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Development of Non-Viral, Trophoblast-Specific Gene Delivery for Placental Therapy.

Abd Ellah N, Taylor L, Troja W, Owens K, Ayres N, Pauletti G, Jones H - PLoS ONE (2015)

Bottom Line: Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease.To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta.IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth.

View Article: PubMed Central - PubMed

Affiliation: James L. Winkle College of Pharmacy, University of Cincinnati, Cincinnati, OH, 45267, United States of America; Faculty of Pharmacy, Assiut University, 71515, Assiut, Arab Republic of Egypt.

ABSTRACT
Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease. Placental insufficiency leading to intrauterine growth restriction (IUGR) contributes to the prevalence of diseases with developmental origins. Currently there are no therapies for IUGR or placental insufficiency. To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta. IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth. Delivery of genes to a model of human trophoblast and mouse placenta was achieved using a diblock copolymer (pHPMA-b-pDMAEMA) complexed to hIGF-1 plasmid DNA under the control of trophoblast-specific promoters (Cyp19a or PLAC1). Transfection efficiency of pEGFP-C1-containing nanocarriers in BeWo cells and non-trophoblast cells was visually assessed via fluorescence microscopy. In vivo transfection and functionality was assessed by direct placental-injection into a mouse model of IUGR. Complexes formed using pHPMA-b-pDMAEMA and CYP19a-923 or PLAC1-modified plasmids induce trophoblast-selective transgene expression in vitro, and placental injection of PLAC1-hIGF-1 produces measurable RNA expression and alleviates IUGR in our mouse model, consequently representing innovative building blocks towards human placental gene therapies.

No MeSH data available.


Related in: MedlinePlus

GFP expression in Human uterine fibroblasts after transfection with nanoparticles containing (A) CMV-GFP, (B) PLAC1-GFP or (C) CyP19a-923-GFP. HEK293 cells transfected with (D) CMV-GFP, (E) PLAC1-GFP or (F) CyP19a-923-GFP demonstrated low but visible transgene expression under the CMV promoter but no GFP expression with the trophoblast-specific promoters. GFP expression in HPMVECs cells after transfection with nanoparticles containing (G) CMV-GFP, (H) PLAC1-GFP or (I) CyP19a-923-GFP. Representative images seen on 4 different passages for each cell type.Cell nuclei are stained with Dapi (blue).
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pone.0140879.g003: GFP expression in Human uterine fibroblasts after transfection with nanoparticles containing (A) CMV-GFP, (B) PLAC1-GFP or (C) CyP19a-923-GFP. HEK293 cells transfected with (D) CMV-GFP, (E) PLAC1-GFP or (F) CyP19a-923-GFP demonstrated low but visible transgene expression under the CMV promoter but no GFP expression with the trophoblast-specific promoters. GFP expression in HPMVECs cells after transfection with nanoparticles containing (G) CMV-GFP, (H) PLAC1-GFP or (I) CyP19a-923-GFP. Representative images seen on 4 different passages for each cell type.Cell nuclei are stained with Dapi (blue).

Mentions: Expression of GFP was seen in human uterine fibroblasts following incubation with nanoparticles containing GFP under the control of the CMV promoter (Fig 3A), but no GFP expression was observed in fibroblasts when the trophoblast-specific promoters were used (Figs 3B & 2C). Similarly, Human Placental Microvascular Endothelial cells expressed GFP following transfection with the nanoparticle containing the CMV-GFP plasmid as seen in Fig 3G but no GFP was expressed following transfection with either PLAC1-GFP or the CyP-19-923-GFP (Figs 3H & 2I). Interestingly, HEK293 cells showed minimal GFP expression following transfection with the CMV-GFP nanoparticles (Fig 3D) but again showed no expression when the trophoblast-specific promoters were used (Figs 3E & 2F).


Development of Non-Viral, Trophoblast-Specific Gene Delivery for Placental Therapy.

Abd Ellah N, Taylor L, Troja W, Owens K, Ayres N, Pauletti G, Jones H - PLoS ONE (2015)

GFP expression in Human uterine fibroblasts after transfection with nanoparticles containing (A) CMV-GFP, (B) PLAC1-GFP or (C) CyP19a-923-GFP. HEK293 cells transfected with (D) CMV-GFP, (E) PLAC1-GFP or (F) CyP19a-923-GFP demonstrated low but visible transgene expression under the CMV promoter but no GFP expression with the trophoblast-specific promoters. GFP expression in HPMVECs cells after transfection with nanoparticles containing (G) CMV-GFP, (H) PLAC1-GFP or (I) CyP19a-923-GFP. Representative images seen on 4 different passages for each cell type.Cell nuclei are stained with Dapi (blue).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4608830&req=5

pone.0140879.g003: GFP expression in Human uterine fibroblasts after transfection with nanoparticles containing (A) CMV-GFP, (B) PLAC1-GFP or (C) CyP19a-923-GFP. HEK293 cells transfected with (D) CMV-GFP, (E) PLAC1-GFP or (F) CyP19a-923-GFP demonstrated low but visible transgene expression under the CMV promoter but no GFP expression with the trophoblast-specific promoters. GFP expression in HPMVECs cells after transfection with nanoparticles containing (G) CMV-GFP, (H) PLAC1-GFP or (I) CyP19a-923-GFP. Representative images seen on 4 different passages for each cell type.Cell nuclei are stained with Dapi (blue).
Mentions: Expression of GFP was seen in human uterine fibroblasts following incubation with nanoparticles containing GFP under the control of the CMV promoter (Fig 3A), but no GFP expression was observed in fibroblasts when the trophoblast-specific promoters were used (Figs 3B & 2C). Similarly, Human Placental Microvascular Endothelial cells expressed GFP following transfection with the nanoparticle containing the CMV-GFP plasmid as seen in Fig 3G but no GFP was expressed following transfection with either PLAC1-GFP or the CyP-19-923-GFP (Figs 3H & 2I). Interestingly, HEK293 cells showed minimal GFP expression following transfection with the CMV-GFP nanoparticles (Fig 3D) but again showed no expression when the trophoblast-specific promoters were used (Figs 3E & 2F).

Bottom Line: Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease.To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta.IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth.

View Article: PubMed Central - PubMed

Affiliation: James L. Winkle College of Pharmacy, University of Cincinnati, Cincinnati, OH, 45267, United States of America; Faculty of Pharmacy, Assiut University, 71515, Assiut, Arab Republic of Egypt.

ABSTRACT
Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease. Placental insufficiency leading to intrauterine growth restriction (IUGR) contributes to the prevalence of diseases with developmental origins. Currently there are no therapies for IUGR or placental insufficiency. To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta. IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth. Delivery of genes to a model of human trophoblast and mouse placenta was achieved using a diblock copolymer (pHPMA-b-pDMAEMA) complexed to hIGF-1 plasmid DNA under the control of trophoblast-specific promoters (Cyp19a or PLAC1). Transfection efficiency of pEGFP-C1-containing nanocarriers in BeWo cells and non-trophoblast cells was visually assessed via fluorescence microscopy. In vivo transfection and functionality was assessed by direct placental-injection into a mouse model of IUGR. Complexes formed using pHPMA-b-pDMAEMA and CYP19a-923 or PLAC1-modified plasmids induce trophoblast-selective transgene expression in vitro, and placental injection of PLAC1-hIGF-1 produces measurable RNA expression and alleviates IUGR in our mouse model, consequently representing innovative building blocks towards human placental gene therapies.

No MeSH data available.


Related in: MedlinePlus