PONE-D-25-62126Genetic variants in the Leptin-Melanocortin Pathway and their joint effects with physical activity and sleep duration on risk of childhood obesityPLOS One

Dear Dr. Liu,

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Academic Editor

PLOS One

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Reviewers' comments:

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: Yes

Reviewer #3: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: No

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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5. Review Comments to the Author

Reviewer #1: The case–control study examines genetic variants in the leptin–melanocortin pathway and their associations with childhood obesity in a Chinese population.

The identification of 4 obesity-associated variants and the GRS analyses are supported by existing literature. The application of CART analysis adds value by identifying 2 variants as important contributors despite their lack of statistical significance in single-locus models.

The study did not detect statistically significant additive or multiplicative gene–lifestyle interactions, despite observing joint effects, which contrasts with some larger studies. The authors acknowledge this limitation and attribute it primarily to limited statistical power. Gene–environment interaction analyses typically require larger samples. The self-reported and dichotomised lifestyle variables likely introduce measurement error and reduce power to detect interactions. The cross-sectional design, combined with a retrospective lifestyle assessment, increases susceptibility to recall bias and reverse causation.

Although BMI is considered sufficient for the diagnosis of obesity in children, clinicians should be aware of its limitations. Also, precise, standardised measurement of weight and height using calibrated equipment and proper technique is essential. How did you do “twice averaged”? Please comment.

The non-continuous age range may affect generalizability. Confounding from unmeasured factors such as diet, screen time, and parental obesity remains possible. The restriction to Han Chinese children from Guangzhou further limits generalizability.

The study reinforces the importance of the leptin–melanocortin pathway in childhood obesity. The finding that children with both a high genetic burden and adverse lifestyle factors experience substantially elevated obesity risk has potential relevance for risk stratification and prevention strategies. Modest effect sizes and the cross-sectional design limit clinical translation.

I would suggest clarifying the limitations of interaction analyses more explicitly and avoiding overinterpretation of joint effects as evidence of interaction. I would consider the value of the study primarily in genetic association and GRS findings rather than gene–environment interaction claims.

I recommend that the manuscript be accepted after minor revisions.

Reviewer #2: The study aimed to explore gene-lifestyle interactions between functional genetic variants in leptin melanocortin pathway and obesity in childhood. The authors have constructed genetic risk scores and categorized children into low, medium and high-risk groups for development of obesity based on four important genetic variants and explored the interaction in relation to sleep and exercise habits. The study is very interesting and addresses important modifiable risk factors, which brings the novelty and applicability of the study. However, some sections would benefit with minor revisions.

1. In the introduction section, please consider providing greater detail and mechanism of the proposed interaction between sleep and physical exercise on the expression in the leptin melanocortin pathway (Paragraph 83-90 or in Figure 1).

Methodology is very robust and explained in detail. Results are well presented. Moderate sample size limited the possibility of finding statistically significant interactions between high genetic risk scores and inadequate physical activity and sleep duration, which was emphasized in the discussion.

2. In the discussion section, I suggest an explanation on the lack of significant differences in physical activity and sleep duration between cases and controls (paragraph 195-199).

The Conclusion section is concise and addresses future directions.

Reviewer #3: The authors have conducted and written a thorough study on the effect of genetic risk scores derived from variants on the leptin-melanocortin pathway and their effects on childhood obesity, as well as the interaction between the genetic risk scores with some lifestyle behaviors (sleep and physical activity) on the risk of obesity. The study strengths include a frequency-matched case control design, rigorous statistics and a robust sample size. They genotyped twelve potentially functional variants and identified four of them as being associated with childhood obesity, and constructed a GRS using those variants. They found that a high unweighted and weighted GRS using those variants conferred a high risk of childhood obesity, with a joint effect from poor physical activity or sleep.

My main comment pertains to whether the authors may be able to explain at least briefly in the manuscript why they decided to focus specifically on physical activity and sleep and why other lifestyle factors such as dietary quality for example were not included.

My second comment pertains to data availability. Since the authors selected that some restrictions apply, would they be able to describe in detail what these restrictions are?

Minor issues were a few small grammatical errors found throughout the text, would recommend a thorough check for these (examples: line 252, line 314, line 319).

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

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Attachments

Attachment

Submitted filename: GeneticVariants_review.docx

Genetic variants in the Leptin-Melanocortin Pathway and their joint effects with physical activity and sleep duration on risk of childhood obesity

PONE-D-25-62126R1

Dear Dr. Li Liu,

we re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. Attention has to be paid to the data availability upon reasonable request.

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Kind regards,

Nataša Marčun Varda, PhD

Academic Editor

PLOS One

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: No

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #2: Comment on Q4: Individual-level raw data are not publicly available due to participant privacy considerations, ethical restrictions, and the Regulation on the Administration of Human Genetic Resources of China. Data may be available upon reasonable request via email.

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If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

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