PONE-D-20-14717

Hemopexin and α1-microglobulin heme scavengers with differential involvement in preeclampsia and fetal growth restriction

PLOS ONE

Dear Dr. Youssef,

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.

SPECIFIC ACADEMIC EDITOR COMMENTS: There were two expert reviewers in the field that handled your manuscript. We thank them for their time. Although interest was found in your study, several major points arose during review that require your attention. Comments that need addressing include: 1) several questions about the methods, 2) the need for correlative studies, and 3) measures of oxidative stress. Please address all of the reviewers' comments in your revised manuscript.

Please submit your revised manuscript by Aug 03 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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We look forward to receiving your revised manuscript.

Kind regards,

Frank T. Spradley

Academic Editor

PLOS ONE

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3. Thank you for submitting the above manuscript to PLOS ONE. During our internal evaluation of the manuscript, we found some minor text overlap between your Discussion section (limitations) of your submission and the following previously published work:

https://www.ajog.org/article/S0002-9378(19)30909-3/fulltext

Please revise the manuscript to rephrase the duplicated text and ensure you cite your sources.

4. Please report the source, catalog number, and dilutions of all antibodies used in your study. Please also report the catalog number of all commercial kits and immunoassays used.

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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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

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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: 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: The study presented by Youssef et al. evaluated maternal and fetal plasma

concentrations of heme scavengers (Hemopexin and α1-microglobulin) in different phenotypes of placenta-mediated disorders and controls, both in mothers and neonates.

To prevent the toxic, and notably, the pro-oxidant effect of free hemoglobin and its degradation compounds haem and free iron, the human body disposes of circulating scavenger proteins, including notably those two molecules, that have been associated with preeclampsia.

The study includes 47 singleton pregnancies collected in a monocentric prospective study.

Its originality is related to the exploration of those two haem scavengers both mothers and neonates groups in a same study with the same methods, compared to previously published study.

The manuscript is overall well written and clear. The population is well described, the mention of potential treatments for PE should be cited.

The limits of the paper are clearly exposed.

The most surprising result is the difference observed for A1 microglobulin between mothers and fetuses, in contrast with hemopexin. This point is debated in the discussion section.

Main corrections

A flow chart of the study cohort should be presented.

The quality of the figure and legend should be increased. The legend has to permit the reader to understand them without the text. What type of graph is presented? Median IQ? A line showing which populations have been compared with p should be easier to read.

The analysis of the quality of the homemade A1M assay should be presented briefly. Notably, the standard curve should be presented in supporting pieces of information, or a previous publication should be cited, if it do exist. The manufacturer’s providing the monoclonal antibodies should be mentioned. Whether all samples were analyzed in a single batch and if some internal quality controls were included should be precised (since unknown samples are cited).

As the renal function is supposed to explain the increase of A1M in mothers, an analysis of A1M concentration regarding the creatinine concentration would possibly reinforce this hypothesis. Some additional explanations should be explored, such as albumin and haptoglobin concentrations in maternal and fetal bloods.

Minor corrections

The population is extremely well described. However, some data could be suppressed because there are discussed further. For instance, the classification between mild and severe is not use in the analysis of the data, so I’m not sure it deserves to be mentioned in this setting.

It should be mentioned whether the assays were done in singlicates for hemopexin.

Units have to be corrected ug/mL replace by �g/mL using symbol font.

The legend of the supporting tables has to be clarified: I think only mean + Sd is presented for biological values, whereas we can read “Data are mean (Å} standard deviation) or median (interquartile range).”

Line 131 : three instead of 3

Line 132 : two instead of 2

Reviewer #2: Overall, well performed set of experiments. I have only minor comments.

1. There's obviously a wide range of lab values in each group. Was any attempt made to correlate the levels of A1M or hemopexin with overall outcomes? Time to delivery, maternal morbidity, et cetera.

2. Was any attempt made to measure markers of oxidative stress? Oxidized lipids, for instance?

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Reviewer #1: No

Reviewer #2: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PONE-D-20-14717R1

Hemopexin and α1-microglobulin heme scavengers with differential involvement in preeclampsia and fetal growth restriction

PLOS ONE

Dear Dr. Youssef,

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.

ACADEMIC EDITOR COMMENTS: There are some remaining comments that must be addressed by the authors in your revised manuscript.

Please submit your revised manuscript by Oct 09 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Frank T. Spradley

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

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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

**********

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

Reviewer #1: 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 #1: Yes

**********

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

**********

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: Thank you for this revised version, that is now for me perfectly acceptable for publication.

I just notice the following mistake to correct :

Legend Fig 1 ; FGR, fetal grown restriction : replace by fetal growth restriction

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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.

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: Yes: Katell Peoc'h

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Hemopexin and α1-microglobulin heme scavengers with differential involvement in preeclampsia and fetal growth restriction

PONE-D-20-14717R2

Dear Dr. Youssef,

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.

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Kind regards,

Frank T. Spradley

Academic Editor

PLOS ONE