CHILDHOOD OBESITY IN ALABAMA: RISK FACTORS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1017405
• Project Start Date: Oct 5, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Recipient Organization
AUBURN UNIVERSITY
108 M. WHITE SMITH HALL
AUBURN,AL 36849

Performing Department
Nutrition and Food Science

Non Technical Summary
Childhood obesity is a major public health concern in the United States. Obese children are at higher risk to develop type 2 diabetes, cardiovascular disease, cancer, sleep apnea and a greater risk of social and psychological problems. In the United States, approximately 12.7 million children and adolescents are classified as obese (1).The prevalence of obesity is 8.9% among 2 to 5 year-olds compared with 17.5% of 6 to 11 year-olds and 20.5% of 12 to 19 year-olds (2). Around $190 billion dollars have been estimated as annual medical expenditures associated with obesity.The long-term goal is to improve the health of children specifically in Alabama by identifying the multiple factors, such as biological, genetic, lifestyle, socioeconomic, dietary habits, and parental influences that could lead to childhood obesity. This will help to identify appropriate interventions to reduce childhood obesity and related health outcomes in minority children.The overarching hypothesis of this proposal is that identification of multiple factors that could lead to childhood obesity, such as behavioral, lifestyle, socioeconomic, dietary habits, parental influences, genetic, and biological factors, will help in the development and implementation of innovative tailored solutions to reduce obesity.

Animal Health Component

0%

Research Effort Categories

Basic

100%

Applied

0%

Developmental

0%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
724	5010	1060	50%
801	6099	1080	50%

Knowledge Area
724 - Healthy Lifestyle; 801 - Individual and Family Resource Management;

Subject Of Investigation
6099 - People and communities, general/other; 5010 - Food;

Field Of Science
1060 - Biology (whole systems); 1080 - Genetics;

Keywords

childhood obesity

alabama

behavioral factors

salivary amylase

microbiome

Goals / Objectives
The Specific objectives are:1. To validate the relationship between extents of physical activity, eating, sleeping habits and family influences to the development of childhood obesity, we will develop and use questionnaire-based estimation to identify the socioeconomic, anthropometric, behavioral factors, child's eating habits, physical activity and extent of parental misperception of child weight status.2. To determine the genetic factors involved in the childhood obesity, the genomic copy number variants (CNVs) of salivary a-amylase will be determined in relation to BMI.3. To investigate the differences in the composition of salivary and gut microbiome in lean, overweight and obese children, as the relative species abundance and metabolic characteristics of the salivary and gut microbiome may change substantially in obese compared to healthy children.

Project Methods
ParticipantsFor this research we will be recruiting around 200 children ages 6 to 10 years from Lee and Macon county of Alabama. Each participant will be paid $50 cash as appreciation of their participation. The study has been approved by Institutional Review Board of Auburn University.Inclusion Criteria: 1) 6 to 10 year's old; 2) White/European or African American ethnicity. Exclusion Criteria: 1) Type 1 or 2 diabetes or major health issues; 2) Taking antibiotics.Parents and children will be requested to sign the written consent. Height and weight of the participating children will be measured, without shoes and wearing only light clothing using standard protocol; height will be measured to the nearest 1/8 inch stadiometer, and weight will be measured to the nearest 4 ounces using a Tanita digital scale (53). The weight and height will be used to calculate the BMI, to assess body fat and approximate the ideal weight for a person, considering the sex and age. The children will be classified as obese, overweight, normal weight and underweight. Since children grow with age and not all such growth is body fat, their BMI is compared with age and sex related percentiles within the ages of 2 to 20. The Center for Disease Control and Prevention (CDC) standard for BMI in children is as follows: "Underweight: Less than the 5th percentile; Healthy weight: 5th percentile to less than the 85th percentile; Overweight: 85th to less than the 95th percentile; Obese: Equal to or greater than the 95th percentile." The biological samples such as saliva and fecal samples will be collected from all the participants. The parents were asked to fill out a few questionnaires to gather data about:The lifestyle factors that can lead to increased risk for obesity, such as time spent watching television/electronics, dinner schedule, amount of sleep and physical activity. The maternal education and family's income will also be recorded to determine the socioeconomic factors.The Child Eating Behavior Questionnaire (CEBQ) questionnaire to evaluate eight subscales of eating behavior: Food Responsiveness, Enjoyment of Food, Emotional Over-Eating, Desire to Drink, Slowness in Eating, Satiety Responsiveness, Food Fussiness and Emotional Under-Eating (54).The aspect of family environment, parental beliefs, attitudes and parents child-feeding practices, like restriction and pressure to eat will be determined by using the Child Feeding questionnaire (CFQ) (55, 56).Salivary a-amylase Copy Number VariantsSaliva will be collected from the participants using the DNA GenoTek Saliva Collection Kit. Genomic DNA from the saliva will be extracted by the PrepIT.L2P method (DNA GenoTek, Ontario, Canada), according to manufacturer's protocol. Copy number of the AMY1 gene will be estimated by a duplex quantitative real-time PCR (QuantStudio™ 3 qPCR) containing two TaqMan assays (Life Technologies), one for AMY1 (Hs07226361_cn, FAM-labeled) and one specific for the reference gene (RNase P, VIC labeled). Relative copy number values will be calculated by the ΔΔCT method with the Hap-Map sample NA18956 as calibrator. Linear regression analyses will then be performed to assess the association between AMY1 copy number and BMI.Fecal Sample 16S Sequencing and MetabolomicsMicrobial DNA will be isolated from fecal samples using the MoBio Power Soil Kit. In brief, a fecal sample is added to a bead beating tube and subjected to methodical homogenization. Cells are lysed by mechanical and chemical methods. From this sample, total genomic DNA is captured on a silica membrane in a spin column format. DNA is then washed, eluted from the membrane and quality determined by a NanoDrop. DNA sequencing of the 16S rRNA gene is amplified by PCR analysis using cDNA and 0.3 μM V3-V5 barcoded primers and other downstream applications. Equal volume of sample will be pooled, then sequenced on one lane of MiSeq using the MiSeq Reagent Kit v2 (Illumina Inc.) (57).Salivary Microbiome AnalysisSalivary samples are collected and centrifuged at 4°C and 1500xg for 15 min and pellet collected and preserved in -80°C for further microbiome analysis. Salivary DNA extraction will be performed using QIAamp DNA blood mini kit. DNA yield and purity will be measured by spectrophotometrically using Nanodrop. PCR will be performed and sequenced by Illumina MiSeq (58).Bioinformatics ProcessingAnalysis of 16S rRNA data requires the use of bioinformatics tools. Using primary default parameters, these tools are used to compare taxonomic classification, diversity analysis, and usability. Open source tools are available such as Quantitative Insights into Microbial Ecology (QIIME)/MetaGenome Rapid Annotation using subsystem Technology (MG-RAST)/Mothur etc. Chimera detection and checking is done using 16s rRNA reference database (59). In QIIME, chimera filtering and quality control is performed by using standards less than 250bp length, containing more than 8 ambiguous bases and average quality score of less than 25. QIIME is compatible with most of the sequencing methods and will be used to perform taxonomic analysis and diversity analysis (60, 61).Statistical AnalysisAll the statistical analysis of the data collected will be conducted by the services of Dr. Philippe Gaillard, Associate Professor, Department of Mathematics and Statistics at the College of Science and Mathematics (COSAM) Statistical Center Services (SCC). Statistical analysis will be performed using SAS version 9.4 (SAS Inc, Carry, North Carolina, USA). Standard and compositional descriptive statistics will be used for comparison. Linear regression models will be used to determine the associations between obesity markers and behavioral factors.

Progress 10/01/20 to 09/30/21

Outputs
Target Audience:The target audience includes undergraduate students, graduate students, faculty, research scientists, and physicians. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Dr. Geetha Thangiah (PI) of this project contributed to the planning and execution. PI also contributed to the experimental design and analysis, manuscript preparation, and revision. The graduate students, undergraduate students, and postdoctoral fellows contributed to the laboratory experiments, analysis of the questionnaire, and statistical analysis. Most of them were also involved in the writing of the manuscript and presentation of the results in local and national meetings. How have the results been disseminated to communities of interest?The findings from this research proposal were reached to the scientific communities through our publications and presentations in local and professional meetings. What do you plan to do during the next reporting period to accomplish the goals?We are planning to investigate the genetic factors involved in childhood obesity, the genomic copy number variants in lean, overweight, and obese children.

Impacts
What was accomplished under these goals? The overall goal of this proposed study is to evaluate the multiple factors such as environmental, lifestyle, economic, parental influences, biological, genetic, oral, and gut microbiomes that could lead to childhood obesity. Objective 1: Determine the relationship of the behavioral factors associated with childhood obesity. Findings: We determined the association between maternal education and eating behavior in children. The increased food responsiveness and emotional overeating in obese children are influenced by maternal education. Objective 2: Determine the genetic factors associated with childhood obesity. Findings: We explored the connection between telomere length ratio, blood pressure, and childhood obesity. The overweight/obese children have higher systolic pressure compared to normal-weight children. African American children have a greater mean telomere length ratio and blood pressure compared to European American children. Objective 3: Determine the salivary and gut microbiome in childhood obesity. Findings: We attempted to generate a profile of gut and oral microbial clades predictive of disease status in African American and European American children. Gut and oral microbial diversity between African American and European American children showed significant differences in alpha-, beta-, and taxa-level diversity. Socioeconomic factors influenced microbiota in obesity, and it was ethnicity-dependent.

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Ayine P, Selvaraju V, Venkatapoorna CMK, Bao Y, Gaillard P, Geetha T. Eating behaviors in relation to child weight status and maternal education. Children 2021 Jan 7;8(1):32. doi: 10.3390/children8010032.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Selvaraju V, Phillips M, Fouty A, Babu JR, Geetha T. Telomere length as a biomarker for race-related health disparities. Genes 2021 Jan 9;12(1):78. doi: 10.3390/genes12010078.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Balakrishnan B, Selvaraju V, Chen J, Ayine P, Yang L, Babu JR, Geetha T, Taneja V. Ethnic variability associating gut and oral microbiome with obesity in children. Gut Microbes 2021 Jan-Dec;13(1):1-15. doi: 10.1080/19490976.2021.1882926.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Selvaraju V, Babu JR, Geetha T. Telomere length as a biomarker for race-related health disparities. Health Disparities Research Initiative, Auburn University, Auburn, AL, USA, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Ayine P, Selvaraju V, Venkatapoorna CMK, Bao Y, Gaillard P, Geetha T. The Childrens Eating Behaviors: Relation to child weight status and maternal education. CHS Graduate Student Research Symposium, Auburn University, Auburn, AL, USA, March 19, 2021.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Patel P, Selvaraju V, Geetha T. Epigenetics, diet, and obesity. CHS Graduate Student Research Symposium, Auburn University, Auburn, AL, USA, March 19, 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Ayine P, Selvaraju V, Venkatapoorna CMK, Bao Y, Gaillard P, Geetha T. The childrens eating behaviors: Relation to child weight status and maternal education. Auburn Research Student Symposium, Auburn University, Auburn, AL, USA, March 29, 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Chester B, Selvaraju V, Martin K, Geetha T. Evaluating the Relationship Between Obesity and the Efficacy of a Diabetes Self-Management Education and Support Coaching Structure for the Ongoing Management of Type 2 Diabetes in North Alabama. Boshell Diabetes & Metabolic Diseases, 13th Annual Research Day, Auburn University, Auburn, AL, USA, September 17, 2021. Abstract # O04.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Selvaraju V, Balakrishnan B, Chen J, Ayine P, Yang L, Babu JR, Taneja V, Geetha T. Ethnic-specific Association of Gut Microbiome with Childhood Obesity. Boshell Diabetes & Metabolic Diseases, 13th Annual Research Day, Auburn University, Auburn, AL, USA, September 17, 2021. Abstract # P32.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Patel P, Selvaraju V, Babu JR, Taneja V, Geetha T. Epigenetic Changes in NRF1 and FTO Gene Through MethyLight Assay Amongst Races in Children. Boshell Diabetes & Metabolic Diseases, 13th Annual Research Day, Auburn University, Auburn, AL, USA, September 17, 2021. Abstract # P20.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:The target audience include undergraduate students, graduate students, faculty, research scientists, and physicians. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Dr. Geetha Thangiah (PI) of this project contributed to the planning and execution. PI also contributed to the experimental design and analysis, manuscript preparation and revision. The graduate students, undergraduate students, and postdoctoral fellows contributed to the laboratory experiments, analysis of the questionnaire and statistical analysis. Most of them were also involved in the writing the manuscript and presentation of the results in local and national meetings. How have the results been disseminated to communities of interest?The findings from this research proposal were reached to the scientific communities through our publications and presentations in local and professional meetings. What do you plan to do during the next reporting period to accomplish the goals?We are planning to investigate the differences in the composition of salivary and gut microbiome in lean, overweight and obese children.

Impacts
What was accomplished under these goals? The overall goal of this proposed study is to evaluate the multiple factors such as environmental, lifestyle, economic, parental influences, biological, genetic, oral and gut microbiomes that could lead to childhood obesity Objective 1: Determine the relationship of the behavioral factors associated with childhood obesity. Findings: 1.Around 71% of children with late bedtime (8:48 pm), 75% of children who watch television for more than 1 hour, and 54% of children who have dinner after 7:00 pm have obesity. Daily behavior habits such as late bedtime, increased television exposure, and late dinnertime are associated with obesity. 2.The BMI z-score was significantly associated with parental perceived child weight and concern about child weight. The BMI z-score had a significant negative association with parents pressuring children to eat. Only the parental feeding practice pressure to eat was dependent upon the maternal education. Objective 2: Determine the genetic factors associated with childhood obesity. Findings: 1. The mean AMY1 copy number significantly decreased in overweight/obese compared to normal weight children. AMY1 copy number inversely associated with the obesity measurements. 2. The salivary amylase gene copy number is associated with the inflammatory markers in children. These findings were published in peer-reviewed manuscripts and presented in state and local meetings.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Selvaraju V, Ayine P, Parra EP, Fadamiro M, Babu JR, Brown M, Geetha T. Urinary biomarkers of inflammation and oxidative stress are elevated in obese children and correlate with a marker of endothelial dysfunction. Oxid Med Cell Longev. 2019 Oct 16;2019:9604740. doi: 10.1155/2019/9604740.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Venkatapoorna CMK, Ayine P, Selvaraju V, Parra EP, Koenigs T, Babu JR, Geetha T. The relationship between obesity and sleep timing behavior, television exposure, and dinnertime among elementary school-age children. J Clin Sleep Med. 2020 Jan 15;16(1):129-136. doi: 10.5664/jcsm.8080.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Ayine P, Selvaraju V, Venkatapoorna CMK, Geetha T. Parental feeding practices in relation to maternal education and childhood obesity. Nutrients 2020 Apr 9;12(4):1033. doi: 10.3390/nu12041033.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Phillips M, Babu JR, Wang X, Geetha T. DNA copy number and structural variation (CNV) contributions to adult and childhood obesity. Biochem Soc Trans. 2020 Aug 28;48(4):1819-1828. doi: 10.1042/BST20200556.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Phillips M, Selvaraju V, Fouty A, Sandey M, Jeganathan R, Geetha T. Gender differences in the copy number of 11q11 gene in European American obese children. Diabetes 2020; 66: 1256-P.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Selvaraju V, Phillips M, Fouty A, Babu JR, Geetha T. Telomere length as a biomarker for race-related health disparities. Health Disparities Research Initiative Virtual Symposium, Auburn University, Auburn, AL, USA, July 29, 2020.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Ayine P, Selvaraju V, Geetha T. Parental feeding practices and perceptions of child weight in relation to maternal education and childhood obesity. Virtual Symposium, Auburn University, Auburn, AL, USA, April 2020.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Fouty A, Blocker A, Phillips M, Selvaraju V, Babu JR, Sandey M, Geetha T. The relationship between the copy number of 11q11 gene and childhood obesity. Virtual Symposium, Auburn University, Auburn, AL, USA, April 2020.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Selvaraju V, Babu JR, Geetha T. Association of Salivary neurotrophins and insulin in childhood obesity. Center for Neuroscience Initiative Inaugural Retreat, Auburn University, Auburn, AL, USA, February 28, 2020. Abstract No. 57.

Progress 10/05/18 to 09/30/19

Outputs
Target Audience:Scientific communities including undergraduate students, graduate students, postdoctoral fellows, faculty, and research scientists. Changes/Problems:None What opportunities for training and professional development has the project provided?Dr. Geetha Thangiah (PI) of this project contributed to the planning and execution. PI also contributed to the experimental design and analysis, manuscript preparation and revision. The graduate students, undergraduate students, and postdoctoral fellows contributed to the recruitment of the participants, collection of anthropometric measurements and biological samples, laboratory experiments, and analysis. Most of them were also involved in the presentation of the results in local and national meetings. How have the results been disseminated to communities of interest?The findings from this research proposal were reached to the scientific communities through our publications and presentations in local and professional meetings. What do you plan to do during the next reporting period to accomplish the goals?We are planning to study the behavioral factors such as daily lifestyle habits, parental perceptions, and eating behaviorassociated with childhood obesity.

Impacts
What was accomplished under these goals? We recruited 169 participants aged 6 to 10 years from Lee and Macon County. We determined the relationship between the copy number of salivary amylase (AMY1) gene and obesity measurements in children. In addition, we examined the level of the salivary obesity-related biomarkers in normal weight and overweight/obese children, association with the obesity measures and the interrelations between the biomarkers.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Venkatapoorna CMK, Ayine P, Parra EP, Koenigs T, Phillips M, Babu JR, Sandey M, Geetha T. Association of salivary amylase (AMY1) gene copy number with obesity in Alabama elementary school children. Nutrients 2019 Jun 19;11(6). doi: 10.3390/nu11061379
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Selvaraju V, Babu JR, Geetha T. Association of salivary C-reactive protein with the obesity measures and markers in children. Diabetes Metab Syndr Obes. 2019 12:1239-1247. doi: 10.2147/DMSO.S211624.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Sycheva M, Sustarich J, Zhang Y, Selvaraju V, Geetha T, Gearing M, Babu JR. Pro-Nerve Growth Factor Induces Activation of RhoA Kinase and Neuronal Cell Death. Brain Sci. 2019 9(8): 204. doi: 10.3390/brainsci9080204.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Vines K, Li R, Geetha T, Broderick TL, Carroll CC, Babu JR. Nerve growth factor receptor TrkA signaling in streptozotocin-induced type 1 diabetes rat brain. Biochem Biophys Res Commun. 2019 514(4):1285-1289. doi: 10.1016/j.bbrc.2019.04.162.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Chester B, Babu JR, Greene MW, Geetha T. The effects of popular diets on type 2 diabetes management. Diabetes Metab Res Rev. 2019 35(8):e3188. doi: 10.1002/dmrr.3188.
• Type: Conference Papers and Presentations Status: Other Year Published: 2018 Citation: Geetha T, Venkatapoorna C, Ayine P, Parra EP, Koenigs T, Babu JR. Salivary amylase and childhood obesity. 2018 AAES Faculty Summit Auburn University, Auburn, AL, USA, Dec 10, 2018.
• Type: Conference Papers and Presentations Status: Other Year Published: 2019 Citation: Venkatapoorna C, Ayine P, Parra EP, Koenigs T, Sandey M, Babu JR, Geetha T. Low AMY1 gene copy number Is associated with Increased childhood obesity. 12th Annual Boshell Diabetes and Metabolic Diseases conference, Auburn University, Auburn, AL, USA, February 15, 2019. Abstract No. O14.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Selvaraju V, Ayine P, Parra EP, Brown M, Babu JR, Geetha T. Association of inflammation, oxidative stress and endothelial dysfunction biomarkers in obese elementary school children. 12th Annual Boshell Diabetes and Metabolic Diseases conference, Auburn University, Auburn, AL, USA, February 15, 2019. Abstract No. P31.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Ayine P, Venkatapoorna C, Parra EP, Koenigs T, Selvaraju V, Babu JR, Geetha T. Exploring the relationship between sleep, dinner and television timing behavior with BMI in children. 12th Annual Boshell Diabetes and Metabolic Diseases conference, Auburn University, Auburn, AL, USA, February 15, 2019. Abstract No. P06.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Koenigs T, Venkatapoorna C, Ayine P, Parra EP, Sandey M, Babu JR, Geetha T. The salivary amylase gene copy number and childhood obesity. 2019 ALDA Annual meeting and Food EXPO, Renaissance Montgomery Hotel & Spa, Montgomery, AL, USA, March 6, 2019. Abstract No. P06.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Parra EP, Ayine P, Venkatapoorna C, Koenigs T, Selvaraju V, Babu JR, Geetha T. The relationship between sleep behavior with obesity, television exposure and dinner time in children. 2019 Auburn Research Student Symposium, Auburn University, Auburn, AL, USA, April 9, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Koenigs T, Venkatapoorna C, Ayine P, Parra EP, Sandey M, Babu JR, Geetha T. Reduced AMY1 gene copy number Is associated with Increased BMI in children. 2019 Auburn Research Student Symposium, Auburn University, Auburn, AL, USA, April 9, 2019.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Venkatapoorna C, Ayine P, Parra EP, Sandey M, Babu JR, Geetha T. Low AMY1 Gene Copy Number Is Associated with Increased Childhood Obesity in Alabama. American diabetes Association 79th Scientific Sessions June 7-11, 2019, San Francisco, CA. Abstract # 2053-P.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Selvaraju V, Ayine P, Parra EP, Brown M, Babu JR, Geetha T. Endothelial Dysfunction, Inflammation, and Oxidative Stress Urinary Biomarkers in Obese Children from Alabama. American diabetes Association 79th Scientific Sessions June 7-11, 2019, San Francisco, CA. Abstract # 2054-P.