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Transplantation of Human Neural Progenitor Cells Expressing IGF-1 Enhances Retinal Ganglion Cell Survival.

Ma J, Guo C, Guo C, Sun Y, Liao T, Beattie U, López FJ, Chen DF, Lashkari K - PLoS ONE (2015)

Bottom Line: Application of various IGF-1 signaling blockers or IGF-1 receptor antagonists abrogated these effects.In this model, evaluation of retinal flatmounts and optic nerve cross sections indicated that only hNPIGF-TD cells effectively reduced RGC death and showed a trend to improve optic nerve axonal loss.RT-PCR analysis of retina lysates over time showed that the neurotrophic effects of IGF-1 were also attributed to down-regulation of inflammatory and to some extent, angiogenic pathways.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, 02114, MA, United States of America.

ABSTRACT
We have previously characterized human neuronal progenitor cells (hNP) that can adopt a retinal ganglion cell (RGC)-like morphology within the RGC and nerve fiber layers of the retina. In an effort to determine whether hNPs could be used a candidate cells for targeted delivery of neurotrophic factors (NTFs), we evaluated whether hNPs transfected with an vector that expresses IGF-1 in the form of a fusion protein with tdTomato (TD), would increase RGC survival in vitro and confer neuroprotective effects in a mouse model of glaucoma. RGCs co-cultured with hNPIGF-TD cells displayed enhanced survival, and increased neurite extension and branching as compared to hNPTD or untransfected hNP cells. Application of various IGF-1 signaling blockers or IGF-1 receptor antagonists abrogated these effects. In vivo, using a model of glaucoma we showed that IOP elevation led to reductions in retinal RGC count. In this model, evaluation of retinal flatmounts and optic nerve cross sections indicated that only hNPIGF-TD cells effectively reduced RGC death and showed a trend to improve optic nerve axonal loss. RT-PCR analysis of retina lysates over time showed that the neurotrophic effects of IGF-1 were also attributed to down-regulation of inflammatory and to some extent, angiogenic pathways. This study shows that neuronal progenitor cells that hone into the RGC and nerve fiber layers may be used as vehicles for local production and delivery of a desired NTF. Transplantation of hNPIGF-TD cells improves RGC survival in vitro and protects against RGC loss in a rodent model of glaucoma. Our findings have provided experimental evidence and form the basis for applying cell-based strategies for local delivery of NTFs into the retina. Application of cell-based delivery may be extended to other disease conditions beyond glaucoma.

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Quantification of angiogenic and inflammatory mRNA signals under various experimental conditions (data were presented as mean ± SD).Expression of genes related with angiogenic pathways (A—E), cell apoptosis (F—G) and inflammatory pathways (H—L) are shown. Significant increases in mRNA for typical genes related to angiogenic and inflammatory pathways after microbead injection is shown. These signals are tapered in microbead-injected eyes transplanted with hNPIGF-TD cells. (K) IGF-1 component of the IGF-TD message can be detected in mice transplanted with hNPIGF-TD cells (* P < 0.05 compared to NO group; # P < 0.05 compared to the mice transplanted with hNPIGF-TD cells). Abbreviations: IGF-TD, transplanted hNPIGF-TD cells after microbead injection; TD, transplanted hNPTD cells after microbead injection. hNP, transplanted hNPs after microbead injection; SA, intravitreal saline (no cells) injection after microbead injection; NO, intravitreal saline injection and saline injection into the anterior chamber (no microbead and cell injection). High IOP, elevated intraocular pressure by microbead injection.
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pone.0125695.g007: Quantification of angiogenic and inflammatory mRNA signals under various experimental conditions (data were presented as mean ± SD).Expression of genes related with angiogenic pathways (A—E), cell apoptosis (F—G) and inflammatory pathways (H—L) are shown. Significant increases in mRNA for typical genes related to angiogenic and inflammatory pathways after microbead injection is shown. These signals are tapered in microbead-injected eyes transplanted with hNPIGF-TD cells. (K) IGF-1 component of the IGF-TD message can be detected in mice transplanted with hNPIGF-TD cells (* P < 0.05 compared to NO group; # P < 0.05 compared to the mice transplanted with hNPIGF-TD cells). Abbreviations: IGF-TD, transplanted hNPIGF-TD cells after microbead injection; TD, transplanted hNPTD cells after microbead injection. hNP, transplanted hNPs after microbead injection; SA, intravitreal saline (no cells) injection after microbead injection; NO, intravitreal saline injection and saline injection into the anterior chamber (no microbead and cell injection). High IOP, elevated intraocular pressure by microbead injection.

Mentions: We studied the effects of hNP cell transplantation and secreted proteins, IGF-TD and TD, on gene expression in the host retina. We selected genes that were typically associated with apopototic, inflammatory and angiogenic pathways, listed in Table 1. In the microbead glaucoma groups (groups 2–4; saline, hNP and hNPTD), increased expression of the following genes was detected on day 30, as compared to baseline (all P < 0.05; Fig 7A–7J). These genes included VEGF-A, VEGF-D (increased 13-fold), VEGFR2, CD11b, MFG-E8 (macrophage receptor) and TNF-α.


Transplantation of Human Neural Progenitor Cells Expressing IGF-1 Enhances Retinal Ganglion Cell Survival.

Ma J, Guo C, Guo C, Sun Y, Liao T, Beattie U, López FJ, Chen DF, Lashkari K - PLoS ONE (2015)

Quantification of angiogenic and inflammatory mRNA signals under various experimental conditions (data were presented as mean ± SD).Expression of genes related with angiogenic pathways (A—E), cell apoptosis (F—G) and inflammatory pathways (H—L) are shown. Significant increases in mRNA for typical genes related to angiogenic and inflammatory pathways after microbead injection is shown. These signals are tapered in microbead-injected eyes transplanted with hNPIGF-TD cells. (K) IGF-1 component of the IGF-TD message can be detected in mice transplanted with hNPIGF-TD cells (* P < 0.05 compared to NO group; # P < 0.05 compared to the mice transplanted with hNPIGF-TD cells). Abbreviations: IGF-TD, transplanted hNPIGF-TD cells after microbead injection; TD, transplanted hNPTD cells after microbead injection. hNP, transplanted hNPs after microbead injection; SA, intravitreal saline (no cells) injection after microbead injection; NO, intravitreal saline injection and saline injection into the anterior chamber (no microbead and cell injection). High IOP, elevated intraocular pressure by microbead injection.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414591&req=5

pone.0125695.g007: Quantification of angiogenic and inflammatory mRNA signals under various experimental conditions (data were presented as mean ± SD).Expression of genes related with angiogenic pathways (A—E), cell apoptosis (F—G) and inflammatory pathways (H—L) are shown. Significant increases in mRNA for typical genes related to angiogenic and inflammatory pathways after microbead injection is shown. These signals are tapered in microbead-injected eyes transplanted with hNPIGF-TD cells. (K) IGF-1 component of the IGF-TD message can be detected in mice transplanted with hNPIGF-TD cells (* P < 0.05 compared to NO group; # P < 0.05 compared to the mice transplanted with hNPIGF-TD cells). Abbreviations: IGF-TD, transplanted hNPIGF-TD cells after microbead injection; TD, transplanted hNPTD cells after microbead injection. hNP, transplanted hNPs after microbead injection; SA, intravitreal saline (no cells) injection after microbead injection; NO, intravitreal saline injection and saline injection into the anterior chamber (no microbead and cell injection). High IOP, elevated intraocular pressure by microbead injection.
Mentions: We studied the effects of hNP cell transplantation and secreted proteins, IGF-TD and TD, on gene expression in the host retina. We selected genes that were typically associated with apopototic, inflammatory and angiogenic pathways, listed in Table 1. In the microbead glaucoma groups (groups 2–4; saline, hNP and hNPTD), increased expression of the following genes was detected on day 30, as compared to baseline (all P < 0.05; Fig 7A–7J). These genes included VEGF-A, VEGF-D (increased 13-fold), VEGFR2, CD11b, MFG-E8 (macrophage receptor) and TNF-α.

Bottom Line: Application of various IGF-1 signaling blockers or IGF-1 receptor antagonists abrogated these effects.In this model, evaluation of retinal flatmounts and optic nerve cross sections indicated that only hNPIGF-TD cells effectively reduced RGC death and showed a trend to improve optic nerve axonal loss.RT-PCR analysis of retina lysates over time showed that the neurotrophic effects of IGF-1 were also attributed to down-regulation of inflammatory and to some extent, angiogenic pathways.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, 02114, MA, United States of America.

ABSTRACT
We have previously characterized human neuronal progenitor cells (hNP) that can adopt a retinal ganglion cell (RGC)-like morphology within the RGC and nerve fiber layers of the retina. In an effort to determine whether hNPs could be used a candidate cells for targeted delivery of neurotrophic factors (NTFs), we evaluated whether hNPs transfected with an vector that expresses IGF-1 in the form of a fusion protein with tdTomato (TD), would increase RGC survival in vitro and confer neuroprotective effects in a mouse model of glaucoma. RGCs co-cultured with hNPIGF-TD cells displayed enhanced survival, and increased neurite extension and branching as compared to hNPTD or untransfected hNP cells. Application of various IGF-1 signaling blockers or IGF-1 receptor antagonists abrogated these effects. In vivo, using a model of glaucoma we showed that IOP elevation led to reductions in retinal RGC count. In this model, evaluation of retinal flatmounts and optic nerve cross sections indicated that only hNPIGF-TD cells effectively reduced RGC death and showed a trend to improve optic nerve axonal loss. RT-PCR analysis of retina lysates over time showed that the neurotrophic effects of IGF-1 were also attributed to down-regulation of inflammatory and to some extent, angiogenic pathways. This study shows that neuronal progenitor cells that hone into the RGC and nerve fiber layers may be used as vehicles for local production and delivery of a desired NTF. Transplantation of hNPIGF-TD cells improves RGC survival in vitro and protects against RGC loss in a rodent model of glaucoma. Our findings have provided experimental evidence and form the basis for applying cell-based strategies for local delivery of NTFs into the retina. Application of cell-based delivery may be extended to other disease conditions beyond glaucoma.

Show MeSH
Related in: MedlinePlus