Peri- and postnatal effects of prenatal adenoviral VEGF gene therapy in growth-restricted sheep

David J Carr, Jacqueline M Wallace, Raymond P Aitken, John S Milne, John F. Martin, Ian C. Zachary, Donald M. Peebles, Anna L David

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Abstract

Uterine artery (UtA) adenovirus vector (Ad)-mediated over-expression of vascular endothelial growth factor (VEGF) enhances uterine blood flow in normal sheep pregnancy and increases fetal growth in the overnourished adolescent sheep model of fetal growth restriction (FGR). Herein we examined its impact on gestation length, neonatal survival, early postnatal growth and metabolism. Singleton-bearing ewes were evenly allocated to receive Ad.VEGF-A165 (5 x 1010 particles/ml, 10 ml, n =17) or Saline (10 ml, n = 16) injected into each UtA at laparotomy (0.6 gestation). Fetal growth was serially monitored (blind) by ultrasound until delivery. Lambs were weighed and blood-sampled weekly and a glucose tolerance test performed (68d postnatal age). Hepatic DNA/RNA was extracted at necropsy (83d postnatal age) to examine methylation status of eight somatotropic axis genes. IGF1 mRNA and protein expression were measured by RT-PCR and radioimmunoassay, respectively. All pregnancies remained viable following Ad.VEGF-A165 treatment. Fetal abdominal circumference and renal volume were greater in Ad.VEGF-A165 versus Saline groups at 21/28 days (p ≤ 0.04) post-injection. At delivery, gestation length (p = 0.07), lamb birthweight (p = 0.08), umbilical girth (p = 0.06) and plasma glucose (p=0.09) tended to be greater in Ad.VEGF-A165 treated lambs. Levels of neonatal intervention required to ensure survival was equivalent between groups. Absolute postnatal growth rate (p = 0.02), insulin area-under-the-curve (p = 0.04) and carcass weight at necropsy (p = 0.04) were increased by Ad.VEGF-A165 treatment. There was no impact on markers of insulin sensitivity or methylation/expression of key genes involved in somatic growth. Ad.VEGF-A165 gene therapy increased fetal growth in a sheep FGR model and lambs continued to thrive during the neonatal and early postnatal period.
Original languageEnglish
Article number142
JournalBiology of Reproduction
Volume94
Issue number6
Early online date20 Apr 2016
DOIs
Publication statusPublished - 1 Jun 2016

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Genetic Therapy
Vascular Endothelial Growth Factor A
Sheep
Fetal Development
Growth
Pregnancy
Uterine Artery
Methylation
Umbilicus
Glucose Tolerance Test
Adenoviridae
Laparotomy
Area Under Curve
Radioimmunoassay
Insulin Resistance
RNA
Insulin
Kidney
Gene Expression
Weights and Measures

Keywords

  • Intrauterine growth restriction (IUGR)
  • gene therapy
  • metabolism
  • sheep
  • vascular endothelial growth factor

Cite this

Peri- and postnatal effects of prenatal adenoviral VEGF gene therapy in growth-restricted sheep. / Carr, David J; Wallace, Jacqueline M; Aitken, Raymond P; Milne, John S; Martin, John F.; Zachary, Ian C.; Peebles, Donald M.; David, Anna L.

In: Biology of Reproduction, Vol. 94, No. 6, 142, 01.06.2016.

Research output: Contribution to journalArticle

Carr, David J ; Wallace, Jacqueline M ; Aitken, Raymond P ; Milne, John S ; Martin, John F. ; Zachary, Ian C. ; Peebles, Donald M. ; David, Anna L. / Peri- and postnatal effects of prenatal adenoviral VEGF gene therapy in growth-restricted sheep. In: Biology of Reproduction. 2016 ; Vol. 94, No. 6.
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N2 - Uterine artery (UtA) adenovirus vector (Ad)-mediated over-expression of vascular endothelial growth factor (VEGF) enhances uterine blood flow in normal sheep pregnancy and increases fetal growth in the overnourished adolescent sheep model of fetal growth restriction (FGR). Herein we examined its impact on gestation length, neonatal survival, early postnatal growth and metabolism. Singleton-bearing ewes were evenly allocated to receive Ad.VEGF-A165 (5 x 1010 particles/ml, 10 ml, n =17) or Saline (10 ml, n = 16) injected into each UtA at laparotomy (0.6 gestation). Fetal growth was serially monitored (blind) by ultrasound until delivery. Lambs were weighed and blood-sampled weekly and a glucose tolerance test performed (68d postnatal age). Hepatic DNA/RNA was extracted at necropsy (83d postnatal age) to examine methylation status of eight somatotropic axis genes. IGF1 mRNA and protein expression were measured by RT-PCR and radioimmunoassay, respectively. All pregnancies remained viable following Ad.VEGF-A165 treatment. Fetal abdominal circumference and renal volume were greater in Ad.VEGF-A165 versus Saline groups at 21/28 days (p ≤ 0.04) post-injection. At delivery, gestation length (p = 0.07), lamb birthweight (p = 0.08), umbilical girth (p = 0.06) and plasma glucose (p=0.09) tended to be greater in Ad.VEGF-A165 treated lambs. Levels of neonatal intervention required to ensure survival was equivalent between groups. Absolute postnatal growth rate (p = 0.02), insulin area-under-the-curve (p = 0.04) and carcass weight at necropsy (p = 0.04) were increased by Ad.VEGF-A165 treatment. There was no impact on markers of insulin sensitivity or methylation/expression of key genes involved in somatic growth. Ad.VEGF-A165 gene therapy increased fetal growth in a sheep FGR model and lambs continued to thrive during the neonatal and early postnatal period.

AB - Uterine artery (UtA) adenovirus vector (Ad)-mediated over-expression of vascular endothelial growth factor (VEGF) enhances uterine blood flow in normal sheep pregnancy and increases fetal growth in the overnourished adolescent sheep model of fetal growth restriction (FGR). Herein we examined its impact on gestation length, neonatal survival, early postnatal growth and metabolism. Singleton-bearing ewes were evenly allocated to receive Ad.VEGF-A165 (5 x 1010 particles/ml, 10 ml, n =17) or Saline (10 ml, n = 16) injected into each UtA at laparotomy (0.6 gestation). Fetal growth was serially monitored (blind) by ultrasound until delivery. Lambs were weighed and blood-sampled weekly and a glucose tolerance test performed (68d postnatal age). Hepatic DNA/RNA was extracted at necropsy (83d postnatal age) to examine methylation status of eight somatotropic axis genes. IGF1 mRNA and protein expression were measured by RT-PCR and radioimmunoassay, respectively. All pregnancies remained viable following Ad.VEGF-A165 treatment. Fetal abdominal circumference and renal volume were greater in Ad.VEGF-A165 versus Saline groups at 21/28 days (p ≤ 0.04) post-injection. At delivery, gestation length (p = 0.07), lamb birthweight (p = 0.08), umbilical girth (p = 0.06) and plasma glucose (p=0.09) tended to be greater in Ad.VEGF-A165 treated lambs. Levels of neonatal intervention required to ensure survival was equivalent between groups. Absolute postnatal growth rate (p = 0.02), insulin area-under-the-curve (p = 0.04) and carcass weight at necropsy (p = 0.04) were increased by Ad.VEGF-A165 treatment. There was no impact on markers of insulin sensitivity or methylation/expression of key genes involved in somatic growth. Ad.VEGF-A165 gene therapy increased fetal growth in a sheep FGR model and lambs continued to thrive during the neonatal and early postnatal period.

KW - Intrauterine growth restriction (IUGR)

KW - gene therapy

KW - metabolism

KW - sheep

KW - vascular endothelial growth factor

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DO - 10.1095/biolreprod.115.133744

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VL - 94

JO - Biology of Reproduction

JF - Biology of Reproduction

SN - 0006-3363

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ER -