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In humans, OFD1 is mutated in oral-facial-digital type I syndrome leading to prenatal death in hemizygous males and dysmorphic faces and
brain malformations, with polycystic kidneys presenting later in life in heterozygous females. To elucidate the function of
Ofd1, we have studied its function during zebrafish embryonic development. In wild-type e...
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Context 1
... address the requirements for Ofd1 during development, we injected ofd1 MOs into one-cell stage embryos, using either a translation blocking (ATG) or one of two splicing-perturbing (SPL6 and SPL7) MOs. Each produced a similar dysmorphic spectrum (Table 1 and data not shown). By 28 hpf, ofd1 MO-injected embryos displayed upwards or downwards body axis curvatures ( Fig. 2A and B), with aberrant curvature maintained over the following days (Fig. 2C-H and data not shown). ...
Context 2
... also measured gene expression in mid-gastrulation embryos using microarrays. Table 3 lists genes up-or down-regulated .2-fold in ofd1 MO-injected embryos (see Supplementary Material, Table S1 for the complete set) and we verified changed expression of selected genes by real-time RT -PCR (Fig. 7). Strikingly, in the microarray study, the greatest-fold upregulated transcript was ofd1 itself. ...
Citations
... Laterality was randomized in the brain, heart, and viscera, which may have been caused by short cilia with disrupted axonemes and perturbed intravesicular fluid flow in Kupffer vesicles. The author concluded that OFD1 is necessary for ciliary movement and function in zebrafish, which supports the data suggesting that OFD1 is essential for ciliary function in mice [35]. ...
Background
Left–right laterality disorders are a heterogeneous group of disorders caused by an altered position or orientation of the thoracic and intra-abdominal organs and vasculature across the left–right axis. They mainly include situs inversus and heterotaxy. Those disorders are complicated by cardiovascular abnormalities significantly more frequently than situs solitus.
Methods
In this study, 16 patients with a fetal diagnosis of laterality disorder with congenital heart defects (CHD) were evaluated with a single nucleotide polymorphism array (SNP-arry) combined with whole-exome sequencing (WES).
Results
Although the diagnostic rate of copy number variations was 0 and the diagnostic rate of WES was 6.3% (1/16), the likely pathogenic gene DNAH11 and the candidate gene OFD1 were ultimately identified. In addition, novel compound heterozygous mutations in the DNAH11 gene and novel hemizygous variants in the OFD1 gene were found. Among the combined CHD, a single atrium/single ventricle had the highest incidence (50%, 8/16), followed by atrioventricular septal defects (37.5%, 6/16). Notably, two rare cases of common pulmonary vein atresia (CPVA) were also found on autopsy.
Conclusion
This study identified the types of CHD with a high incidence in patients with laterality disorders. It is clear that WES is an effective tool for diagnosing laterality disorders and can play an important role in future research.
... Approximately 30% of all ciliopathies can be classified by their craniofacial phenotypes, including cleft lip/palate, craniosynostosis, and micrognathia (Schock and Brugmann, 2017). Human patients and animal models that present with craniofacial ciliopathies have an array of mandibular phenotypes such as micrognathia, dysmorphic skeletal elements, and reduced bone density (Adel Al-Lami et al., 2016;Bonatto Paese et al., 2021a;Cela et al., 2018;Ferrante et al., 2009;Kantaputra et al., 2023;Kitamura et al., 2020;Kolpakova-Hart et al., 2007;Zhang et al., 2011). While significant progress has been made in understanding the molecular etiologies of craniofacial ciliopathies, comprehensive knowledge of how individual ciliary proteins regulate the spatial patterns of early molecular signals that contribute to musculoskeletal tissue patterning is currently lacking. ...
The mandible is composed of several musculoskeletal tissues including bone, cartilage, and tendon that require precise patterning to ensure structural and functional integrity. Interestingly, most of these tissues are derived from one multipotent cell population called cranial neural crest cells (CNCCs). How CNCCs are properly instructed to differentiate into various tissue types remains nebulous. To better understand the mechanisms necessary for the patterning of mandibular musculoskeletal tissues we utilized the avian mutant talpid2 (ta2) which presents with several malformations of the facial skeleton including dysplastic tendons, mispatterned musculature, and bilateral ectopic cartilaginous processes extending off Meckel's cartilage. We found an ectopic epithelial BMP signaling domain in the ta2 mandibular prominence (MNP) that correlated with the subsequent expansion of SOX9+ cartilage precursors. These findings were validated with conditional murine models suggesting an evolutionarily conserved mechanism for CNCC-derived musculoskeletal patterning. Collectively, these data support a model in which cilia are required to define epithelial signal centers essential for proper musculoskeletal patterning of CNCC-derived mesenchyme.
... Multicilia are mainly found in the olfactory organ, pronephric/ kidney duct, ependymal cells and fallopian tubes, while motile cilia are primarily distributed in the early otic vesicle, KV, brain ventricles, central canal, pronephric/kidney duct, fallopian tubes and periphery of olfactory organs. In addition to these types of cilia, there are immotile multi-cilia in olfactory neurons as well as single immotile cilia in KV and other tissues (figures 2 and 3) [37,[73][74][75][76]. Most motile cilia in zebrafish exhibit an ultrastructure of '9 + 2', but there are '9 + 2' or '9 + 0' motile cilia in the neural tube and KV [37,76]. ...
... In addition to these types of cilia, there are immotile multi-cilia in olfactory neurons as well as single immotile cilia in KV and other tissues (figures 2 and 3) [37,[73][74][75][76]. Most motile cilia in zebrafish exhibit an ultrastructure of '9 + 2', but there are '9 + 2' or '9 + 0' motile cilia in the neural tube and KV [37,76]. Most primary cilia exhibit an ultrastructure of '9 + 0', while the kinocilia in the otic vesicle and cilia in the olfactory neuron are '9 + 2' immotile cilia [36,[77][78][79]. ...
Cilia are hair-like organelles that protrude from the surface of eukaryotic cells and are present on the surface of nearly all human cells. Cilia play a crucial role in signal transduction, organ development and tissue homeostasis. Abnormalities in the structure and function of cilia can lead to a group of human diseases known as ciliopathies. Currently, zebrafish serves as an ideal model for studying ciliary function and ciliopathies due to its relatively conserved structure and function of cilia compared to humans. In this review, we will summarize the different types of cilia that present in embryonic and adult zebrafish, and provide an overview of the advantages of using zebrafish as a vertebrate model for cilia research. We will specifically focus on the roles of cilia during zebrafish organogenesis based on recent studies. Additionally, we will highlight future prospects for ciliary research in zebrafish.
... Further research revealed that deletion of BBS8 disrupted asymmetric accumulation of the core PCP protein Vangl2 in cochlear cells, suggesting a role for BBS8 possibly upstream of core PCP asymmetry. Loss of INVERSIN and OFD1 led to PCP-regulated convergent extension defects in vertebrates also supported the involvement of cilium in non-canonical WNT signaling [61,71]. Besides that, the deletion of BBSome had increased the release of small EVs (smEVs) loaded with WNT-related molecules and smEVs derived from ciliopathy patient renal tissues dampened the WNT response in target cells, in contrast with control tissues [72]. ...
Epithelial-to-mesenchymal transition (EMT), which plays an essential role in development, tissue repair and fibrosis, and cancer progression, is a reversible cellular program that converts epithelial cells to mesenchymal cell states characterized by motility-invasive properties. The mostly signaling pathways that initiated and controlled the EMT program are regulated by a solitary, non-motile organelle named primary cilium. Acting as a signaling nexus, primary cilium dynamically concentrates signaling molecules to respond to extracellular cues. Recent research has provided direct evidence of connection between EMT and primary ciliogenesis in multiple contexts, but the mechanistic understanding of this relationship is complicated and still undergoing. In this review, we describe the current knowledge about the ciliary signaling pathways involved in EMT and list the direct evidence that shows the link between them, trying to figure out the intricate relationship between EMT and primary ciliogenesis, which may aid the future development of primary cilium as a novel therapeutic approach targeted to EMT.
... have been implicated in primary cilia biogenesis and function. The phenotypic spectrum of OFD syndromes can overlap with other known ciliopathies, and mutations in OFD genes can also cause other types of ciliopathies [16][17][18][19][20][21][22] . Of the 12 known genes involved in OFDs, the majority encode structural components of the primary cilium, such as OFD1 23 and TMEM231 24 . ...
Oral-facial-digital (OFD) syndromes are a heterogeneous group of congenital disorders characterized by malformations of the face and oral cavity, and digit anomalies. Mutations within 12 cilia-related genes have been identified that cause several types of OFD, suggesting that OFDs constitute a subgroup of developmental ciliopathies. Through homozygosity mapping and exome sequencing of two families with variable OFD type 2, we identified distinct germline variants in INTS13, a subunit of the Integrator complex. This multiprotein complex associates with RNA Polymerase II and cleaves nascent RNA to modulate gene expression. We determined that INTS13 utilizes its C-terminus to bind the Integrator cleavage module, which is disrupted by the identified germline variants p.S652L and p.K668Nfs*9. Depletion of INTS13 disrupts ciliogenesis in human cultured cells and causes dysregulation of a broad collection of ciliary genes. Accordingly, its knockdown in Xenopus embryos leads to motile cilia anomalies. Altogether, we show that mutations in INTS13 cause an autosomal recessive ciliopathy, which reveals key interactions between components of the Integrator complex.
... Mutations in human POLR1A gene that cause the Cincinnati type of acrofacial dysostosis and in OFD1 gene that induce the orofaciodigital syndrome type I have also been modified to obtain models of diseases in group 34 (Mortier et al., 2019, Supplementary Table 1). Following mutations or downregulations of polr1a (Weaver et al., 2015;Watt et al., 2018) and ofd1 (Ferrante et al., 2009) genes, phenotypes resembling human symptoms are obtained. These phenotypes are lethality, craniofacial disorders, small eyes, otic vesicles, bent pectoral fins, small body sizes, and abnormal hearts or notochords. ...
In 2019, the Nosology Committee of the International Skeletal Dysplasia Society provided an updated version of the Nosology and Classification of Genetic Skeletal Disorders. This is a reference list of recognized diseases in humans and their causal genes published to help clinician diagnosis and scientific research advances. Complementary to mammalian models, zebrafish has emerged as an interesting species to evaluate chemical treatments against these human skeletal disorders. Due to its versatility and the low cost of experiments, more than 80 models are currently available. In this article, we review the state-of-art of this “aquarium to bedside” approach describing the models according to the list provided by the Nosology Committee. With this, we intend to stimulate research in the appropriate direction to efficiently meet the actual needs of clinicians under the scope of the Nosology Committee.
... The cilia were severely disrupted or lost, producing defects in laterality and Hh-dependent tissue patterning [318,326]. The zebrafish ofd1 morphants also displayed laterality defects, due to cilia abnormalities in the Kupffer´s vesicle, as well as additional ciliopathy features [327]. These data suggest that the decreased levels of endogenous and centrosome-competent OFD1 in the FGFR2-OFD1 cancers may lead to deregulated ciliogenesis and cilia signaling, potentially contributing to neoplastic transformation. ...
A single primary cilium projects from most vertebrate cells to guide cell fate decisions. A growing list of signaling molecules is found to function through cilia and control ciliogenesis, including the fibroblast growth factor receptors (FGFR). Aberrant FGFR activity produces abnormal cilia with deregulated signaling, which contributes to pathogenesis of the FGFR-mediated genetic disorders. FGFR lesions are also found in cancer, raising a possibility of cilia involvement in the neoplastic transformation and tumor progression. Here, we focus on FGFR gene fusions, and discuss the possible mechanisms by which they function as oncogenic drivers. We show that a substantial portion of the FGFR fusion partners are proteins associated with the centrosome cycle, including organization of the mitotic spindle and ciliogenesis. The functions of centrosome proteins are often lost with the gene fusion, leading to haploinsufficiency that induces cilia loss and deregulated cell division. We speculate that this complements the ectopic FGFR activity and drives the FGFR fusion cancers.
... This ciliary phenotype was recapitulated in Medaka fish where knock-down of TBC1D31 markedly altered ciliogenesis, cilium morphology and development. Importantly, this phenotype might be a consequence of lack of OFD1, because previous studies have shown that depletion of OFD1 directly induces this phenotype in zebrafish (Ferrante et al, 2009;Liu et al, 2014). However, we found that the concomitant knockdown of TBC1D31 and overexpression of wild-type OFD1 was insufficient to restore normal cilia size and embryogenesis (Fig 7A-C), possibly because TBC1D31 depletion directly affects praja2/PKA/ OFD1 molecular network at centrosome. ...
The primary cilium is a microtubule-based sensory organelle that dynamically links signalling pathways to cell differentiation, growth, and development. Genetic defects of primary cilia are responsible for genetic disorders known as ciliopathies. Orofacial digital type I syndrome (OFDI) is an X-linked congenital ciliopathy caused by mutations in the OFD1 gene and characterized by malformations of the face, oral cavity, digits and, in the majority of cases, polycystic kidney disease. OFD1 plays a key role in cilium biogenesis. However, the impact of signalling pathways and the role of the ubiquitin-proteasome system (UPS) in the control of OFD1 stability remain unknown. Here, we identify a novel complex assembled at centrosomes by TBC1D31, including the E3 ubiquitin ligase praja2, protein kinase A (PKA), and OFD1. We show that TBC1D31 is essential for ciliogenesis. Mechanistically, upon G-protein-coupled receptor (GPCR)-cAMP stimulation, PKA phospho-rylates OFD1 at ser735, thus promoting OFD1 proteolysis through the praja2-UPS circuitry. This pathway is essential for ciliogenesis. In addition, a non-phosphorylatable OFD1 mutant dramatically affects cilium morphology and dynamics. Consistent with a role of the TBC1D31/praja2/OFD1 axis in ciliogenesis, alteration of this molecular network impairs ciliogenesis in vivo in Medaka fish, resulting in developmental defects. Our findings reveal a multi-functional transduction unit at the centrosome that links GPCR signalling to ubiquitylation and proteolysis of the ciliopathy protein OFD1, with important implications on cilium biology and development. Derangement of this control mechanism may underpin human genetic disorders.
... OFD1 is a centrosomal protein, and mutations in the coding region of the gene are associated with Xlinked Joubert syndrome (JBTS10), Simpson-Golabi-Behmel syndrome type 2 (SGBS2) and orofaciodigital syndrome, a ciliopathy (35)(36)(37). Importantly, just like WDR8, OFD1 also has functions both in the stabilization of microtubules during cell division and in the formation of primary cilium (35,38,39). Thus, the substitution of proline to leucine may alter the WD dipeptidemediated interaction with the binding partners that are involved in cell division. ...
Isolated Microspherophakia (MSP) is an autosomal recessive disorder characterised by a smaller than normal spherical lens. Till date, LTBP2 is the only gene shown to cause MSP. We used homozygosity mapping and whole-exome sequencing and identified a homozygous mutation, c.1148C > T (p.Pro383Leu), in the WDR8 (or WRAP73) gene in two Indian MSP families. In vitro experiments showed that the missense mutation renders the protein unstable. WDR8 is a centriolar protein that has important roles in centrosomal assembly, spindle pole formation, and ciliogenesis. Co-immunoprecipitation experiments from HeLa cells indicated that the mutation interferes with the interaction of WDR8 with its binding partners. In zebrafish, both morpholino-mediated knockdown and CRISPR/Cas knockout of wdr8 resulted in decreased eye and lens size. The lack of wdr8 affected cell cycle progression in the retinal cells, causing a reduction in cell numbers in the retina and lens. The reduction in eye size and the cell cycle defects were rescued by exogenous expression of the human wild type WDR8. However, the human mutant WDR8 (p.Pro383Leu) was unable to rescue the eye defects, indicating that the missense mutation abrogates WDR8 protein function. Thus, our zebrafish results suggested that WDR8 is the causative gene for MSP in these Indian families.
... The ofd1 mutant zebrafish (Danio Rerio) model also helped to elucidate the role of OFD1 in the regulation of cilia function [49]. This zebrafish mutant produced a typical ciliary phenotype [50] including curved body and laterality defects. ...
... Furthermore OFD1, as part of the TIP60 complex, could act in the nucleus to regulate the transcriptional program that directs the development of tissues and organs affected in OFDI syndrome. The observation that down-regulation of ofd1 in zebrafish leads to significant alteration in levels of genes transcription supports this hypothesis [49]. ...
The OFD1 gene was initially identified as the gene responsible for the X-linked dominant male lethal OFD type I syndrome, a developmental disorder ascribed to cilia disfunction. The transcript has been subsequently associated to four different X-linked recessive conditions, namely Joubert syndrome, retinitis pigmentosa, primary ciliary dyskinesia and Simpson-Golabi-Behmel type 2 syndrome. The centrosomal/basal body OFD1 protein has indeed been shown to be required for primary cilia formation and left-right asymmetry. The protein is also involved in other tasks, e.g. regulation of cellular protein content, constrain of the centriolar length, chromatin remodeling at DNA double strand breaks, control of protein quality balance and cell cycle progression, which might be mediated by non-ciliary activities. OFD1 represents a paradigmatic model of a protein that performs its diverse actions according to the cell needs and depending on the subcellular localization, the cell type/tissue and other possible factors still to be determined. An increased number of multitask protein, such as OFD1, may represent a partial explanation to human complexity, as compared with less complex organisms with an equal or slightly lower number of proteins.
