PONE-D-19-23967

Two Novel Mutations in MSX1 Causing Oligodontia

PLOS ONE

Dear Dr. Bian,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process by two reviewer's comments and suggestions.

This editor has additional suggestions:

1. In abstract: page 2, line 12: it seems the “with” should be replaced with “between”;
2. In abstract: page 2, line 22: “ultimately” can be replaced with “thereby potentially”;
3.  In the Introduction part, 2^nd paragraph (dealing with the MSX1 gene) could be moved to Discussion part.  Instead, briefly introducing signaling and genetic contributions of each of nine genes (mentioned in the 1^st paragraph) to tooth agenesis.
4. In the Introduction part, 3rd paragraph: please give reasons why the five genes (MSX1, et al.) are selected to screen the affected individuals.  
5. In Results part, page 8, line 141: please add the range of homeodomain in amino acid residue numbers (from amino acid residual 172 to 234?).
6. Based on current version of HGMD, the number of different missense mutations in the HOX domain should be at least 12, not 7.
7. Page 10, line 186-196, figure 3C-3D of this part can be included as a supplementary figure.
8. In Discussion part, page 11, line 205-206, replace “demonstrate” with “show” and “leads” with “may lead”.
9. Page 11, line 211-212, the statement in 2016 may not be true any longer.  Please check current professional version of HGMD.
10. Page 11, line 215-220, again, the number missense mutations in the HOX domain is more than 7. Please check current professional version of HGMD, and try to include all missense alleles.
11. Figure 4 can be used as supplementary figure.
12. Suggestion: Please consider to have a phenotypic comparison of all missense-related in non-HOX domain to the Table 3 results. If it turns out to be significant, Table 2 as a summary can be added to the main-text.

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Kind regards,

Tao Cai, M.D., Ph.D.

Academic Editor

PLOS ONE

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Comments to the Author

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

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Reviewer #1: Yes

Reviewer #2: Partly

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

Reviewer #1: N/A

Reviewer #2: No

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. 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: 1. In this study, authors identified the MSX1mutations responsible for the tooth agenesis in two Chinese patients, and explained the reasons why these mutations could lead to tooth agenesis by 3D-structure modeling. On the other hand, the authors reviewed all known MSX1 missense mutations available as that are believed to cause non-syndromic tooth agenesis. So I suggest that the title should be changed to “Two Novel Mutations in MSX1 Causing Oligodontia: Case Report and Literature Review”.

2. A frameshift mutation c.590_594 dup TGTCC was detected in family 2. In order to be more accurate, cloning and sequencing should be used to detect the frameshift mutation in III:1 and II:2 in family 2.

Reviewer #2: The manuscript by Yang et al reported two novel MSX1 mutations responsible for non-syndromic tooth agenesis. The authors also reviewed genotype-phenotype relationship of MSX1 missense mutations via 3D-structural analysis and concluded that MSX1 mutations that altered hydrogen bonding tend to cause a more severe non-syndromic TA phenotype with more missing teeth. The analysis is based on 7 mutations identified in the published literature. Overall the study is interesting and well designed. However, there are some issues that need to be addressed before publication.

1. The author concluded that MSX1 mutations that altered hydrogen bonding tend to cause a more severe non-syndromic TA phenotype with more missing teeth. How did they get this conclusion? Because there was not any statistical genotype-phenotype correlation analysis compared between the MSX1 mutations with or without altered hydrogen bonding. What is the average number of missing teeth in MSX1 mutations with or without altered hydrogen bonding respectively? How many MSX1 mutations without altered hydrogen bonding were included for the statistical analysis and the reason for selection should be well described.

2. The manuscript needs to be carefully revised and well organized. In the Discussion part, from line 211 to line 282, include the descriptions of methods, results and discussion about MSX1 mutations that altered hydrogen bonding seems to cause more severe non-syndromic TA phenotype.

3. All the figures are of poor quality, not clear. They should provide better and higher resolution images to readers.

4. Mutational analysis was only performed in the proband of family 2. The pathogenic mutations of other members in this family are needed to be confirmed, especially his mother (II:2)，since she was also a tooth agenesis patient.

5. How did they get the results of Figure 5 should be well descripted, because that is the key result supporting their conclusion. It should be include in the main body of the manuscript.

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Reviewer #1: No

Reviewer #2: No

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Two Novel Mutations in MSX1 Causing Oligodontia

PONE-D-19-23967R1

Dear Dr. Bian,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

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With kind regards,

Tao Cai, M.D., Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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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 #2: Yes

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

Reviewer #2: Yes

**********

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 #2: Yes

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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 #2: Yes

**********

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 #2: The authors have adequately addressed your comments raised in a previous round of review, there is no further comments. The manuscript can be accepted now.

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Reviewer #2: No