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Role of astroglia in Down's syndrome revealed by patient-derived human-induced pluripotent stem cells.

Chen C, Jiang P, Xue H, Peterson SE, Tran HT, McCann AE, Parast MM, Li S, Pleasure DE, Laurent LC, Loring JF, Liu Y, Deng W - Nat Commun (2014)

Bottom Line: DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules.Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo.Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, California 95817, USA [2] Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, California 95817, USA [3] Department of Neurology, Institute of Neurology, Tianjin General Hospital, Tianjin Medical University, Tianjin 300070, China [4].

ABSTRACT
Down's syndrome (DS), caused by trisomy of human chromosome 21, is the most common genetic cause of intellectual disability. Here we use induced pluripotent stem cells (iPSCs) derived from DS patients to identify a role for astrocytes in DS pathogenesis. DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules. Astrocyte-conditioned medium collected from DS astroglia causes toxicity to neurons, and fails to promote neuronal ion channel maturation and synapse formation. Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo. We also observed abnormal gene expression profiles from DS astroglia. Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia. Our studies shed light on the pathogenesis and possible treatment of DS by targeting astrocytes with a clinically available drug.

No MeSH data available.


Related in: MedlinePlus

The effects of DS astroglia on the DS NPC differentiation and DS neuronsurvival.(a) Representatives of DS NPC differentiated into βIII-tubulin+ neurons andS100B+ astrogliaunder spontaneous differentiation condition in the presence of DS ACM,control (Cont) ACM, DS S100BsiRNA ACM and DS-Mino ACM. (b) βIII-tubulin andactivated caspase3co-staining of DS neurons cultured with different ACM. Blue, 4′,6-diamidino-2-phenylindoledihydrochloride(DAPI)-stained nuclei. Scale bars,50 μm. (c,d) Quantification of pooleddata showing the percentage of βIII-tubulin+ neurons and S100B+ astroglia derived from DSNPCs (n=3–4 from each cell line), and the length of thelongest neurites of neurons (n=10 from each cell line) underspontaneous differentiation conditions in the presence of different ACM.(e) Quantification of pooled data showing the percentage ofβIII-tubulin+ and activated caspase3+ cells among the groupswith different treatments (n=3–5 from each cell line).One-way analysis of variance test, *P<0.05 and**P<0.01. Data are presented as mean±s.e.m. NS, notsignificant.
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f3: The effects of DS astroglia on the DS NPC differentiation and DS neuronsurvival.(a) Representatives of DS NPC differentiated into βIII-tubulin+ neurons andS100B+ astrogliaunder spontaneous differentiation condition in the presence of DS ACM,control (Cont) ACM, DS S100BsiRNA ACM and DS-Mino ACM. (b) βIII-tubulin andactivated caspase3co-staining of DS neurons cultured with different ACM. Blue, 4′,6-diamidino-2-phenylindoledihydrochloride(DAPI)-stained nuclei. Scale bars,50 μm. (c,d) Quantification of pooleddata showing the percentage of βIII-tubulin+ neurons and S100B+ astroglia derived from DSNPCs (n=3–4 from each cell line), and the length of thelongest neurites of neurons (n=10 from each cell line) underspontaneous differentiation conditions in the presence of different ACM.(e) Quantification of pooled data showing the percentage ofβIII-tubulin+ and activated caspase3+ cells among the groupswith different treatments (n=3–5 from each cell line).One-way analysis of variance test, *P<0.05 and**P<0.01. Data are presented as mean±s.e.m. NS, notsignificant.

Mentions: Accumulative studies have shown that astrocytes promote neuronal differentiationfrom NPCs, and regulate neurite outgrowth and synaptogenesis via releasingsoluble factors1732333435. To further test the hypothesisthat DS astroglia contributes to the reduced neurogenesis of NPCs, we fed NPCsderived from DS iPSCs with concentrated DS ACM or control ACM for 1 week. Thepercentage of βIII-tubulin+ neurons and S100B+ astroglia generated from DS NPCswas then quantified. As shown in Fig. 3a,c, similar to DSNPCs cultured under spontaneous differentiation condition (Fig.1h,i), DS NPCs cultured with DS ACM gave rise to low percentage ofβIII-tubulin+neurons (19.7±1.3%) and high percentage of S100B+ astroglia (78.2±1.2%;n=3–4 for each cell line). Interestingly, the addition ofcontrol ACM restored the reduced neurogenesis of DS NPCs (33.0±2.4% and66.3±2.5% for βIII-tubulin+neurons and S100B+ astroglia, respectively;P<0.01; n=3–4 for each cell line). Moreover,we analysed the neurite length of the differentiated DS neurons and found thatthe average length of the longest neurites of the neurons generated from NPCs inDS ACM (Fig. 3d;65.8±4.7 μm) was significantly shorter than thoseof the neurons generated from NPCs in control ACM (Fig.3d, 81.0±2.7 μm; P<0.05;n=10 from each cell line). To further examine whether theoverexpression of S100B in DSastroglia contributed to these effects, we fed the DS NPCs with ACM collectedfrom DS astroglia transfected with S100B siRNA (DS S100BsiRNA ACM). Compared with theNPCs cultured with DS ACM, the addition of DS S100BsiRNA ACM rescued the reducedneurogenesis from DS NPCs (Fig. 3a,c, 28.8±1.3%and 70.3±2.6% for βIII-tubulin+ neurons and S100B+ astroglia, respectively;P<0.05; n=3-4 from each cell line). However, theneurite length was not significantly different between the cells fed with DS ACMand DS S100BsiRNA ACM (Fig. 3d,71.2±2.1 μm; P>0.05; n=10from each cell line). Consistent with the effect of minocycline on correcting the geneexpression of DS astroglia, the addition of ACM collected from minocycline-treated DS astroglia(DS-Mino ACM) had effectssimilar to the addition of control ACM (Fig. 3c,29.4±2.5% and 70.1±2.2% for βIII-tubulin+ neurons andS100B+ astroglia,respectively; n=3–4 from each cell line, and Fig. 3d, 81.9±2.3 μm for the averagelength of the longest neurites; n=10 from each cell line).


Role of astroglia in Down's syndrome revealed by patient-derived human-induced pluripotent stem cells.

Chen C, Jiang P, Xue H, Peterson SE, Tran HT, McCann AE, Parast MM, Li S, Pleasure DE, Laurent LC, Loring JF, Liu Y, Deng W - Nat Commun (2014)

The effects of DS astroglia on the DS NPC differentiation and DS neuronsurvival.(a) Representatives of DS NPC differentiated into βIII-tubulin+ neurons andS100B+ astrogliaunder spontaneous differentiation condition in the presence of DS ACM,control (Cont) ACM, DS S100BsiRNA ACM and DS-Mino ACM. (b) βIII-tubulin andactivated caspase3co-staining of DS neurons cultured with different ACM. Blue, 4′,6-diamidino-2-phenylindoledihydrochloride(DAPI)-stained nuclei. Scale bars,50 μm. (c,d) Quantification of pooleddata showing the percentage of βIII-tubulin+ neurons and S100B+ astroglia derived from DSNPCs (n=3–4 from each cell line), and the length of thelongest neurites of neurons (n=10 from each cell line) underspontaneous differentiation conditions in the presence of different ACM.(e) Quantification of pooled data showing the percentage ofβIII-tubulin+ and activated caspase3+ cells among the groupswith different treatments (n=3–5 from each cell line).One-way analysis of variance test, *P<0.05 and**P<0.01. Data are presented as mean±s.e.m. NS, notsignificant.
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Related In: Results  -  Collection

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f3: The effects of DS astroglia on the DS NPC differentiation and DS neuronsurvival.(a) Representatives of DS NPC differentiated into βIII-tubulin+ neurons andS100B+ astrogliaunder spontaneous differentiation condition in the presence of DS ACM,control (Cont) ACM, DS S100BsiRNA ACM and DS-Mino ACM. (b) βIII-tubulin andactivated caspase3co-staining of DS neurons cultured with different ACM. Blue, 4′,6-diamidino-2-phenylindoledihydrochloride(DAPI)-stained nuclei. Scale bars,50 μm. (c,d) Quantification of pooleddata showing the percentage of βIII-tubulin+ neurons and S100B+ astroglia derived from DSNPCs (n=3–4 from each cell line), and the length of thelongest neurites of neurons (n=10 from each cell line) underspontaneous differentiation conditions in the presence of different ACM.(e) Quantification of pooled data showing the percentage ofβIII-tubulin+ and activated caspase3+ cells among the groupswith different treatments (n=3–5 from each cell line).One-way analysis of variance test, *P<0.05 and**P<0.01. Data are presented as mean±s.e.m. NS, notsignificant.
Mentions: Accumulative studies have shown that astrocytes promote neuronal differentiationfrom NPCs, and regulate neurite outgrowth and synaptogenesis via releasingsoluble factors1732333435. To further test the hypothesisthat DS astroglia contributes to the reduced neurogenesis of NPCs, we fed NPCsderived from DS iPSCs with concentrated DS ACM or control ACM for 1 week. Thepercentage of βIII-tubulin+ neurons and S100B+ astroglia generated from DS NPCswas then quantified. As shown in Fig. 3a,c, similar to DSNPCs cultured under spontaneous differentiation condition (Fig.1h,i), DS NPCs cultured with DS ACM gave rise to low percentage ofβIII-tubulin+neurons (19.7±1.3%) and high percentage of S100B+ astroglia (78.2±1.2%;n=3–4 for each cell line). Interestingly, the addition ofcontrol ACM restored the reduced neurogenesis of DS NPCs (33.0±2.4% and66.3±2.5% for βIII-tubulin+neurons and S100B+ astroglia, respectively;P<0.01; n=3–4 for each cell line). Moreover,we analysed the neurite length of the differentiated DS neurons and found thatthe average length of the longest neurites of the neurons generated from NPCs inDS ACM (Fig. 3d;65.8±4.7 μm) was significantly shorter than thoseof the neurons generated from NPCs in control ACM (Fig.3d, 81.0±2.7 μm; P<0.05;n=10 from each cell line). To further examine whether theoverexpression of S100B in DSastroglia contributed to these effects, we fed the DS NPCs with ACM collectedfrom DS astroglia transfected with S100B siRNA (DS S100BsiRNA ACM). Compared with theNPCs cultured with DS ACM, the addition of DS S100BsiRNA ACM rescued the reducedneurogenesis from DS NPCs (Fig. 3a,c, 28.8±1.3%and 70.3±2.6% for βIII-tubulin+ neurons and S100B+ astroglia, respectively;P<0.05; n=3-4 from each cell line). However, theneurite length was not significantly different between the cells fed with DS ACMand DS S100BsiRNA ACM (Fig. 3d,71.2±2.1 μm; P>0.05; n=10from each cell line). Consistent with the effect of minocycline on correcting the geneexpression of DS astroglia, the addition of ACM collected from minocycline-treated DS astroglia(DS-Mino ACM) had effectssimilar to the addition of control ACM (Fig. 3c,29.4±2.5% and 70.1±2.2% for βIII-tubulin+ neurons andS100B+ astroglia,respectively; n=3–4 from each cell line, and Fig. 3d, 81.9±2.3 μm for the averagelength of the longest neurites; n=10 from each cell line).

Bottom Line: DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules.Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo.Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, California 95817, USA [2] Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, California 95817, USA [3] Department of Neurology, Institute of Neurology, Tianjin General Hospital, Tianjin Medical University, Tianjin 300070, China [4].

ABSTRACT
Down's syndrome (DS), caused by trisomy of human chromosome 21, is the most common genetic cause of intellectual disability. Here we use induced pluripotent stem cells (iPSCs) derived from DS patients to identify a role for astrocytes in DS pathogenesis. DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules. Astrocyte-conditioned medium collected from DS astroglia causes toxicity to neurons, and fails to promote neuronal ion channel maturation and synapse formation. Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo. We also observed abnormal gene expression profiles from DS astroglia. Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia. Our studies shed light on the pathogenesis and possible treatment of DS by targeting astrocytes with a clinically available drug.

No MeSH data available.


Related in: MedlinePlus