PONE-D-24-10611Evaluating soluble Axl as a biomarker for glioblastoma: a pilot studyPLOS ONE

Dear Dr. Rovin,

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Academic Editor

PLOS ONE

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Additional Editor Comments:

A number of issues were identified by both reviewers; particularly Reviewer #2. These should be addressed if the authors plan to submit a revised manuscript to the journal.

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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

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

Reviewer #1: No

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: No

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 pilot study conducted by Raymond et al. aimed to assess the potential of soluble Axl (sAxl) as a biomarker candidate for glioblastoma (GBM). Previous research has indicated frequent expression of the AXL receptor and its putative ligand, Gas6, in human gliomas. Notably, both Axl and Gas6 are often overexpressed in glioma and vascular cells, correlating with a poor prognosis in GBM patients (Hutterer et al., reference 14). However, the study found no significant difference in sAXL concentration between GBM patients and a healthy control group.

When examining glioblastoma cell lines, the authors observed significant AXL expression in 45% of the samples, consistent with previous findings. Despite this, the study's results suggest that serum sAXL concentration alone may not serve as a reliable diagnostic tool for glioblastoma.

Given that Hutterer et al. reported a higher number of GBM specimens with elevated GAS6 levels, it would be worthwhile for the authors to consider including Gas6 measurements in their sample analysis. Additionally, although glioblastomas release tumoral content into the bloodstream, and certain biomarkers are known to traverse the blood-brain barrier, it's plausible that some molecules may be retained in the cerebrospinal fluid. Therefore, assessing sAXL levels in cerebrospinal fluid samples could provide valuable insights and should be considered by the authors.

Furthermore, expanding Table 3 to incorporate data from additional studies, such as reference 18 and various studies on hepatocellular carcinoma, would enrich the comparative analysis.

10.4143/crt.2019.749

10.18632/oncotarget.17598

10.1002/ijc.29394

Line 95. “1019”.

Line 187, line 188, line 191, line 219. The correlation of between magnitudes should be quantified and included in the results.

The values of the qRT-PCR analysis should be included in the study, not only the bar. The type of magnitude should be included in the figure legend (mean or median, SEM or SD) with the number of measurements performed.

Reviewer #2: In the manuscript “Evaluating soluble Axl as a biomarker for glioblastoma: a pilot study”，The author found that although AXL is highly expressed in many cancers, including glioblastoma, the byproduct of AXL expression, sAXL, cannot serve as a reliable biomarker for diagnosing glioblastoma. This is a carefully done study and the findings are of considerable interest. A few minor revisions are list below.

1. Results – It would be helpful to the reader if the Figures can be improved with high quality and pixels.

2. Tables – The table looks a little bit complicated, just have a try the three-line table.

3. Does this imply that the experimental results may be meaningless as there is no difference in preoperative AXL levels between the control group and the GBM group?

4. By searching the keyword “glioblastoma” and “AXL”, it is not hard to find other articles on the connection between them. Please describe the difference between this manuscript and other articles and point out the innovations.

5. The language and syntax and format need more attention and the authors might benefit from the assistance of an English language editor.

Suggestion：

1. Use high quality pictures.

2. Identify the innovative points of the article.

3. Expand the sample size and re-study to reduce false-negative results.

4. Some of the references cited in the article are outdated.

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Reviewer #1: No

Reviewer #2: No

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Evaluating soluble Axl as a biomarker for glioblastoma: a pilot study

PONE-D-24-10611R1

Dear Dr. Rovin,

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,

Salvatore V Pizzo

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

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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

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 #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: The authors have addressed my comments. I could not access the data in dryad, possibly the data are not yet available?

Reviewer #2: (No Response)

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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: No

Reviewer #2: No

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