PONE-D-20-26604

Serum soluble B7-H3 is a prognostic marker for patients with non-muscle-invasive bladder cancer.

PLOS ONE

Dear Dr. Azuma,

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PLOS ONE

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2. We noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed:

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0199719 (Introduction paragraph 3)

https://www.jimmunol.org/content/185/6/3677.full (Discussion paragraph 5)

https://www.nature.com/articles/srep27528 (Discussion paragraph 7)

https://www.clinical-genitourinary-cancer.com/article/S1558-7673(13)00049-9/fulltext (Discussion paragraph 7)

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed.

3. Please provide a brief summary of the methods regarding the detection and measurement of sB7-H3 in serum. This is in line with our reproducibility criterion for publishing, see https://journals.plos.org/plosone/s/criteria-for-publication.

4. In your Methods section, please provide additional information about the collection of control tissue specimens used in this study, the method used to collect them, and the demographic details of the patients from which they were collected. Please ensure you have provided sufficient details to replicate the analyses such as: a) the  date range (month and year) during which you collected specimens,  b) a description of how participants were recruited to provide samples, and c) eligibility criteria for being included in this part of the study.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. 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: Partly

Reviewer #3: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

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3. 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: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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4. 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: No

Reviewer #2: Yes

Reviewer #3: 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: This manuscript is addressing a very interesting question asking if B7-H4, a specific member of the immunstimulatory B7 family, in its soluble from can be used as a prognostic marker in NMIBC.

A strength of this project is its large patient cohort.

However, there are several aspects in the manuscript that should be improved.

1) the introduction is not written in a precise and clear style and order.

2) The authors should carefully ensure that they really describe what they are doing. (for example under the headline "presence of serum sB7-H3 in patients with NMIBC" they start with "This assay revealed that..." without mentioning which assay they mean).

3) There is a very recent paper from Podojil et al., on B7-H4 in urothelial carcinoma that should be included into the discussion ( PMID: 32363111 ).

Reviewer #2: This is a review of a historical series of 555 patients with NMIBC, in which sB7-H3 has been measured in plasma, relating it to progression and recurrence-free survival.

The authors should be congratulated for an exhaustive work, as well as the description of a promising marker in the study of recurrence and progression of NMIBC. Anyway, I think the work may improve with the following contributions:

Introduction: The first paragraph seems to be missing. If not, it is advisable to check the grammar.

Material and methods: Given that the primary objective is recurrence, it should be indicated how these patients have been followed (cystoscopy, cytology, ultrasound, CT ...). In the definitions part, progression is defined as “beyond T2 stage”. I suppose they mean "beyond T1 stage", to diagnose progression to muscle infiltrating.

It does not show how these patients were treated. Given that most are intermediate or high risk tumours, the logical thing is that they had some treatment. The therapy applied logically may have influenced the recurrence, altering the results of the study. If this data is not available, it should be included among the weaknesses of the work, in the discussion section.

Results: The median follow-up of the entire series is missing, not only of the tumors that have recurred, as well as whether there were any lost to follow-up.

At the bottom of Figure 2, the results of the curves are discussed. This data is already given in the text, so duplication should be avoided.

As there are some healthy donors positive for sB7H3, can a cut-off level be defined to know when it is considered positive?

Given that tumors with positive sB7H3 are usually recurrent, I recommend doing uni / multivariate analysis of the variable “prior recurrence”, grouping primary vs recurrent. It is possible that it may be significant.

Discussion

The analysis has been carried out on a series for which the treatment applied is not available. If the percentage of patients who have received BCG is high, the EORTC tables loses validity, making it more advisable to use the risk according to CUETO (Babjuck et al, EAU Guidelines NMIBC 2020). If the information is available, it is advisable to see if these results hold up. If so, the importance of this marker would increase, as it would be significant even when a more “modern” BCG treatment is performed. And it can serve as an indicator to predict response to BCG, something much needed today.

Conclusions

Nothing to add

Reviewer #3: The manuscript “Serum soluble B7-H3 is a prognostic marker for patients with non-muscle-invasive bladder cancer“ by et al address an important point in NMIBC that is the finding of good prognostic marker for recurrence and progression of the disease. The has evaluated 555 patient samples and found a correlation between the serum levels of B7-H3 and disease recurrence and progression to MIBC. The article is well written but I miss some aspect that I think need to be elucidated.

1.- The authors point the NMIBC need of good markers for disease progression, but they not make a reasoning of why to measure B7-H3, is there any rationale for that or is just because?

2.- With the patient descriptions it is not clear how many of the patients are primary or recurrent in each analysis. It will be interesting to have the B7-H3 values dissected by patients’ groups not just by positive vs negative. Is there a cut value that really distinguish healthy vs cancer patient?

3.-Currently the patients that are considered high risk of tumor progression according to EORT guidelines are treated with intravesical instillations. Is that the case for some of those patients in the cohort?. In the same line, it will be interesting to know how are the B7-H3 serum levels in a cystitis or a bladder infection patient. Will B7-H3 be as HD or as a cancer patient?

4.- I miss the analysis of how varied (or not) the B7-H3 serum levels among the patients with a primary tumor vs those that are recurrent, not only in positive vs negative but the quantitative amount.

5.- In table 2 which is the unit of the tumor size? Cm?. The authors distinguish Ta T1 and Tis, how many of the patients have Tis plus Ta or T1 and how do this affect the analysis?

6.- The authors point in mat. and meth. that the samples were collected between 2008 and 2013. I thnk it will be important to know the follow up time.

7.- In page 16, last paragraph, the author wrote that in the samples they found an 8% of tumor progression (47 samples) but in figure 4 they make the Kaplan analysis with the 555 samples. I think they should make this among the samples of recurrent patients not with the total.

8.- And last, I think that the authors most do an effort to point clearly how many of the patients that are not categorized now of high risk of recurrence or progression will be benefit with the measurement of sB7-H3. To me it is not clear as it is now.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

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Serum soluble B7-H3 is a prognostic marker for patients with non-muscle-invasive bladder cancer.

PONE-D-20-26604R1

Dear Dr. Azuma,

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.

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Kind regards,

Francisco X. Real

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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

Reviewer #3: 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: (No Response)

Reviewer #2: (No Response)

Reviewer #3: Yes

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

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: 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 Response)

Reviewer #3: 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 #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: 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: (No Response)

Reviewer #3: The authors addressed most of the comments raised the previous review. I think that this manuscript is now acceptable for publication. Congratulations!!

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No