Review Reports

Round 1

Reviewer 1 Report

Dear authors,

The introduction and the Material and Methods sections are written very well in clarity and references. However, I miss in the Results section completely the finding of NUP98 and Sanger results of these NUP98 mutations, which only pop up in the final stage of your Discussion and are marked as a novel biomarker for recurrent cancer, although this is even not validated by Sanger sequencing as confirmed mutations in the data processing of Figure 3, Fig S2 and even missing in S1 primer list? Further, Supplementary Figures 2 and 3 are low in resolution quality. It's a pity since I read a lot of good analysis results for the primary and secondary tumors, but missing the bridge of the NUP98 as a novel biomarker?

Regards,

Reviewer

Author Response

Dear Reviewer,Thank you for your comment.Here is our reply.

1.  We missed validation of NUP98 After our verification, the mutation of the NUP98 gene was unreliable. We remove all of results about NUP98 gene in our study.
2. We had provided high resolution quality figures in website.
3. We had removed all of results about NUP98 gene in our study.

Best regard,

Jan-Gowth Chang

Reviewer 2 Report

This is an interesting study about mutation analysis of second primary tumors in head and neck cancer in Taiwanese patients through next-generation sequencing. Data from 15 patients with oral recurrent cancer were analyzed.

The paper is well written. However, some issues remain.

Since the authors included only oral tumors, this should be stated in the title.

I think that patients with first or second primary tumors not in the head and neck must be excluded from the study to avoid biases. Indeed, one patient had an ureteral cancer that was not squamous cell carcinoma.

Please add the version of TNM staging system that was used.

The table 1 was not included in the text.

Clinical data about first primary tumors must be added.

Were all the tumors HPV-negative?

The authors stated that three patients in the first group (PA50, PA53, and PA55, 3/9) had recurrent cancer because they did not observe a unique driver gene mutation in the ST. But what about clinical features of such tumors (distance from first and second tumor)?

How can the authors say that one patient in the third group had recurrent cancer if the last was in a different organ compared to the first primary?

Please report how much time was between first and second tumor.

Author Response

Dear reviewer,Thank you for your comment

1.  We had removed two patients with ureteral and lung cancer (PA48 and PA51).
2. The table 1 include clinical data (distance, TNM staging system, and time was between first and second tumor). HPV infection test is not a routine test in clinical guidelines in CMUH, so HPV were not included in our data.