PONE-D-20-28747

Mutation status and prognostic value of KRAS and NRAS mutations in Moroccan colon cancer patients: a first report.

PLOS ONE

Dear Dr. Fatima

 The reviewers raised major concerns including: How MIS of CRC established, why an unmethylated control used in Pyrosequencing, since the study is not about methylation?

there different NRAS mutations levels. It should be consistence. 

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Reviewers' comments:

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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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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: No

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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 paper describes RAS mutations in Moroccan CRC patients (210). While the data is sound and confirms previous findings from other populations, this is the first of its kind in Morrocan patients. Changes need to be made to improve the manuscript. The Abstract is too long and the Discussion needs some revision.

Why is it that an unmethylated control used in Purosequencing, this study is not about methylation?

The authors need to explain how was MSI status established, It is NOT i Methods section?

In Methods , they stated they compared stges I-II vs II-IV, I think you mean I-II vs. III-IV, Pease correct.

What is most important is Positive and examined Lymph nodes, NOT harvested/Collected LN. Please adjust write up to show that mutant Kras generally needed LESS LN to be examined to get positve ones.

For NRAS mutations , is it 2-8 or 2-9%, you are putting different numbers in different places?

Would suggest to drop RAS muataions part from text and directly just state KRAS and NRAS.

I tables, please bolden and underline SIGNIFICANT p VALUES.

The paper also uses many unusual abbreviations amd has many typos and grammatical errors. Please review completely.

Reviewer #2: There is a slight redundancy in the Introduction such as “Ras-family G-proteins transduce signals in growth factor signals, such as EGF receptor (EGFR) signaling pathway (1). I suggest you strike out “signals” that appears before “in growth factor”.

The citation numbers need a single space separation from the words. Make space between the last words before cited reference numbers in your text.

Your statement that “KRAS mutations occur in 45% of colorectal cancer (CRC) and NRAS in only 5-8%(6)” should be clarified to indicate whether this is applicable to the Moroccan patients or is referring to the world-wide CRC patients.

-Essentially, the authors of this article are attempting to mimic the work of the French authors in Molecular Cancer Genetics Platform of Poitiers (French), who have published their results about the clinicopathological characteristics of the RAS mutations in a cohort of over 1500 colorectal cancer patients. Was the French study done exclusively with French CRC patients?

-Table 6. Any reason why the multivariate analysis was not done in some of the metastatic tumors in Table 6, where the molecular and clinical variables associated with overall survivals in the 210 colon cancer patients were shown?

-In the body of the text, especially in the Results section, it was quite difficult to follow the different mutational hotspots from gene site to gene site, it would have been better to display the most common types in tabular format for quicker views and understanding of the cardinal gene alterations.

-The impetus to investigate the WT and mutated RAS gene sites was that mutations in KRAS outside exon 2 (KRAS exons 3,4) and in other downstream effectors of the EGFR signaling pathway (like NRAS) were published to be associated with resistance to anti-EGFR monoclonal antibodies that were used as a standard regimen for CRC patients – meaning that the two Anti-EGFR monoclonal antibodies, panitumumab and cetuximab, used to treat CRC patients with KRAS and NRAS mutations, were not effective. Therefore, the policy was to successfully treat CRC patients, the full RAS WT status had to be confirmed in all mCRC patients before initiating treatment. The question is: Does this study conclusively resolve that issue? If so, that must be clearly stated in the manuscript.

-Compared to NRAS wild-type patients, “NRAS-mutated patients seem to be exhibiting lower total lymph node rate, and were associated with higher lymph node metastasis number”. This statement is confusing. It could be revised to make the right statement without seemingly contradicting itself. Among RAS mutations, KRAS codon 13 mutations seemed to have a poor prognostic value, which may negatively impinge on overall survival of all the stages of CRC patients. Is that a correct statement?

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Reviewer #2: No

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Mutation status and prognostic value of KRAS and NRAS mutations in Moroccan colon cancer patients: a first report.

PONE-D-20-28747R1

Dear Dr. FATIMA EL AGY

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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Hassan Ashktorab

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 #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

**********

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: No

**********

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: This is a much improved manuscript. However, the English grammar issue still persist throughout the manuscript and especially in the Introduction section stating with the first sentence where "in" before the word "transduce" must be taken out. Also, somewhere further down the paragraphs "gander" was used for "gender..

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Reviewer #2: No