Limits...
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

Representative images of (A) GFP expression following transfection of BeWo with plasmid alone or (B) CMV-eGFP nanoparticles demonstrate greater transgene expression following complex formation than plasmid alone. (C) GFP expression in BeWo cells after transfection with nanoparticles containing PLAC1-GFP or (D) CyP19a-923-GFP for 4 days, resulted in comparable transgene expression to the global CMV promoter, similar results were seen on 4 different passages. Cell nuclei are stained with Dapi (blue). Proliferation levels (E) and apoptosis levels (F) in BeWo cells were not changed when cells were incubated with nanocomplexes for 48 hours compared to BeWo cells incubated with eGFP plasmid alone for the same time period, mean +/- SEM, n>4 passages per treatment.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4608830&req=5

pone.0140879.g002: Representative images of (A) GFP expression following transfection of BeWo with plasmid alone or (B) CMV-eGFP nanoparticles demonstrate greater transgene expression following complex formation than plasmid alone. (C) GFP expression in BeWo cells after transfection with nanoparticles containing PLAC1-GFP or (D) CyP19a-923-GFP for 4 days, resulted in comparable transgene expression to the global CMV promoter, similar results were seen on 4 different passages. Cell nuclei are stained with Dapi (blue). Proliferation levels (E) and apoptosis levels (F) in BeWo cells were not changed when cells were incubated with nanocomplexes for 48 hours compared to BeWo cells incubated with eGFP plasmid alone for the same time period, mean +/- SEM, n>4 passages per treatment.

Mentions: GFP expression was higher in the BeWo cells incubated with any of the nanoparticles containing trophoblast-specific promoters than those incubated with the uncomplexed DNA. In the BeWo cells, GFP expression levels under the control of the trophoblast-specific promoters, CyP19a-923 and PLAC1, were comparable with the expression seen after transfection with the GFP under the control of the CMV promoter (Fig 2).


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)

Representative images of (A) GFP expression following transfection of BeWo with plasmid alone or (B) CMV-eGFP nanoparticles demonstrate greater transgene expression following complex formation than plasmid alone. (C) GFP expression in BeWo cells after transfection with nanoparticles containing PLAC1-GFP or (D) CyP19a-923-GFP for 4 days, resulted in comparable transgene expression to the global CMV promoter, similar results were seen on 4 different passages. Cell nuclei are stained with Dapi (blue). Proliferation levels (E) and apoptosis levels (F) in BeWo cells were not changed when cells were incubated with nanocomplexes for 48 hours compared to BeWo cells incubated with eGFP plasmid alone for the same time period, mean +/- SEM, n>4 passages per treatment.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140879.g002: Representative images of (A) GFP expression following transfection of BeWo with plasmid alone or (B) CMV-eGFP nanoparticles demonstrate greater transgene expression following complex formation than plasmid alone. (C) GFP expression in BeWo cells after transfection with nanoparticles containing PLAC1-GFP or (D) CyP19a-923-GFP for 4 days, resulted in comparable transgene expression to the global CMV promoter, similar results were seen on 4 different passages. Cell nuclei are stained with Dapi (blue). Proliferation levels (E) and apoptosis levels (F) in BeWo cells were not changed when cells were incubated with nanocomplexes for 48 hours compared to BeWo cells incubated with eGFP plasmid alone for the same time period, mean +/- SEM, n>4 passages per treatment.
Mentions: GFP expression was higher in the BeWo cells incubated with any of the nanoparticles containing trophoblast-specific promoters than those incubated with the uncomplexed DNA. In the BeWo cells, GFP expression levels under the control of the trophoblast-specific promoters, CyP19a-923 and PLAC1, were comparable with the expression seen after transfection with the GFP under the control of the CMV promoter (Fig 2).

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