Cotransfected human chondrocytes: over-expression of IGF-I and SOX9 enhances the synthesis of cartilage matrix components collagen-II and glycosaminoglycans

Simental‐Mendía, Mario; Lara‐Arias, Jorge; Álvarez-Lozano, Eduardo; Said‐Fernández, Salvador; Soto‐Domínguez, Adolfo; Padilla‐Rivas, Gerardo R.; Martı́nez-Rodrı́guez, Herminia Guadalupe

doi:10.1590/1414-431x20154732

Cited by 11 publications

(15 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Meanwhile, we verified that upregulation of miR-195 suppressed FGF-18 expression and the activity level of its downstream FGFR3/ FGFR3 p-Tyr724 , ERK1/2/ERK1/2 p-Thr202/Tyr204 , and SOX9. SOX9, which was well accepted as the upstream regulator of Col2a1 and aggrecan [62,63,64,65,66], was reported as being regulated by FGF-18 according to former studies [67,68]. Furthermore, in the present study, we also showed that ectopic miR-195 could correspondingly regulate the expression of FGF-18 and its downstream SOX9 and further effect the expression of Col2a1 and aggrecan.…”

Section: Discussionsupporting

confidence: 78%

Decrease of miR-195 Promotes Chondrocytes Proliferation and Maintenance of Chondrogenic Phenotype via Targeting FGF-18 Pathway

Wang

Yang

Liu

et al. 2017

IJMS

View full text Add to dashboard Cite

Slow growth and rapid loss of chondrogenic phenotypes are the major problems affecting chronic cartilage lesions. The role of microRNA-195 (miR-195) and its detailed working mechanism in the fore-mentioned process remains unknown. Fibroblastic growth factor 18 (FGF-18) plays a key role in cartilage homeostasis; whether miR-195 could regulate FGF-18 and its downstream signal pathway in chondrocyte proliferation and maintenance of chondrogenic phenotypes still remains unclear. The present research shows elevated miR-195 but depressed FGF-18 expressed in joint fluid specimens of 20 patients with chronic cartilage lesions and in CH1M and CH3M chondrocytes when compared with that in joint fluid specimens without cartilage lesions and in CH1W and CH2W chondrocytes, respectively. The following loss of function test revealed that downregulation of miR-195 by transfection of miR-195 inhibitors promoted chondrocyte proliferation and expression of a type II collagen α I chain (Col2a1)/aggrecan. Through the online informatics analysis we theoretically predicted that miR-195 could bind to a FGF-18 3′ untranslated region (3′UTR), also, we verified that a miR-195 could regulate the FGF-18 and its downstream pathway. The constructed dual luciferase assay further confirmed that FGF-18 was a direct target of miR-195. The executed anti-sense experiment displayed that miR-195 could regulate chondrocyte proliferation and Col2a1/aggrecan expression via the FGF-18 pathway. Finally, through an in vivo anterior cruciate ligament transection (ACLT) model, downregulation of miR-195 presented a significantly protective effect on chronic cartilage lesions. Evaluating all of the outcomes of the current research revealed that a decrease of miR-195 protected chronic cartilage lesions by promoting chondrocyte proliferation and maintenance of chondrogenic phenotypes via the targeting of the FGF-18 pathway and that the miR-195/FGF-18 axis could be a potential target in the treatment of cartilage lesions.

show abstract

Section: Discussionsupporting

confidence: 78%

Decrease of miR-195 Promotes Chondrocytes Proliferation and Maintenance of Chondrogenic Phenotype via Targeting FGF-18 Pathway

Wang

Yang

Liu

et al. 2017

IJMS

View full text Add to dashboard Cite

show abstract

“…Naturally, mutations affecting the function of these transcription factors would affect the integrity of the ECM by altering its composition. Most of the transcription factors included on the list fall in this category, often regulating directly the expression of collagen, fibronectin, or cadherin genes (Bi et al, ; Hsieh et al, ; Cho et al, ; Papangeli and Scambler, ; Simental‐Mendia et al, ; Horie et al, ; Yoh et al, ). Transcriptional control might be contextual as seen with the Saethre–Chotzen gene TWIST1 (101400) that can both upregulate and downregulate collagen expression (Fang et al, ; Garcia‐Palmero et al, ).…”

Section: Ecm Expression During Palatal Growthmentioning

confidence: 99%

Extracellular Matrix in Secondary Palate Development

Logan

Ruest

Benson

et al. 2019

The Anatomical Record

View full text Add to dashboard Cite

The secondary palate arises from outgrowths of epithelia-covered embryonic mesenchyme that grow from the maxillary prominence, remodel to meet over the tongue, and fuse at the midline. These events require the coordination of cell proliferation, migration, and gene expression, all of which take place in the context of the extracellular matrix (ECM). Palatal cells generate their ECM, and then stiffen, degrade, or otherwise modify its properties to achieve the required cell movement and organization during palatogenesis. The ECM, in turn, acts on the cells through their matrix receptors to change their gene expression and thus their phenotype. The number of ECM-related gene mutations that cause cleft palate in mice and humans is a testament to the crucial role the matrix plays in palate development and a reminder that understanding that role is vital to our progress in treating palate deformities. This article will review the known ECM constituents at each stage of palatogenesis, the mechanisms of tissue reorganization and cell migration through the palatal ECM, the reciprocal relationship between the ECM and gene expression, and human syndromes with cleft palate that arise from mutations of ECM proteins and their regulators. Anat Rec, 303:1543Rec, 303: -1556Rec, 303: , 2020

show abstract

“…In their study, the resulting heterogeneity in the expression of transfected genes was analyzed on the single-cell level for the fast and efficient characterization and optimization of synthetic genetic systems and circuits [ 38 – 40 ]. Functional relevance of the integrated co-transfection method has been addressed in a study by Mendia et al, suggesting the highest cartilage matrix deposition secreted by integrated co-transfected human chondrocytes expressing both IGF-I and SOX9 [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Strategies for simultaneous and successive delivery of RNA

2020

View full text Add to dashboard Cite

Advanced non-viral gene delivery experiments often require co-delivery of multiple nucleic acids. Therefore, the availability of reliable and robust co-transfection methods and defined selection criteria for their use in, e.g., expression of multimeric proteins or mixed RNA/DNA delivery is of utmost importance. Here, we investigated different co- and successive transfection approaches, with particular focus on in vitro transcribed messenger RNA (IVT-mRNA). Expression levels and patterns of two fluorescent protein reporters were determined, using different IVT-mRNA doses, carriers, and cell types. Quantitative parameters determining the efficiency of co-delivery were analyzed for IVT-mRNAs premixed before nanocarrier formation (integrated co-transfection) and when simultaneously transfecting cells with separately formed nanocarriers (parallel co-transfection), which resulted in a much higher level of expression heterogeneity for the two reporters. Successive delivery of mRNA revealed a lower transfection efficiency in the second transfection round. All these differences proved to be more pronounced for low mRNA doses. Concurrent delivery of siRNA with mRNA also indicated the highest co-transfection efficiency for integrated method. However, the maximum efficacy was shown for successive delivery, due to the kinetically different peak output for the two discretely operating entities. Our findings provide guidance for selection of the co-delivery method best suited to accommodate experimental requirements, highlighting in particular the nucleic acid dose-response dependence on co-delivery on the single-cell level.

show abstract

Cotransfected human chondrocytes: over-expression of IGF-I and SOX9 enhances the synthesis of cartilage matrix components collagen-II and glycosaminoglycans

Cited by 11 publications

References 26 publications

Decrease of miR-195 Promotes Chondrocytes Proliferation and Maintenance of Chondrogenic Phenotype via Targeting FGF-18 Pathway

Decrease of miR-195 Promotes Chondrocytes Proliferation and Maintenance of Chondrogenic Phenotype via Targeting FGF-18 Pathway

Extracellular Matrix in Secondary Palate Development

Strategies for simultaneous and successive delivery of RNA

Contact Info

Product

Resources

About