brien MH, Chen PJ, Wei M, Yadav S. 2021. Intermittent parathyroid hormone [1-34] augments chondrogenesis of the mandibular condylar cartilage of the temporomandibular joint. Cartilage. 12(4):475-483.
Eames BF, Schneider RA. 2008. The genesis of cartilage size and shape during development and evolution. Development. 135(23):3947-3958.
Garcia GR, Goodale BC, Wiley MW, La Du JK, Hendrix DA, Tanguay RL. 2017. In vivo characterization of an ahr-dependent long noncoding rna required for proper sox9b expression. Mol Pharmacol. 91(6):609-619.
Gordon CT, Attanasio C, Bhatia S, Benko S, Ansari M, Tan TY, Munnich A, Pennacchio LA, Abadie V, Temple IK et al. 2014. Identification of novel craniofacial regulatory domains located far upstream of sox9 and disrupted in pierre robin sequence. Hum Mutat. 35(8):1011-1020.
Hirouchi H, Kitamura K, Yamamoto M, Odaka K, Matsunaga S, Sakiyama K, Abe S. 2018. Developmental characteristics of secondary cartilage in the mandibular condyle and sphenoid bone in mice. Arch Oral Biol. 89:84-92.
Hu D, Colnot C, Marcucio RS. 2008. Effect of bone morphogenetic protein signaling on development of the jaw skeleton. Dev Dyn. 237(12):3727-3737.
Huang L, Li M, Li H, Yang C, Cai X. 2015. Study of differential properties of fibrochondrocytes and hyaline chondrocytes in growing rabbits. Br J Oral Maxillofac Surg. 53(2):187-193.
Huysseune A, Takle H, Soenens M, Taerwe K, Witten PE. 2008. Unique and shared gene expression patterns in atlantic salmon (salmo salar) tooth development. Dev Genes Evol. 218(8):427-437.
Koskinen J, Karlsson J, Olsson PE. 2009. Sox9a regulation of ff1a in zebrafish (danio rerio) suggests an involvement of ff1a in cartilage development. Comp Biochem Physiol A Mol Integr Physiol. 153(1):39-43.
Lefebvre V, Dvir-Ginzberg M. 2017. Sox9 and the many facets of its regulation in the chondrocyte lineage. Connect Tissue Res. 58(1):2-14.
Lekvijittada K, Hosomichi J, Maeda H, Hong H, Changsiripun C, Kuma YI, Oishi S, Suzuki JI, Yoshida KI, Ono T. 2021. Intermittent hypoxia inhibits mandibular cartilage growth with reduced tgf-β and sox9 expressions in neonatal rats. Sci Rep. 11(1):1140.
Lin Q, He Y, Gui J-F, Mei J. 2021. Sox9a, not sox9b is required for normal cartilage development in zebrafish. Aquaculture and Fisheries. 6(3):254-259.
Ng AF, Yang YO, Wong RW, Hägg EU, Rabie AB. 2006. Factors regulating condylar cartilage growth under repeated load application. Front Biosci. 11:949-954.
Nie X. 2006. Sox9 mrna expression in the developing palate and craniofacial muscles and skeletons. Acta Odontol Scand. 64(2):97-103.
Nonaka K, Shum L, Takahashi I, Takahashi K, Ikura T, Dashner R, Nuckolls GH, Slavkin HC. 1999. Convergence of the bmp and egf signaling pathways on smad1 in the regulation of chondrogenesis. Int J Dev Biol. 43(8):795-807.
Rabie AB, Hägg U. 2002. Factors regulating mandibular condylar growth. Am J Orthod Dentofacial Orthop. 122(4):401-409.
Semba I, Nonaka K, Takahashi I, Takahashi K, Dashner R, Shum L, Nuckolls GH, Slavkin HC. 2000. Positionally-dependent chondrogenesis induced by bmp4 is co-regulated by sox9 and msx2. Dev Dyn. 217(4):401-414.
Serrano MJ, So S, Hinton RJ. 2014. Roles of notch signalling in mandibular condylar cartilage. Arch Oral Biol. 59(7):735-740.
Shi G, Cui Q, Pan Y, Sheng N, Sun S, Guo Y, Dai J. 2017. 6:2 chlorinated polyfluorinated ether sulfonate, a pfos alternative, induces embryotoxicity and disrupts cardiac development in zebrafish embryos. Aquat Toxicol. 185:67-75.
Shi X, He C, Zuo Z, Li R, Chen D, Chen R, Wang C. 2012. Pyrene exposure influences the craniofacial cartilage development of sebastiscus marmoratus embryos. Mar Environ Res. 77:30-34.
Shibata S, Suda N, Suzuki S, Fukuoka H, Yamashita Y. 2006. An in situ hybridization study of runx2, osterix, and sox9 at the onset of condylar cartilage formation in fetal mouse mandible. J Anat. 208(2):169-177.
Shimomura T, Kawakami M, Tatsumi K, Tanaka T, Morita-Takemura S, Kirita T, Wanaka A. 2019. The role of the wnt signaling pathway in upper jaw development of chick embryo. Acta Histochem Cytochem. 52(1):19-26.
Terao F, Takahashi I, Mitani H, Haruyama N, Sasano Y, Suzuki O, Takano-Yamamoto T. 2011. Fibroblast growth factor 10 regulates meckel's cartilage formation during early mandibular morphogenesis in rats. Dev Biol. 350(2):337-347.
Tussellino M, Ronca R, Carotenuto R, Pallotta MM, Furia M, Capriglione T. 2016. Chlorpyrifos exposure affects fgf8, sox9, and bmp4 expression required for cranial neural crest morphogenesis and chondrogenesis in xenopus laevis embryos. Environ Mol Mutagen. 57(8):630-640.
Wakamatsu Y, Nomura T, Osumi N, Suzuki K. 2014. Comparative gene expression analyses reveal heterochrony for sox9 expression in the cranial neural crest during marsupial development. Evol Dev. 16(4):197-206.
Wang X, Sun H, Zhu Y, Tang Y, Xue X, Nie P, Zhu M, Wang B. 2019. Bilateral intermittent nasal obstruction in adolescent rats leads to the growth defects of mandibular condyle. Arch Oral Biol. 106:104473.
Wang Y, Liu C, Rohr J, Liu H, He F, Yu J, Sun C, Li L, Gu S, Chen Y. 2011. Tissue interaction is required for glenoid fossa development during temporomandibular joint formation. Dev Dyn. 240(11):2466-2473.
Watanabe M, Kawasaki K, Kawasaki M, Portaveetus T, Oommen S, Blackburn J, Nagai T, Kitamura A, Nishikawa A, Kodama Y et al. 2016. Spatio-temporal expression of sox genes in murine palatogenesis. Gene Expr Patterns. 21(2):111-118.
Wu Y, Zuo Z, Chen M, Zhou Y, Yang Q, Zhuang S, Wang C. 2018. The developmental effects of low-level procymidone towards zebrafish embryos and involved mechanism. Chemosphere. 193:928-935.
Xiong KM, Peterson RE, Heideman W. 2008. Aryl hydrocarbon receptor-mediated down-regulation of sox9b causes jaw malformation in zebrafish embryos. Mol Pharmacol. 74(6):1544-1553.
Yahiro K, Higashihori N, Moriyama K. 2017. Histone methyltransferase setdb1 is indispensable for meckel's cartilage development. Biochem Biophys Res Commun. 482(4):883-888.
Zhang H, Zhao X, Zhang Z, Chen W, Zhang X. 2013a. An immunohistochemistry study of sox9, runx2, and osterix expression in the mandibular cartilages of newborn mouse. Biomed Res Int. 2013:265380.
Zhang X, Dai J, Lu L, Zhang J, Zhang M, Wang Y, Guo M, Wang X, Wang M. 2013b. Experimentally created unilateral anterior crossbite induces a degenerative ossification phenotype in mandibular condyle of growing sprague-dawley rats. J Oral Rehabil. 40(7):500-508.