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MiR-30a-3p negatively regulates BAFF synthesis in systemic sclerosis and rheumatoid arthritis fibroblasts.

Alsaleh G, François A, Philippe L, Gong YZ, Bahram S, Cetin S, Pfeffer S, Gottenberg JE, Wachsmann D, Georgel P, Sibilia J - PLoS ONE (2014)

Bottom Line: Moreover, we demonstrated that transfecting miR-30a-3p mimic in Poly(I:C)- and IFN-γ-activated RAFLS and SScHDF showed a strong decrease on BAFF synthesis and release and thus B cells survival in our model.Transfection of miR-30a-3p antisense in Poly(I:C)- and IFN-γ-activated NFLS and NHDF upregulated BAFF secretion, confirming that this microRNA is a basal repressors of BAFF expression in cells from healthy donors.Our data suggest a critical role of miR-30a-3p in the regulation of BAFF expression, which could have a major impact in the regulation of the autoimmune responses occurring in RA and SSc.

View Article: PubMed Central - PubMed

Affiliation: Immunorhumatologie moléculaire, INSERM UMR S_1109, Centre de Recherche en Immunologie et Hématologie, Faculté de Médecine, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg, France and Service de Rhumatologie, Centre National de Référence pour les Maladies Systémiques Autoimmunes Rares, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.

ABSTRACT
We evaluated micro (mi) RNA-mediated regulation of BAFF expression in fibroblasts using two concomitant models: (i) synovial fibroblasts (FLS) isolated from healthy controls (N) or Rheumatoid Arthritis (RA) patients; (ii) human dermal fibroblasts (HDF) isolated from healthy controls (N) or Systemic Sclerosis (SSc) patients. Using RT-qPCR and ELISA, we first showed that SScHDF synthesized and released BAFF in response to Poly(I:C) or IFN-γ treatment, as previously observed in RAFLS, whereas NHDF released BAFF preferentially in response to IFN-γ. Next, we demonstrated that miR-30a-3p expression was down regulated in RAFLS and SScHDF stimulated with Poly(I:C) or IFN-γ. Moreover, we demonstrated that transfecting miR-30a-3p mimic in Poly(I:C)- and IFN-γ-activated RAFLS and SScHDF showed a strong decrease on BAFF synthesis and release and thus B cells survival in our model. Interestingly, FLS and HDF isolated from healthy subjects express higher levels of miR-30a-3p and lower levels of BAFF than RAFLS and SScHDF. Transfection of miR-30a-3p antisense in Poly(I:C)- and IFN-γ-activated NFLS and NHDF upregulated BAFF secretion, confirming that this microRNA is a basal repressors of BAFF expression in cells from healthy donors. Our data suggest a critical role of miR-30a-3p in the regulation of BAFF expression, which could have a major impact in the regulation of the autoimmune responses occurring in RA and SSc.

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miR-30a-3p expression is down-regulated in Poly (I:C)- and IFN-γ-stimulated RAFLS and SScHDF.A. miR-30a-3p is predicted to target BAFF 3′ UTR mRNA. B, C. miR-30a-3p expression was determined by RT-qPCR in RAFLS (n = 4) (B) and SScHDF (n = 4) (C) stimulated with Poly (I:C) (10 µg/mL) or IFN-γ (0.1 or 5 ng/mL) for 6 h, 48 h and 72 h. Results were normalized to U6snRNA and expressed as fold change compared with samples from cells incubated in medium. Data are expressed as the mean of triplicate samples ± SEM. *p<0.05, ***p<0.001.
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pone-0111266-g002: miR-30a-3p expression is down-regulated in Poly (I:C)- and IFN-γ-stimulated RAFLS and SScHDF.A. miR-30a-3p is predicted to target BAFF 3′ UTR mRNA. B, C. miR-30a-3p expression was determined by RT-qPCR in RAFLS (n = 4) (B) and SScHDF (n = 4) (C) stimulated with Poly (I:C) (10 µg/mL) or IFN-γ (0.1 or 5 ng/mL) for 6 h, 48 h and 72 h. Results were normalized to U6snRNA and expressed as fold change compared with samples from cells incubated in medium. Data are expressed as the mean of triplicate samples ± SEM. *p<0.05, ***p<0.001.

Mentions: Interestingly, we noted that both miR-30a-3p and miR-30e-3p (data not shown), were significantly down-regulated in Poly (I:C)- and IFN-γ-activated, RAFLS (Figure 2B) and SScHDF (figure 2C) 48 h and 72 h after stimulation. These data, together with those illustrated in figure 1, indicate that BAFF transcripts and miR-30a-3p exhibit opposite expression patterns, therefore suggesting potential interactions. Given the strong similarities between miR-30a-3p and miR-30e-3p, we decided to focus on miR-30a-3p.


MiR-30a-3p negatively regulates BAFF synthesis in systemic sclerosis and rheumatoid arthritis fibroblasts.

Alsaleh G, François A, Philippe L, Gong YZ, Bahram S, Cetin S, Pfeffer S, Gottenberg JE, Wachsmann D, Georgel P, Sibilia J - PLoS ONE (2014)

miR-30a-3p expression is down-regulated in Poly (I:C)- and IFN-γ-stimulated RAFLS and SScHDF.A. miR-30a-3p is predicted to target BAFF 3′ UTR mRNA. B, C. miR-30a-3p expression was determined by RT-qPCR in RAFLS (n = 4) (B) and SScHDF (n = 4) (C) stimulated with Poly (I:C) (10 µg/mL) or IFN-γ (0.1 or 5 ng/mL) for 6 h, 48 h and 72 h. Results were normalized to U6snRNA and expressed as fold change compared with samples from cells incubated in medium. Data are expressed as the mean of triplicate samples ± SEM. *p<0.05, ***p<0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4216016&req=5

pone-0111266-g002: miR-30a-3p expression is down-regulated in Poly (I:C)- and IFN-γ-stimulated RAFLS and SScHDF.A. miR-30a-3p is predicted to target BAFF 3′ UTR mRNA. B, C. miR-30a-3p expression was determined by RT-qPCR in RAFLS (n = 4) (B) and SScHDF (n = 4) (C) stimulated with Poly (I:C) (10 µg/mL) or IFN-γ (0.1 or 5 ng/mL) for 6 h, 48 h and 72 h. Results were normalized to U6snRNA and expressed as fold change compared with samples from cells incubated in medium. Data are expressed as the mean of triplicate samples ± SEM. *p<0.05, ***p<0.001.
Mentions: Interestingly, we noted that both miR-30a-3p and miR-30e-3p (data not shown), were significantly down-regulated in Poly (I:C)- and IFN-γ-activated, RAFLS (Figure 2B) and SScHDF (figure 2C) 48 h and 72 h after stimulation. These data, together with those illustrated in figure 1, indicate that BAFF transcripts and miR-30a-3p exhibit opposite expression patterns, therefore suggesting potential interactions. Given the strong similarities between miR-30a-3p and miR-30e-3p, we decided to focus on miR-30a-3p.

Bottom Line: Moreover, we demonstrated that transfecting miR-30a-3p mimic in Poly(I:C)- and IFN-γ-activated RAFLS and SScHDF showed a strong decrease on BAFF synthesis and release and thus B cells survival in our model.Transfection of miR-30a-3p antisense in Poly(I:C)- and IFN-γ-activated NFLS and NHDF upregulated BAFF secretion, confirming that this microRNA is a basal repressors of BAFF expression in cells from healthy donors.Our data suggest a critical role of miR-30a-3p in the regulation of BAFF expression, which could have a major impact in the regulation of the autoimmune responses occurring in RA and SSc.

View Article: PubMed Central - PubMed

Affiliation: Immunorhumatologie moléculaire, INSERM UMR S_1109, Centre de Recherche en Immunologie et Hématologie, Faculté de Médecine, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg, France and Service de Rhumatologie, Centre National de Référence pour les Maladies Systémiques Autoimmunes Rares, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.

ABSTRACT
We evaluated micro (mi) RNA-mediated regulation of BAFF expression in fibroblasts using two concomitant models: (i) synovial fibroblasts (FLS) isolated from healthy controls (N) or Rheumatoid Arthritis (RA) patients; (ii) human dermal fibroblasts (HDF) isolated from healthy controls (N) or Systemic Sclerosis (SSc) patients. Using RT-qPCR and ELISA, we first showed that SScHDF synthesized and released BAFF in response to Poly(I:C) or IFN-γ treatment, as previously observed in RAFLS, whereas NHDF released BAFF preferentially in response to IFN-γ. Next, we demonstrated that miR-30a-3p expression was down regulated in RAFLS and SScHDF stimulated with Poly(I:C) or IFN-γ. Moreover, we demonstrated that transfecting miR-30a-3p mimic in Poly(I:C)- and IFN-γ-activated RAFLS and SScHDF showed a strong decrease on BAFF synthesis and release and thus B cells survival in our model. Interestingly, FLS and HDF isolated from healthy subjects express higher levels of miR-30a-3p and lower levels of BAFF than RAFLS and SScHDF. Transfection of miR-30a-3p antisense in Poly(I:C)- and IFN-γ-activated NFLS and NHDF upregulated BAFF secretion, confirming that this microRNA is a basal repressors of BAFF expression in cells from healthy donors. Our data suggest a critical role of miR-30a-3p in the regulation of BAFF expression, which could have a major impact in the regulation of the autoimmune responses occurring in RA and SSc.

Show MeSH
Related in: MedlinePlus