Prevalence of TERT Promoter Mutations in Orbital Solitary Fibrous Tumors

Round 1

Reviewer 1 Report

It is a good study, but the following points are noteworthy

1.       The keywords are duplicated.

2.       Figure 1 and Figure 2A and 3 are not mentioned in the text

3.       Figure 3 is not clear, it is suggested to send a clear and specific figure

4.       A good study has been done, but it is difficult to draw conclusions with this small number of samples. Is it possible to check more samples?

5.       Many markers are mentioned in the execution method, but they are not mentioned in the results and discussion, such as Bcl2 …..

It is good, needs further investigation

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in yellow.

Comments 1.:The keywords are duplicated.

Response 1.: Thank you for pointing this out. We have corrected this.

Comments 2: Figure 1 and Figure 2A and 3 are not mentioned in the text.

Response 2.: Thank you for mentioning this. We have now taken care that all of of the Figures have an in-text reference (page 5, line 4; page 6, line 3 and 6; page 6, line 9). Please mind that an additional figure has been added and the figure numbering has changed.

Comments 3: Figure 3 is not clear, it is suggested to send a clear and specific figure.

Response 3: Thank you for this comment. We have adjusted the figure's description in order to better explain the importance of histology and immunohistochemistry for diagnosis and prognosis. In the text, we have added a specific description of immunohistochemistry in the results (see page 10, 3.4 pathology, line 11-14 yellow) and in the discussion section (see page 15, 4. Discussion line 21-31 yellow). To make the figure more understandable we have put it together on one single page (page 12).

Comments 4: Thank you for the comment.

Response 4: A multicenter study is in conception.

Comments 5: Many markers are mentioned in the execution method, but they are not mentioned in the results and discussion, such as Bcl2 …..

Response 5: Thank you for pointing this out. We agree with this comment. Therefore we have more precisely explained the immunohistochemistry staining in the result section (see page 10, 3.4 pathology, line 11-14 yellow) and included it in the discussion (see page 15, 4. Discussion line 21-31 yellow) - see also comment 3.

4. Quality of English.

English editing of the text, tables and figure descriptions has been performed by a native speaker.

Reviewer 2 Report

The authors present the article "Prevalence of TERT-promotor mutations in Orbital Solitary Fibrous Tumors", in which they explain the main clinical and molecular characteristics of oSFT in a group of patients.
The authors' proposal is complete and well described; However, I consider that there are several issues that must be improved to make it a more complete description.
The main point has to do with the importance with which they treat TERT promoter mutations.
The topic of TERT promoter mutations is the central axis on which the authors present their paper, since it is part of the title, but during the development of the text, greater importance is given to all the clinical characteristics of the tumors of the patients and very little to mutations, in this regard I have the following comments:
- A better methodological description of TERT promoter sequencing should be made.
- The mutation detected in 1 patient must be better described, for example, it is a heterozygous mutation, etc.
- Is the mutation present only in the samples analyzed or is it present in the germ line?
- What is the frequency of the mutation in the general population?
- Does the mutation affect the binding of any transcription factor that could generate a change in TERT expression?
- In the discussion section, I consider the consequences of the mutations detected in the TERT promoter in the context of TERT expression should be analyzed because the authors propose that the search for mutations at this site should be included in the studies to analyze the behavior of these tumors.
Other comments:

Promoter instead of promotor

The images in Figure 3 should be smaller so that they can be viewed on a single page.
It would be convenient for the authors to include an image of the TERT promoter to better understand its structural characteristics and how mutations at this site can affect the expression of the gene.

In conclusion, they must increase and improve what is related to the description of mutations in the TERT promoter.

Minor editing of English language required

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files.

Comment 1: - A better methodological description of TERT promoter sequencing should be made.

Response 1: Thank you for pointing this out. We have added the methodological description of the TERT promotor sequencing in the M&M section (page 4, 2.5 Analyses of molecular pathology).

Comment 2: - The mutation detected in 1 patient must be better described, for example, it is a heterozygous mutation, etc.
- Is the mutation present only in the samples analyzed or is it present in the germ line?
- What is the frequency of the mutation in the general population?

Response 2: Thank you for the advice. We have referred to the questions in the introduction (page 2, 1. Introduction, line 45-48 and page 3, 1. Introduction, line 1-5) and the discussion section (page 17, 4. Discussion, line 6-8).

Comment 3: - Does the mutation affect the binding of any transcription factor that could generate a change in TERT expression?
- In the discussion section, I consider the consequences of the mutations detected in the TERT promoter in the context of TERT expression should be analyzed because the authors propose that the search for mutations at this site should be included in the studies to analyze the behavior of these tumors.
- In conclusion, they must increase and improve what is related to the description of mutations in the TERT promoter.

Response 3: Thank you for pointing this out. Accordingly, we have given an overview of the pathways of the mutated TERT promotor in the introduction (including an image as suggested in comment 5) (page 2, 1. Introduction, line 45-48 and page 3, 1. Introduction, line 1-5) and highlighted consequences of the mutations on TERT expression in the discussion section (page 16, 4. Discussion, line 38-52 and page 17, 4. Discussion, line 1-5).

Comments 4: Promoter instead of promotor

Response 4: Agree. We have accordingly changed the terminus, including the title.

Comments 5: The images in Figure 3 should be smaller so that they can be viewed on a single page.
It would be convenient for the authors to include an image of the TERT promoter to better understand its structural characteristics and how mutations at this site can affect the expression of the gene.

Response 5: Thank you for pointing this out. We have done the suggested modifications (page 12, 3.4. pathology) and (page 3, 1. Introduction).

4. Response to Comments on the Quality of English Language

Response 1: English editing of the Title, Abstract, Text, Tables and figure description has been done by a native speaker.

Reviewer 3 Report

The manuscript entitled "Prevalence of TERT-promotor mutations in Orbital Solitary Fibrous Tumors" submitted by Koca et.al discusses the use promotor mutations in the TERT gene in the orbital compartment and correlating it with the clinical appearance, histology, and molecular pathology. The study helps to identify the prognostic factors in the oSFT by using the promotor mutations in the TERT gene.

The authors have a clear hypothesis and to test this the author used a retrospective study to identify whether the oSFT has any association with the TRET-promotor mutations. The statistical methods used are appropriate to the best of my knowledge. The discussion is well written. However, there are some concerns as follows.

The material and methods sections need to be organised into sub-headings. 

The authors should mention how did they sequence the TERT-promotor mutations using the samples, the DNA extraction method and the primers used.

The authors need to mention the Figure 1 in the main text of the results section.

Under the discussion section the authors need to cite certain recent mechanisms by which these mutations in the promotors could affect the nearby genes as shown in the article PMID: 38049665 for certain cancers. 

Overall, the work done by Koca et.al. is commendable. 

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files.

Comments 1: The material and methods sections need to be organised into sub-headings. 

Response 1: Thank you for pointing this out. Accordingly, we have modified the M&M section (page 3 and page 4).

Comments 2: The authors should mention how did they sequence the TERT-promotor mutations using the samples, the DNA extraction method and the primers used.

Response 2: Agree. For this, we have added a detailed description of the sanger-sequencing (page 4, 2.5. Analyses of molecular pathology, line 34-43).

Comments 3: The authors need to mention the Figure 1 in the main text of the results section.

Response 3: Thank you for the comment. We agree, accordingly, we have mentioned the figure in the text (page 5, 3.1 Epidemiological and clinical parameters, line 4 and page 6, 3.2 Imaging, line 6). Please take note, that the the numbering of the figures has changed since we have included a new figure 1 as schematic illustration of the mutated TERT promoter (page 3).

Comment 4: Under the discussion section the authors need to cite certain recent mechanisms by which these mutations in the promotors could affect the nearby genes as shown in the article PMID: 38049665 for certain cancers. 

Response 4: Thank you for pointing this out. Accordingly, we have discussed how the two hotspot mutations affect the related genes (page 16, 5. Discussion, line 38-32 and page 17, 5. Discussion, line 1-5). In the introduction section, we have included a description (page 2, 1. Introduction, line 45-48 and page 3, 1. introduction, line 1-5) and schematic illustration (page 3) of the mutated gene sequence to support the discussion section.

Round 2

Reviewer 2 Report

The manuscript could be accepted in present form.