Reciprocal Dysregulation of MiR-146b and MiR-451 Contributes in Malignant Phenotype of Follicular Thyroid Tumor

Round 1

Reviewer 1 Report

The authors, Margarita Knyazeva et al., of the submitted original manuscript “Reciprocal dysregulation of miR-146b and miR-451 contributes in malignant phenotype of follicular thyroid tumor” have conducted an exhaustive study with great potential in the clinic. Liquid or conventional biopsies are a powerful tool in the early diagnosis of tumor processes, and within the different molecules that can be analyzed in these biopsies, miRNAs are key, since specific miRNAs signatures are key to unravel the tumor process both in diagnosis as in its evolution. However, as the authors well point out, miRNAs due to their intrinsic characteristics (especially their small size) are difficult to analyze, and a major problem is the lack of a standard normalizer. The methodology proposed by the authors, from my humble point of view, seems to me a very interesting and revealing starting point for the use of miRNAs as a tool, since it is well known that in vitro studies or with few patients show the importance of miRNAs that later does not turn out to be "so good" in the clinic, and it is because cancer is a very heterogeneous and multifactorial disease, so it is extremely difficult for a single molecule to offer too much information, that is why the combination of several miRNAs offered by the authors seems to me a firm step towards a very valuable tool.

In addition to the potential described, it is also important to note that the work is well developed, well presented and that both the methodology and the results, figures and tables are suitable for publication.

For all these reasons, I consider it to be a suitable manuscript for publication with the text as it currently stands. The authors only have to review the affiliations because I have detected some errors, and the nomenclature of the figures, because figure “4A” appears in the text but the figure is not subdivided into panels, and figure 5 should be referenced not only in the discussion, if not also in the results.

Author Response

Dear Reviewer, 

thank you very much for careful consideration of our manuscript! 

All mentioned mistakes are corrected including errors in affiliations and subdivision of Figure 4. Figure 5 (miR-146b/miR-451 induced expression alterations) is now mentioned in the Result section. 

With best results and on  behalf of all authors,

Malek A

Reviewer 2 Report

This is well-designed study to apply method by Androivic (ref 22) to the precious sample fro follicular tumor of thyroid. The problems in this area raised by the authors in the introduction are understandable.

The study first screen the several miRNAs using pooled samples, then individual ones. Even using this quantitation of miRNA may have some overlapped area of the values among three categories, and this grayness reflects reality as whole in Fig 3. 

1. Several cases (5  in miFTC  and 4 in wiFTC in Fig 3 upper right panel, for example) clearly shows the prominent increase in the quantity. Are these cases available giving the readers more precise  histopathological or prognostic information ?
2. Though the markers such as RAS, PAX8, TP53 have limited value in predicting these category of tumors as stated by the authors, any cases here had this kind of markers?
3. I wonder these two miRNA works in other categories of thyroid tumor. Maybe the author can add the in silico data.

Author Response

Dear Reviewer, 

Thank you very much for careful consideration of our manuscript and questions.

1. Several cases (5  in miFTC  and 4 in wiFTC in Fig 3 upper right panel, for example) clearly shows the prominent increase in the quantity. Are these cases available giving the readers more precise  histopathological or prognostic information?

When specific miRNA expression alterations were assayed, our pathologists revised slides and tried to find any correlation between histopathological and miRNA-expression features. in widely invasive FTC group. However, they didn't come with any conclusion. Group of minimally invasive FTC was not evaluated by such way. So, we did not include this information into manuscript. We also can not say anything about prognosis because we don't yet have follow up information. However, now we are testing routinely expression of several marker miRNA in all operated thyroid nodules and we will definitely address this question as soon as will have sufficient amount of samples. 

2. Though the markers such as RAS, PAX8, TP53 have limited value in predicting these category of tumors as stated by the authors, any cases here had this kind of markers?

RAS status has been tested in some cases of FTC but not in all. Therefore we did not included this information in analysis. However, some link between mutations of well-known oncogenes and miRNA expression alterations can be considered and investigated in future. 

I wonder these two miRNA works in other categories of thyroid tumor. Maybe the author can add the in silico data.

It's exciting question! Actually our analysis included also other groups of thyroid cancer (papillary, follicular variant of PTC and poorly differentiated FTC) and we detected involvement of these two (miRNA-146b and miR-451) and other miRNAs. However, we received most prominent preliminary results for groups of samples with follicular nodules first and decided to address this issue in details and to publish these results. So, results of similar analysis of other types of thyroid cancer will be ready soon as separate article. 

With best regards and on behalf of all authors, 

Malek A.