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GDF5 W414R shows impaired Bmpr1a signaling in a SBE-Luciferase reporter gene assay. NIH/3T3 cells were transfected with the BMP type I receptors, Bmpr1a or Bmpr1b, as well as with wild type GDF5 and the GDF5 variants GDF5 W414R , GDF5 R399C and GDF5 E491K . As reporter, the SMAD binding element (SBE) was used and firely luciferase was normalized against TK-Renilla luciferase. A : No Bmp type I receptor was co-expressed which resulted in a weak SBE reporter activation for wild type GDF5 and GDF5 E491K , whereas in case of GDF5 W414R and GDF5 R399C signaling activity was absent. B : Bmpr1a co-expression increased the signaling activity of wild type GDF5 and GDF5 E491K ; however, GDF5 W414R and GDF5 R399C were not able to induce reporter gene expression. C : Co-expression of Bmpr1b further increased the signaling activity of wild type GDF5 and GDF5 E491K compared to co-expression with Bmpr1a . In case of GDF5 W414R and GDF5 R399C , Bmpr1b co-expression rescued their signaling activity. The means of triplicate measurements are shown, error bars indicate standard deviation and a represent experiment is shown. Statistical analysis was performed using a two-tailed Student’s t test (n.s.: not significant; *p # 0.05; **p # 0.01). Significances are related to the respective wild type GDF5 value. doi:10.1371/journal.pgen.1003846.g004
Source publication
Growth and Differentiation Factor 5 (GDF5) is a secreted growth factor that belongs to the Bone Morphogenetic Protein (BMP) family and plays a pivotal role during limb development. GDF5 is a susceptibility gene for osteoarthritis (OA) and mutations in GDF5 are associated with a wide variety of skeletal malformations ranging from complex syndromes s...
Citations
... 13 A variant of GDF5 has been shown to be related to BDA1 (OMIM #112500), BDA2 (OMIM #112600), and BDC (OMIM #113100). 6,14,15 Homozygous deletion variants in GDF5 lead to the development of extremely short fingers and limbs, which is associated with various types of acrochondrodysplasia. 16,17 In contrast to loss-of-function GDF5 variants, gain-of-function GDF5 variants increase the chondrogenous activity and are associated with SYM1 and SYNS2. 6,17 SYM1 is characterized by proximal interphalangeal stiffness, in which the carpus and tarsus are fused. ...
... The diagnosis and classification of BDA1 and SYNS2 complied with the latest guidelines. 16 All the participants signed informed consent for this study, which was approved by the Ethics Committee of the Central Hospital of Wuhan. ...
Objective
This study aimed to identify the molecular defects and clinical manifestations in a Chinese family with brachydactyly (BD) type A1 (BDA1) and multiple‐synostoses syndrome 2 (SYNS2).
Methods
A Chinese family with BDA1 and SYNS2 was enrolled in this study. Whole‐exome sequencing was used to analyze the gene variants in the proband. The sequences of the candidate pathogenic variant in GDF5 was validated via Sanger sequencing. I‐TASSER and PyMOL were used to analyze the functional domains of the corresponding mutant proteins.
Results
The family was found to have an autosomal‐dominantly inherited combination of BDA1 and SYNS2 caused by the S475N variant in the GDF5 gene. The variant was located within the functional region, and the mutated residue was found to be highly conserved among species. Via bioinformatic analyses, we predicted this variant to be deleterious, which perturb the protein function. The substitution of the negatively charged amino acid S475 with the neutral N475 was predicted to disrupt the formation of salt bridges with Y487 and impair the structure, stability, and function of the protein, consequently, the abnormalities in cartilage and bone development ensue.
Conclusions
A single genetic variant (S475N) which disrupt the formation of salt bridges with Y487, in the interface of the antagonist‐ and receptor‐binding sites of GDF5 concurrently causes two pathological mechanisms. This is the first report of this variant, identified in a Chinese family with BDA1 and SYNS2.
... GDF5 belongs to the family of bone morphogenetic proteins (BMP), which is involved in bone growth and repair, such as the proliferation, differentiation, angiogenesis, and bone and cartilage formation. Research shows that GDF5 genetic polymorphisms are closely related to OA [46,47]. However, it is not completely clear what role GDF5 plays in the occurrence and development of OA, and how it affects other signal pathways. ...
Background
Osteoarthritis (OA) is caused by a complex set of pathophysiological factors. The genetic factors involved in the occurrence and progress of the disease have been widely discussed by scholars. It was found that growth differentiation factor 5 (GDF5) gene polymorphisms may be linked to OA susceptibility, which has been controversial and needs to be further confirmed by an updated meta-analysis.
Objectives
We examined the association between GDF5 rs143383 single nucleotide polymorphism (SNP) and OA susceptibility.
Methods
All relevant articles that met the criteria are retrieved and included, and the search deadline is June 2022. The allele frequencies and different genotype frequencies of GDF5 rs143383 loci in each study were extracted and statistically analyzed by R4.1.3 software, and the different genetic models were analyzed based on their odds ratio (OR) and 95% confidence interval (CI).
Results
The meta-analysis explained that GDF5 rs143383 SNP was crucial correlated with OA in all patients with OA of knee, hip and hand. The codominant gene model in the whole crowd (OR = 1.17, 95% CI 1.07–1.27, P < 0.01) enlightened that OA was vitally associated with GDF5 gene polymorphism. At the same time, we did a subgroup analysis based on ethnicity. The codominant gene model (OR = 1.31, 95% CI 1.12–1.53, P < 0.01) in Asian population, the codominant homozygote model (OR = 1.28, 95% CI 1.14–1.43), codominant heterozygote gene model (OR = 1.12, 95% CI 1.01–1.23, P = 0.02), and dominant gene model (OR = 1.19, 95% CI 1.09–1.31, P < 0.01) in Caucasian are analyzed by subgroup analysis. It means that there is a momentous relationship between the GDF5rs143383 gene polymorphism and OA, especially among Caucasians. In addition, we also discussed different types of OA separately and discover that the GDF5rs143383 gene polymorphism was relevant for knee osteoarthritis (KOA) and hand osteoarthritis, and it was more significant in the Caucasian population. But due to the high heterogeneity in hip osteoarthritis, it could not be accurately concluded. Furthermore, we also analyzed the osteoarthritis of different genders and found that the GDF5 rs143383 SNP was associated with both men and women and was still significant in the Caucasian population.
Conclusion
We found a close association between osteoarthritis and GDF5rs143383SNP in this study. From the analysis of each group, we got the same conclusion in KOA and hand OA, but which need further verification in hip OA. Considering gender, we found a close relationship between GDF5 rs143383 SNP and OA of the knee, hip and hand, both for men and women. This conclusion is more obvious in Caucasian people.
... 6,7 Cualquier alteración molecular en dicho gen podría provocar condrólisis y otras consecuencias en el cartílago, el hueso y el desarrollo articular. 6,19 Sin embargo, no solo los factores genéticos están relacionados con la DCC y su predisposición a OA. Como hemos observado en el presente artículo, factores inflamatorios e inmunológicos desencadenan un proceso, originado quizás por la inestabilidad mecánica producida por la DCC, que causa inflamación crónica de bajo grado en todos los componentes articulares, la cual se manifiesta clínicamente como fibrosis, degradación del cartílago articular, engrosamiento del hueso subcondral, formación de osteofitos, sinovitis, degradación de meniscos y ligamentos, e hipertrofia capsular. ...
... ,19,20 Figura 3. Gen GDF5 y su participación en un interactoma por coexpresión, vecindad o genes fusionados. Fuente: adaptado de STRING Database.20 ...
... Fuente: adaptado de Exploring Data.21 Rev Col Or Tra | https://doi.org/10.58814/01208845.19 ...
Introducción. La displasia del desarrollo de la cadera (DDC) abarca un conjunto de anormalidades relacionadas con el proceso de maduración del acetábulo y del tercio proximal del fémur Si no se trata de manera adecuada y oportuna, los pacientes con esta condición pueden desarrollar osteoartritis (OA) eventualmente. Objetivo. Recopilar y sintetizar evidencia científica publicada entre enero de 2000 y febrero de 2023 sobre la fisiopatología de la DDC y su relación con el desarrollo de OA de cadera en términos de los mecanismos fisiopatológicos genéticos, inflamatorios e inmunológicos. Materiales y métodos. Se realizó una revisión de la literatura en bases de datos de literatura biomédica (PubMed/Medline, Embase, SciELO) y herramientas bioinformáticas (e-Ensambl, STRING), mediante términos como “displasia de cadera”, “osteoartritis”, “etiología” y “genes”. Se incluyeron estudios observacionales clínicos y genéticos realizados en humanos. Resultados. La búsqueda inicial arrojó 349 registros, de los cuales 23 cumplieron los criterios de elegibilidad. Los genes que interactúan con módulos genéticos parecen participar en el desarrollo articular y la etiología de las enfermedades relacionadas con el cartílago y el hueso; sin embargo, la inestabilidad mecánica producida por la DCC activa factores inflamatorios e inmunológicos, predisponiendo OA. A partir de la información encontrada, se puede considerar que existe una relación muy estrecha entre DDC y OA. Conclusiones. Conocer los mecanismos fisiopatológicos genéticos, inflamatorios e inmunológicos de DDC y OA favorece la realización de un diagnóstico oportuno y, en consecuencia, posibilita brindar un tratamiento adecuado para disminuir y controlar el daño a largo plazo y mejorar la calidad de vida del paciente.
... Examining chondrogenic cell states, we observed that although chondrogenesis appear to proceed overall normally in the mutant, the reduction in distal FGF signaling led to a reduced diversity of spatially distributed chondrogenic cell states: We found fewer distal Sox9-positive cell states expressing Eomes [base of digit 4 (60)] and Irx1 [tip of digits 2 to 4 (61)] (Fig. 5, C and D). These distal chondrogenic cell states have been proposed to act as regulation centers expressing different BMP pathway components involved in joint formation, including Gdf5, Inhb, Nog, and Chrdl1 (62)(63)(64)(65). Although the depletion of these cell states is only partial, no expression of these signaling-related genes was detected in the mutant data ( fig. ...
The coordinated differentiation of progenitor cells into specialized cell types and their spatial organization into distinct domains is central to embryogenesis. Here, we developed and applied an unbiased spatially resolved single-cell transcriptomics method to identify the genetic programs underlying the emergence of specialized cell types during mouse limb development and their spatial integration. We identify multiple transcription factors whose expression patterns are predominantly associated with cell type specification or spatial position, suggesting two parallel yet highly interconnected regulatory systems. We demonstrate that the embryonic limb undergoes a complex multiscale reorganization upon perturbation of one of its spatial organizing centers, including the loss of specific cell populations, alterations of preexisting cell states’ molecular identities, and changes in their relative spatial distribution. Our study shows how multidimensional single-cell, spatially resolved molecular atlases can allow the deconvolution of spatial identity and cell fate and reveal the interconnected genetic networks that regulate organogenesis and its reorganization upon genetic alterations.
... Functional single-nucleotide polymorphisms linked to reduced Gdf5 gene transcription are currently the most consistent risk factor for adult-onset OA across human populations 16,17 . Interestingly, skeletal defects resulted from dysfunction of miR-17~92 cluster showed shortening of phalangeal elements and bony fusions of the joints 12 , which are also observed with GDF5 deficiency 18 , suggesting an inter-connection between GDF5 and miR- 17. In this study, we used destabilization of the medial meniscus (DMM)-induced mouse OA model, as well as a series of gain-and loss-of-function analyses plus single-cell RNA sequencing (scRNAseq) to examine the regulation and function of miR-17, together with molecular mechanisms underlying OA pathophysiology. ...
Damaged hyaline cartilage has no capacity for self-healing, making osteoarthritis (OA) “difficult-to-treat”. Cartilage destruction is central to OA patho-etiology and is mediated by matrix degrading enzymes. Here we report decreased expression of miR-17 in osteoarthritic chondrocytes and its deficiency contributes to OA progression. Supplementation of exogenous miR-17 or its endogenous induction by growth differentiation factor 5, effectively prevented OA by simultaneously targeting pathological catabolic factors including matrix metallopeptidase-3/13 (MMP3/13), aggrecanase-2 (ADAMTS5), and nitric oxide synthase-2 (NOS2). Single-cell RNA sequencing of hyaline cartilage revealed two distinct superficial chondrocyte populations (C1/C2). C1 expressed physiological catabolic factors including MMP2, and C2 carries synovial features, together with C3 in the middle zone. MiR-17 is highly expressed in both superficial and middle chondrocytes under physiological conditions, and maintains the physiological catabolic and anabolic balance potentially by restricting HIF-1α signaling. Together, this study identified dual functions of miR-17 in maintaining cartilage homeostasis and prevention of OA. Osteoarthritic (OA) is characterized by progressive destruction of joint cartilage. Here, the authors show that microRNA-17 plays a dual role in maintaining cartilage homeostasis and in the prevention of osteoarthritis, by targeting hypoxia-inducible factor-1α as well as multiple matrix-degrading enzymes.
... The dominant negative effect of a missense mutation in GDF5 in a family with Grebe type chondrodysplasia was shown to be caused by the inability to secrete other related BMP family members (Thomas et al., 1997). Gain-of-function pathogenic variants in GDF5 are associated with type 2 multiple synostoses syndrome and proximal symphalangism (Degenkolbe et al., 2013;Plett et al., 2008). It is possible that duplication of this gene alters GDF5 signaling and contributes to the shorthands and feet seen in our and other patients. ...
Background
Reports of interstitial duplication of chromosome 20q11 are rare with only nine published patients to date.
Methods
We performed karyotype and chromosomal microarray analysis on a peripheral blood sample for our patient and reviewed the genes in the region to provide genotype–phenotype correlation.
Results
Clinical features of the patient include minor dysmorphic facial features, shorthands and feet, bilateral conductive hearing loss, global developmental delay, and behavioral issues with attention deficit hyperactivity disorder. Together with previously published cases of 20q11 duplication, we show that patients with overlapping duplications share a similar clinical phenotype of dysmorphic craniofacial features and developmental delay.
Conclusion
We report an 8-year-old girl with a 9.1 Mb interstitial duplication of chromosome 20q11.22q13.11. Our observations suggest that a novel duplication syndrome and documentation of similar cases will further help clarify the phenotype.
... 77,78 In vitro, Gdf5 promotes chondrogenesis, but reduces Bmp2-mediated chondrogenesis in the absence of Bmpr1b. 78,79 Similarly, in vivo, Gdf5 promotes chondrogenesis prior to interzone formation or when applied ectopically, 80 but Gdf5 expression is associated with reduced pSmad1/5 activity in the interzone. 81 Bmpr1a is expressed in interzone cells and Bmpr1b is only expressed in the nearby periarticular and epiphyseal cells, 82,83 suggesting that the availability of Type I receptors is a critical component of Gdf5 function (for detailed review, see Lyons and Rosen 37 ). ...
... Human inactivating mutations in GDF5 lead to various forms of brachydactyly which may include loss of specific medial phalanges. 79,84 As discussed above, the murine limb phenotype caused by Gdf5 inactivation is very similar to that caused by Acvr1 R206H expression. This suggests either that a primary function of Gdf5 in joint development is to directly inhibit wild-type Acvr1, or that Gdf5 functions generally to inhibit BMP pathway activity in the interzone and that dysregulated signaling through Acvr1 R206H escapes or makes negligible this inhibitory function. ...
Fibrodysplasia ossificans progressiva (FOP) is an ultra‐rare genetic disease caused by increased BMP pathway signaling due to mutation of ACVR1, a bone morphogenetic protein (BMP) type 1 receptor. The primary clinical manifestation of FOP is extra‐skeletal bone formation (heterotopic ossification) within soft connective tissues. However, the underlying ACVR1 mutation additionally alters skeletal bone development and nearly all people born with FOP have bilateral malformation of the great toes as well as other skeletal malformations at diverse anatomic sites. The specific mechanisms through which ACVR1 mutations and altered BMP pathway signaling in FOP influence skeletal bone formation during development remain to be elucidated; however, recent investigations are providing a clearer understanding of the molecular and developmental processes associated with ACVR1‐regulated skeletal formation.
... 35 Other genes where a variant is reported to cause both GoF and LoF effects are RET, FLNA, GDF5 and SMCHD1. [36][37][38][39] In general, multinucleotide variants are potentially more damaging than single-nucleotide variants, illustrated by our family. 40 The most intriguing finding was that the delins also affected a hitherto not confirmed transcript using a non-canonical reading frame-CDKN1C-201-which represented ~40% of all CDKN1C transcripts in the family (figure 5A). ...
Background
Loss-of-function mutations in CDKN1C cause overgrowth, that is, Beckwith-Wiedemann syndrome (BWS), while gain-of-function variants in the gene’s PCNA binding motif cause a growth-restricted condition called IMAGe syndrome. We report on a boy with a remarkable mixture of both syndromes, with developmental delay and microcephaly as additional features.
Methods
Whole-exome DNA sequencing and ultra-deep RNA sequencing of leucocyte-derived and fibroblast-derived mRNA were performed in the family.
Results
We found a maternally inherited variant in the IMAGe hotspot region: NM_000076.2( CDKN1C ) c.822_826delinsGAGCTG. The asymptomatic mother had inherited this variant from her mosaic father with mild BWS features. This delins caused tissue-specific frameshifting resulting in at least three novel mRNA transcripts in the boy. First, a splice product causing CDKN1C truncation was the likely cause of BWS. Second, an alternative splice product in fibroblasts encoded IMAGe-associated amino acid substitutions. Third, we speculate that developmental delay is caused by a change in the alternative CDKN1C-201 (ENST00000380725.1) transcript, encoding a novel isoform we call D (UniProtKB: A6NK88). Isoform D is distinguished from isoforms A and B by alternative splicing within exon 1 that changes the reading frame of the last coding exon. Remarkably, this delins changed the reading frame back to the isoform A/B type, resulting in a hybrid D–A/B isoform.
Conclusion
Three different cell-type-dependent RNA products can explain the co-occurrence of both BWS and IMAGe features in the boy. Possibly, brain expression of hybrid isoform D–A/B is the cause of developmental delay and microcephaly, a phenotypic feature not previously reported in CDKN1C patients.
... Distinct aspects of Gdf5 expression have been reported in several publications that combined have shown Gdf5 expression between E11.5 and E14.5 in the presumptive sites of joint development and the forming joint interzone ( Chen et al. 2016;Degenkolbe et al. 2013;Hellman et al. 2012;Houweling et al. 2001;Huang et al. 2016;Meech et al. 2005;Ota et al. 2007;Perez et al. 2010;Seemann et al. 2005;Sohaskey et al. 2008). Uniquely, we have shown side by side Gdf5 expression across a range of key stages of development in both the forelimb and hindlimb. ...
... Meech et al (2005) reported Gdf5 expression in digit rays at E11.5 and in developing cartilage condensations by E13.5, however we do not see Gdf5 expression exclusively across developing joint sites until E14.5 (Meech et al., 2005;Ota et al., 2007;Seemann et al., 2005). This difference may be due to strain differences in embryonic development which can exhibit differences in gestation periods (Degenkolbe et al., 2013;Huang et al., 2016). Gdf5 is expressed across developing joint sites, including interphalangeal joints, from E14.5 (Chen et al., 2016;Houweling et al., 2001;Huang et al., 2016;Sohaskey et al., 2009). ...
Background
While data regarding expression of limb element and tissue markers during normal mouse limb development exists, few studies show expression patterns in upper and lower limbs throughout key limb development stages. A comparison to normal developmental events is essential when analysing development of the limb in mutant mice models.
Results
Expression patterns of the joint marker Gdf5, tendon and ligament marker Scleraxis, early muscle marker MyoD1, and blood vessel marker Cadherin5 (Cdh5) are presented during the most active phases of embryonic mouse limb patterning. Anti‐neurofilament staining of developing nerves in the fore‐ and hind‐limbs and cartilage formation and progression also is described.
Conclusions
This study demonstrates and describes a range of key morphological markers, and methods, which together, can be used to assess normal and abnormal limb development.
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... GDF5 mutations have previously been associated with several kinds of inherited skeletal diseases. Some of them are autosomal recessive disorders, such as Acromesomelic dysplasia Grebe type, Acromesomelic dysplasia Hunter-Thompson type, and Du Pan syndrome [26,27], whereas others are autosomal dominant disorders, including Proximal symphalangism, Symphalangism proximal 1B, Brachydactyly type A2, Brachydactyly type C, and Multiple synostoses syndrome 2 [5,28]. Brachydactyly type A1 is also associated with GDF5 mutations, which could be autosomal dominant or recessive. ...
Proximal symphalangism (SYM1) is an autosomal dominant disorder, mainly characterized by bony fusions of the proximal phalanges of the hands and feet. GDF5 and NOG were identified to be responsible for SYM1. We have previously reported on a p.Leu373Arg mutation in the GDF5 proregion present in a Chinese family with SYM1. Here, we investigated the effects of the GDF-L373R mutation. The variant caused proteolysis efficiency of GDF5 increased in ATDC5 cells. The variant also caused upregulation of SMAD1/5/8 phosphorylation and increased expression of target genes SMURF1, along with COL2A1 and SOX9 which are factors associated with chondrosis. Furthermore, we developed a human-relevant SYM1 mouse model by making a Gdf5L367R (the orthologous position for L373R in humans) knock-in mouse. Gdf5L367R/+ and Gdf5L367R/L367R mice displayed stiffness and adhesions across the proximal phalanx joint which were in complete accord with SYM1. It was also confirmed the joint formation and development was abnormal in Gdf5L367R/+ and Gdf5L367R/L367R mice, including the failure to develop the primary ossification center and be hypertrophic chondrocytes during embryonic development. This knock-in mouse model offers a tool for assessing the pathogenesis of SYM1 and the function of the GDF5 proregion.
