Defining the Earliest Transcriptional Steps of Chondrogenic Progenitor Specification during the Formation of the Digits in the Embryonic Limb

Lorda‐Diez, Carlos I.; Montero, Juan A.; Diaz-Mendoza, Manuel J.; García‐Porrero, Juan A.; Hurlé, Juan M.

doi:10.1371/journal.pone.0024546

Cited by 53 publications

(51 citation statements)

References 86 publications

(103 reference statements)

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“…Interestingly, the pathway involving activin A was also enriched for each comparison analyzed. Less is known about the function of this secreted factor in cartilage biology although there are reports that this TGF-β family member plays a role in developing as well as mature cartilage [102]–[104]. Tables 6 , 7 and 8 shows that Hedgehog and Wnt signaling pathways [105], [106] may be regulated by miRNAs found to be more highly expressed in differentiated or hypertrophic chondrocytes.…”

Section: Discussionmentioning

confidence: 99%

Differentially Expressed MicroRNAs in Chondrocytes from Distinct Regions of Developing Human Cartilage

et al. 2013

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There is compelling in vivo evidence from reports on human genetic mutations and transgenic mice that some microRNAs (miRNAs) play an important functional role in regulating skeletal development and growth. A number of published in vitro studies also point toward a role for miRNAs in controlling chondrocyte gene expression and differentiation. However, information on miRNAs that may regulate a specific phase of chondrocyte differentiation (i.e. production of progenitor, differentiated or hypertrophic chondrocytes) is lacking. To attempt to bridge this knowledge gap, we have investigated miRNA expression patterns in human embryonic cartilage tissue. Specifically, a developmental time point was selected, prior to endochondral ossification in the embryonic limb, to permit analysis of three distinct populations of chondrocytes. The location of chondroprogenitor cells, differentiated chondrocytes and hypertrophic chondrocytes in gestational day 54–56 human embryonic limb tissue sections was confirmed both histologically and by specific collagen expression patterns. Laser capture microdissection was utilized to separate the three chondrocyte populations and a miRNA profiling study was carried out using TaqMan® OpenArray® Human MicroRNA Panels (Applied Biosystems®). Here we report on abundantly expressed miRNAs in human embryonic cartilage tissue and, more importantly, we have identified miRNAs that are significantly differentially expressed between precursor, differentiated and hypertrophic chondrocytes by 2-fold or more. Some of the miRNAs identified in this study have been described in other aspects of cartilage or bone biology, while others have not yet been reported in chondrocytes. Finally, a bioinformatics approach was applied to begin to decipher developmental cellular pathways that may be regulated by groups of differentially expressed miRNAs during distinct stages of chondrogenesis. Data obtained from this work will serve as an important resource of information for the field of cartilage biology and will enhance our understanding of miRNA-driven mechanisms regulating cartilage and endochondral bone development, regeneration and repair.

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Section: Discussionmentioning

confidence: 99%

Differentially Expressed MicroRNAs in Chondrocytes from Distinct Regions of Developing Human Cartilage

et al. 2013

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“…Furthermore, implantation of TGF1-loaded beads into the interdigit mesenchyme of chicken limb buds is able to induce formation of ectopic digit phalanges, i.e. trigger ectopic chondrogenesis (Merino et al, 1999;Lorda-Diez et al, 2011). Genetic analysis of TGF ligands and receptors in mouse embryos has revealed their essential roles during limb long bone, joint and tendon morphogenesis (Sanford et al, 1997;Seo and Serra, 2007)].…”

Section: Introductionmentioning

confidence: 99%

Smad4 is required to induce digit ray primordia and to initiate the aggregation and differentiation of chondrogenic progenitors in mouse limb buds

et al. 2012

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SUMMARYSMAD4 is an essential mediator of canonical TGF/BMP signal transduction and we inactivated Smad4 in mouse limb buds from early stages onward to study its functions in the mesenchyme. While this Smad4 inactivation did not alter the early Sox9 distribution, prefiguring the chondrogenic primordia of the stylopod and zeugopod, it disrupted formation of all Sox9-positive digit ray primordia. Specific inactivation of Smad4 during handplate development pointed to its differential requirement for posterior and anterior digit ray primordia. At the cellular level, Smad4 deficiency blocked the aggregation of Sox9-positive progenitors, thereby preventing chondrogenic differentiation as revealed by absence of collagen type II. The progressive loss of SOX9 due to disrupting digit ray primordia and chondrogenesis was paralleled by alterations in genes marking other lineages. This pointed to a general loss of tissue organization and diversion of mutant cells toward non-specific connective tissue. Conditional inactivation of Bmp2 and Bmp4 indicated that the loss of digit ray primordia and increase in connective tissue were predominantly a consequence of disrupting SMAD4-mediated BMP signal transduction. In summary, our analysis reveals that SMAD4 is required to initiate: (1) formation of the Sox9-positive digit ray primordia; and (2) aggregation and chondrogenic differentiation of all limb skeletal elements.

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“…Since CCN1, CCN3 and mainly CCN2 had already been reported to be important factors regulating chondrocyte differentiation [3,18,19], we evaluated the influence of CCN4 on the chondrogenic process of hBMSCs by inducing CCN4 over-expression using adenovirus encoding CCN4 (adCCN4) as well as by silencing CCN4 gene using siCCN4. AdCCN4 was transduced into hBMSCs [8], and the increase in CCN4 gene expression level was confirmed by real time RT-PCR, in comparison with adCMV-transduced cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Among the TGF-β superfamily, TGF-β1, β2 and β3 are known to play important roles not only in chondrocyte proliferation and differentiation but also in embryonic development of cartilage through activation of the SMAD2/3 signaling pathway; and alterations in TGF-β signaling are known to be involved in the pathogenesis of OA and osteoporosis [1,2]. In early stages of chondrogenesis from mesenchymal stem cell condensation to limb bud formation, TGF-β activates SMAD3, which binds to co-SMAD4, translocates to the nucleus and promotes the expression of Sox9 , the master regulator of chondrogenesis [3]. …”

Section: Introductionmentioning

confidence: 99%

CCN4/WISP-1 positively regulates chondrogenesis by controlling TGF-β3 function

Yoshioka

Ono

Maeda

et al. 2016

Bone

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The CCN family of proteins plays important roles in development and homeostasis of bone and cartilage. To understand the role of CCN4 in chondrogenesis, human bone marrow stromal cells (hBMSCs) were transduced with CCN4 adenovirus (adCCN4) or siRNA to CCN4 (siCCN4) in the presence or absence of transforming growth factor-β3 (TGF-β3). Overexpression of CCN4 enhanced TGF-β3-induced SMAD2/3 phosphorylation and chondrogenesis of hBMSCs in an in vitro assay using a micromass culture model. On the other hand, knockdown of CCN4 inhibited the TGF-β3-induced SMAD2/3 phosphorylation and synthesis of cartilage matrix in micromass cultures of hBMSCs. Immunoprecipitation–western blot analysis revealed that CCN4 bound to TGF-β3 and regulated the ability of TGF-β3 to bind to hBMSCs. In vivo analysis confirmed there was a significant decrease in the gene expression levels of chondrocyte markers in cartilage samples from Ccn4-knock out (KO) mice, compared to those from wild type (WT) control. In order to investigate the regenerative properties of the articular cartilage in Ccn4-KO mice, articular cartilage defects were surgically performed in the knee joints of young mice, and the results showed that the cartilage was partially repaired in WT mice, but not in Ccn4-KO mice. In conclusion, these results show, for the first time, that CCN4 has a positive influence on chondrogenic differentiation by modulating the effects of TGF-β3.

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Defining the Earliest Transcriptional Steps of Chondrogenic Progenitor Specification during the Formation of the Digits in the Embryonic Limb

Cited by 53 publications

References 86 publications

Differentially Expressed MicroRNAs in Chondrocytes from Distinct Regions of Developing Human Cartilage

Differentially Expressed MicroRNAs in Chondrocytes from Distinct Regions of Developing Human Cartilage

Smad4 is required to induce digit ray primordia and to initiate the aggregation and differentiation of chondrogenic progenitors in mouse limb buds

CCN4/WISP-1 positively regulates chondrogenesis by controlling TGF-β3 function

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