Metabolic Pathology of Pediatric NAFLD

Understanding the Metabolic Pathology of Pediatric Obesity and NAFLD

Nonalcoholic fatty liver disease (NAFLD) is now the most common liver disease worldwide and affects nearly 40% of obese youth and up to 10% of the general pediatric population. Some features of NAFLD are similar in children and adults, yet fibrosis and inflammation are more common in the portal zone and occur earlier in pediatric NAFLD patients than adults. This portends a rapid progression to end-stage liver disease in early adulthood. For the majority of children with NAFLD, mechanisms driving the origin and rapid progression of disease remain unknown. Thus, there is a critical, unmet need to study the specific underlying patterns of metabolic and molecular changes in the liver underlying the development and progression unique to children with NAFLD.

This proposal will test the hypotheses that children with NAFLD have excess glucose and lipid produced by the liver, that those events are regulated by specific variations in the amount and location of RNAs and proteins in liver, and that the concentration of specific micro-RNAs in the blood can be used as a biomarker for NAFLD in pediatric patients.

Study Overview

• Status: Recruiting
• Conditions: Obesity; Nonalcoholic Steatohepatitis; Nonalcoholic Fatty Liver
• Intervention / Treatment: Other: Oral sugar tolerance test; Other: De novo lipogenesis test; Other: Gluconeogenesis test

Detailed Description

This project uses a cross-sectional design with a single testing period without a formal intervention (e.g., diet, drug, exercise) or natural follow-up period. Participants with nonalcoholic fatty liver disease (NAFLD), and age-matched control groups classified as either obese (Ob control) or normal weight (NW control) will complete all metabolic and descriptive tests, including blood analyses.

The NAFLD group will also have a liver biopsy as part of their standard clinical care; a portion of the biopsy will be used for the research testing. The Ob and NW control groups will not undergo liver biopsy. To provide a set of reference liver samples to compare with the NAFLD group, we will enroll a "liver control" group, consisting of age-matched patients who are scheduled to have a cholecystectomy with liver biopsy or are undergoing liver resection for tumor removal.

• Study Type: Interventional
• Enrollment (Estimated): 100
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Kevin Short, PhD; Phone Number: 43094 405-271-8001; Email: kevin-short@ouhsc.edu

Study Locations

• United States
  • Oklahoma
    • Oklahoma City, Oklahoma, United States, 73104
      • Recruiting
      • University of Oklahoma Health Sciences Center
      • Contact: Kevin Short, PhD; Phone Number: 43094 405-271-8001; Email: kevin-short@ouhsc.edu

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 10 years to 20 years (Child, Adult)
• Accepts Healthy Volunteers: Yes

Study Population

Participants in the NAFLD and Liver control groups will be recruited from the OU Health pediatric clinics. Prospective patients will be identified from clinic records and during clinic visits. Participants in the Ob control and NW control groups will be recruited from the local community.

Description

Inclusion Criteria:

• Age: All participants must be 10.0 to 20.9 years old at the time of enrollment.
• Sex: Male and Female participants are eligible.
• Race/Ethnicity: Participants of all racial/ethnic identities will be recruited.
• Body mass index (BMI): Participants must be either in the normal weight (NW control group) or obese [Ob control, nonalcoholic fatty liver disease (NALFD) groups] range for BMI percentile. BMI percentile will be calculated from age- and sex-specific growth charts for children.
• NAFLD status: The NAFLD group participants will be eligible if they are scheduled for liver biopsy for clinical reasons and their histopathology report confirms a diagnosis of NAFLD. NW control, and Ob control, and Liver control participants must not have diagnosed NAFLD.

Exclusion Criteria:

• Chronic illness: Participants will not be able to participate if they have conditions that are likely to affect metabolic variables (either directly or due to required medications) or result in them being unable to complete the required tests. Such conditions could include, but are not limited to, untreated hypothyroidism or other endocrine disorders, rheumatoid arthritis requiring steroids or limiting mobility, cardiovascular disease, stroke, or cardiac failure, neurological disorders such as multiple sclerosis, cancer, liver diseases other than NAFLD (e.g., Wilson's disease), other organ disorders, or orthopedic conditions that limit physical activity.
• Acute illness: Participants will not be able to participate if they develop acute conditions that are likely to affect metabolic outcomes (either directly or due to required medications) or result in them being unable to participate; e.g., respiratory illness, infectious disease, fever, accident resulting in bone fractures, myocardial infarction, major depression. If such conditions resolve and there are no longer risks or likelihood of adverse effect on the study outcomes, participants may be rescheduled for testing.
• Medications and nutritional supplements: Medications, vitamins, or supplements that have known effects on the primary outcomes will be cause for exclusion. Examples include weight loss medications, glucocorticoids, or experimental medications used to correct a metabolic or hepatic condition. Medications used to control asthma, allergies, anxiety, depression, attention deficit disorder, menstrual cycle, hypothyroidism, gastric reflux, hypertension, and sleep will be allowed. Participants who are taking medications for treatment of acute illness or conditions such as cold, flu, injury, or infection will be rescheduled after they complete their treatment course.
• Pregnancy: Evidence of pregnancy or intent to become pregnant during the study is cause for exclusion.
• Smoking, alcohol abuse, or illicit drug abuse: Participants who smoke or have signs or symptoms of alcohol or substance abuse will be excluded.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: NAFLD; Participants in the pediatric NAFLD clinic	Other: Oral sugar tolerance test; Measurement of glucose and insulin for calculation of insulin sensitivity; Other: De novo lipogenesis test; Oral consumption of deuterated water to measure incorporation of label into lipids; Other: Gluconeogenesis test; Oral consumption of 13C-labeled glycerol to measure incorporation into glucose
Experimental: Ob control; Participants with obesity, without NAFLD	Other: Oral sugar tolerance test; Measurement of glucose and insulin for calculation of insulin sensitivity; Other: De novo lipogenesis test; Oral consumption of deuterated water to measure incorporation of label into lipids; Other: Gluconeogenesis test; Oral consumption of 13C-labeled glycerol to measure incorporation into glucose
Experimental: NW control; Participants in the normal range for body weight, without NAFLD	Other: Oral sugar tolerance test; Measurement of glucose and insulin for calculation of insulin sensitivity; Other: De novo lipogenesis test; Oral consumption of deuterated water to measure incorporation of label into lipids; Other: Gluconeogenesis test; Oral consumption of 13C-labeled glycerol to measure incorporation into glucose
Experimental: Liver control; Participants undergoing liver biopsy or liver surgery, without NAFLD	Other: Oral sugar tolerance test; Measurement of glucose and insulin for calculation of insulin sensitivity; Other: De novo lipogenesis test; Oral consumption of deuterated water to measure incorporation of label into lipids; Other: Gluconeogenesis test; Oral consumption of 13C-labeled glycerol to measure incorporation into glucose

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
De novo lipogenesis	Measurement of the rate of newly synthesized triglycerides in plasma using deuterated water	Day 1
Gluconeogenesis	Measurement of the rate newly synthesized glucose in circulation using labeled glycerol and deuterated water	Day 1
Serum microRNA	Abundance of microRNAs in serum using a broad profiling platform and real-time quantitative polymerase chain reaction tests for confirming individual miRNAs	Day 1
Abundance of liver collagen	Abundance of collagen in liver biopsy sections, using Second Harmonic Generation microscopy	Day 1
Insulin sensitivity	Calculated value of insulin sensitivity, using the oral minimal model and serial concentrations of glucose and insulin during an oral sugar tolerance test	Day 1
Liver mitochondrial flux	Reported as the fluorescent lifetime redox ratio (FLIRR), which is calculated from measurements of free and bound NADH and FAD in liver biopsy sections, using fluorescence lifetime imaging microscopy	Day 1

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Targets of microRNA-122	Transcripts bound to microRNA-122, measured using high-throughput sequencing of cross- linked immunoprecipitates (HITS-CLIP)	Day 1
Liver transcriptomics	Spatial distribution of messenger RNAs in liver biopsies	Day 1

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Blood pressure	Brachial and central blood pressures	Day 1
Arterial stiffness	Measured as carotid-femoral pulse wave velocity	Day 1
Body composition	Whole body and regional lean and fat mass, measure with dual energy X-ray absorptiometry	Day 1
Cardiorespiratory fitness	Peak oxygen consumption during bicycle ergometry test to volitional fatigue	Day 1
Blood DNA analysis	Measurement of single-nucleotide polymorphisms associated with NAFLD risk	Day 1
Liver steatosis	Use of Fibroscan to measure controlled attenuation parameter (measure of steatosis)	Day 1
Liver fibrosis	Use of Fibroscan to measure elastic modulus (surrogate measure of fibrosis)	Day 1

Collaborators and Investigators

• Sponsor: University of Oklahoma
• Collaborators: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• Investigators: Principal Investigator: Kevin Short, PhD, University of Oklahoma

Study record dates

Study Major Dates

• Study Start (Actual): May 25, 2022
• Primary Completion (Estimated): February 1, 2026
• Study Completion (Estimated): June 1, 2026

Study Registration Dates

• First Submitted: June 1, 2022
• First Submitted That Met QC Criteria: June 17, 2022
• First Posted (Actual): June 24, 2022

Study Record Updates

• Last Update Posted (Actual): March 6, 2024
• Last Update Submitted That Met QC Criteria: March 4, 2024
• Last Verified: March 1, 2024

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Digestive System Diseases; Liver Diseases; Overnutrition; Nutrition Disorders; Overweight; Body Weight; Obesity; Fatty Liver; Non-alcoholic Fatty Liver Disease
• Other Study ID Numbers: 14018; 1R01DK129656-01A1 (U.S. NIH Grant/Contract)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No