Necrotizing Enterocolitis, Prematurity
Conditions
Keywords
newborn, necrotizing enterocolitis, genetics, growth factors, metabolism
Brief summary
This proposal will test the hypothesis that synthesis and catabolism of epidermal growth factor (EGF), the genotype of the EGF gene, and the microbiome interact to influence EGF expression in infants at risk for necrotizing enterocolitis (NEC).
Detailed description
* Preterm infants will receive a six hour intravenous infusion of \[5,5,5-2H3\]leucine (2H3) through an existing intravenous line (IV) to measure EGF synthesis rate. * Two blood samples will be obtained, one prior to the start of infusion, and one during the infusion. The enrichment of the stable isotope labeled leucine will be measured in the plasma from these samples; DNA will be extracted from the residual cell pellets. The EGF and EGF receptor genes will be sequenced. * Saliva and urine will be obtained for 5 days following infusion to measure EGF and the rate of incorporation of leucine into EGF using liquid chromatography (LC)/mass spectroscopy (MS)/MS technology, as well as enzyme-linked immunosorbent assay (ELISA) . Saliva will be obtained by a Q tip swab and urine and stool obtained from the diaper. * Stool will be obtained every 3 to 7 days through 5 weeks to evaluate inflammatory markers and the microbiome. * If breastfeeding, a single sample of mother's milk will be obtained for measurement of EGF after adequate volumes for infant feeds are achieved.
Interventions
BIOLOGICAL[5,5,5-2H3]leucine (stable isotope labeled leucine)
intravenous infusion of labeled leucine dissolved in 5% glucose water: priming dose of 18 micromoles (1.8 ml)/kg over 5 minutes, then 18 micromoles (1.8 ml)/hr for 6 hours; one infusion total
Sponsors
Washington University School of Medicine
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
BASIC_SCIENCE
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
1 Days to 7 Days
Healthy volunteers
No
Inclusion criteria
* gestation 32 weeks or less * 1 week of age or less * intravenous line in place for clinical purposes
Exclusion criteria
* imminent death * active infection * pre-existing diagnosis of NEC * fluid or electrolyte imbalance
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Salivary EGF (Epidermal Growth Factor) Protein Levels | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | Salivary EGF protein levels obtained from oral swabs were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to micrograms of protein in saliva, and expressed as picogram of EGF protein per microgram of total salivary protein. |
| Urinary EGF Protein Levels | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | Urinary EGF protein levels obtained from free flowing urine samples retrieved from subject diaper were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to milligrams of creatinine in urine, and expressed as nanograms of EGF protein per milligram of urinary creatinine. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| EGF Gene Sequencing | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | Identification of computationally predicted functional variants in EGF gene |
Countries
United States
Participant flow
Recruitment details
Subjects admitted to the neonatal intensive care unit at St. Louis Children's Hospital were screened and enrolled from 5/1/2008 to 12/31/2009. Infants born at \< 32 weeks and 7 days old or less were eligible for enrollment which occurred in the neonatal intensive care unit or postpartum floor.
Pre-assignment details
After enrollment 11 of 29 enrolled subjects did not complete the study due to removal of the intravenous line required for infusion, significant deterioration in clinical status resulting in unstable condition as identified by the attending physician, parental withdrawal, and death.
Participants by arm
| Arm | Count |
|---|---|
| Preterm Infants for EGF Profiles Premature infants born at \< 32 weeks gestation who are 7 days old or less. Infants received and intravenous infusion of \[5,5,5-2H3\]leucine (stable isotope labeled leucine) with sampling of blood, urine and saliva. | 18 |
| Total | 18 |
Withdrawals & dropouts
| Period | Reason | FG000 |
|---|---|---|
| Overall Study | Death | 2 |
| Overall Study | Deterioration in clinical status | 1 |
| Overall Study | Lack of intravenous line | 7 |
| Overall Study | Withdrawal by Subject | 1 |
Baseline characteristics
| Characteristic | Preterm Infants for EGF Profiles |
|---|---|
| Age, Categorical <=18 years | 18 Participants |
| Age, Categorical >=65 years | 0 Participants |
| Age, Categorical Between 18 and 65 years | 0 Participants |
| Age, Continuous | 4 Days |
| Region of Enrollment United States | 18 Participants |
| Sex: Female, Male Female | 6 Participants |
| Sex: Female, Male Male | 12 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | 1 / 29 |
| other Total, other adverse events | 0 / 29 |
| serious Total, serious adverse events | 0 / 29 |
Outcome results
Primary
Salivary EGF (Epidermal Growth Factor) Protein Levels
Salivary EGF protein levels obtained from oral swabs were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to micrograms of protein in saliva, and expressed as picogram of EGF protein per microgram of total salivary protein.
Time frame: Sampling occurred on average day of life 9 with a range from day of life 7 to 21
Population: 29 subjects were enrolled with 11 not completing the study and not analyzed due to lack of specimens
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Preterm Infants for EGF Profiles | Salivary EGF (Epidermal Growth Factor) Protein Levels | 15 picograms EGF/ microgram of protein |
Primary
Urinary EGF Protein Levels
Urinary EGF protein levels obtained from free flowing urine samples retrieved from subject diaper were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to milligrams of creatinine in urine, and expressed as nanograms of EGF protein per milligram of urinary creatinine.
Time frame: Sampling occurred on average day of life 9 with a range from day of life 7 to 21
Population: 29 subjects were enrolled; 11 did not complete the study and were not analyzed
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Preterm Infants for EGF Profiles | Urinary EGF Protein Levels | 48 nanograms of EGF/mg of creatinine |
Secondary
EGF Gene Sequencing
Identification of computationally predicted functional variants in EGF gene
Time frame: Sampling occurred on average day of life 9 with a range from day of life 7 to 21
Population: 29 subjects were enrolled; 11 did not complete the study and were not analyzed
| Arm | Measure | Value (NUMBER) |
|---|---|---|
| Preterm Infants for EGF Profiles | EGF Gene Sequencing | 11 percentage of participants with variant |