PONE-D-21-18174

Exploring the role of Epidermal Growth Factor Receptor Variant III in meningeal tumors

PLOS ONE

Dear Dr. Yadav,

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Academic Editor

PLOS ONE

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'This study was supported by the Sir Ganga Ram Hospital (SGRH) Delhi, India and funded by

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LSRB/81/48222/LSRB-369/PEE&BS/2020).

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Additional Editor Comments (if provided):

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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: Yes

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: 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

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: Yes

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 study attempts the confer searches for the position attained by EGFR vIII in progression and expression of meningioma. Immunohistochemistry technique showed that EGFR vIII is highly expressed in benign tumors as compared to the atypical meningioma with a highly significant p-value (p<0.05). The study id good to explore EGFR vIII in Meningioma. In my opinion paper need to minor revision.

1. In page 4, reference no. 10 (Ge at al., 2002), explaining ligand independent activation of EGFR vIII should be elaborated. Further studies could be carried out on this aspect of the mutant.

2. Page 5, line no. 92 there is a grammatical mistake. ‘Levels’ should be in singular form i.e. ‘level’.

3. Method for Ki-67 analysis should be added in the material and method section.

4. Details of primary antibody of EGFR vIII used for immunohistochemistry need to be provided.

5. In the section of DNA analysis, melting temperature (Tm) of primers used should be mentioned.

6. Details of Chemidoc and the software used for the analysis of blots are required to be given.

7. There is a typing error in detail of Fig. 1, ‘her’ should be written as ‘here’.

8. There is a grammatical error in page 5, line no. 100, DNA sequences should be written as ‘DNA sequence’.

Reviewer #2: Rana et. al. reported EGFR vIII in progression and expression of meningioma. . Hence, Opinions regarding the role that EGFR vIII in tumorigenesis and tumor progression are clearly conflicting and, therefore, it is crucial not only to find out its mechanism of action, but also to definitely identify its role in meningioma. This study need to minor revision for publication.

1. Please elaborate the correlation of gender with EGFR vIII in your study.

2. Discuss about the effect of age on the neurological deficit factor of meningioma.

3. In the section material and methods, RT- PCR parameters used should be added.

4. As mentioned in the text, the tissue samples were stored in -80 degree Celsius. Can you elaborate the extent of degradation in the samples?

5. I would suggest the addition of protocol followed for quantitative PCR.

6. Discuss about the duration of exposure with antibodies in western blotting.

7. Under the section results, line 351-352, pg. no. 20 has some grammatical error.

Reviewer #3: Author reported potentialities of the variant in number of tumors but cannot validate the results by conventional methods and further advanced techniques could be helpful in this regard. Moreover, it can also be said that EGFR vIII does not have any significant role in meningioma. This study need minor correction for before publication.

1. In your study, emphasis is given on Ki-67 proliferation. Its significance in this study should be discussed thoroughly.

2. Authors should provide their perspective for non-significant results by different techniques used.

3. Case recurring meningiomas should be discussed by the authors.

4. In the section material and methods, provide the concentration of formalin used for fixation of tissues.

5. Page 23, line 414, ‘targeting’ should be written as ‘targeted’.

6. Under the section discussion, cross-reactivity of antibodies should be discussed briefly.

7. Discuss the future prospects of this study.

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Reviewer #1: Yes: Abhishek Kumar Singh

Reviewer #2: Yes: Yogesh Kumar

Reviewer #3: No

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Exploring the role of Epidermal Growth Factor Receptor Variant III in meningeal tumors

PONE-D-21-18174R1

Dear Dr. Yadav,

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,

Timir Tripathi, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The reviewers have reviewed the revised manuscript. The authors managed to revise the manuscript as per suggestions and made appropriate changes. Also, they were able to reply to the queries adequately. 

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

**********

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

Reviewer #1: Yes

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 #1: Yes

Reviewer #2: 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

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 #1: Authors have revised the manuscript adequately. The manuscript may be accepted for publication in its current form.

Reviewer #2: All the comments has been addressed properly in the revised manuscript. I recommend this manuscript for publication.

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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: Yes: Yogesh Kumar