CUX1—Transcriptional Master Regulator of Tumor Progression in Pancreatic Neuroendocrine Tumors
, Julia Weissbach 1,†, Annika Blank 2, Aurel Perren 2, Johannes Haybaeck 3,4, Volker Fendrich 5, Anja Rinke 6, Thomas Mathias Gress 6, Jonas Rosendahl 1 and Patrick Michl 1,*
Round 1
Reviewer 1 Report
Thank you for your contribution to this underinvestigated area--I found your paper to be engaging and interesting.
I have a few questions and concerns:
in Results 4.2, it is unclear to me where the specimens are coming from--are these biopsies? Resected specimens? FNAs? I ask because the Ki-67% is exceedingly difficult to predict from aspirates and even cores in non-resected tumors. Also in this result, why is Ki67 ≥ 10% used? This is not a standard cutoff typically for any grading system. Was mitotic index considered?
You note that the WHO 2010 grading schema is used, however you report (in results 4.1) grade 1 vs grade 2/3 tumors (grade 3 NETs were only introduced in the WHO 2017 schema), so perhaps this could be clarified. I would like to see a breakdown of the grades, LN positivity, metastases of these resected tumors as I think it makes a difference in what kind of population of resected specimens you are working with and analyzing.
In line 141 of the review copy, you note that in 37 patients, metastases were available but then characterize these as LN specimens. I think clarification is required as to whether these were lymph node metastases or if any were metastases resected from the liver or otherwise, as in PNETs (unlike the majority of other tumors) liver metastasectomy is performed with some frequency.
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The authors extend their previous observations on the CUX1 transcriptional repressor to non-functional pancreatic NET behavior. They note some indications that higher immunohistochemically detected values of this marker may correlate with reduced progression free survival in surgically treated NET patients. Such significance is noted when binary comparison (IHC score of <8 vs. >8) is applied (Fig. 1C). They also extend these studies to BON1 and QGP1 cell lines and RiPTag2-derived tumors. The latter studies, however, provide a less than clear picture on the negative impact of CUX1 on caspase 3.
The studies are of potential interest. However, they can be improved significantly as follows:
- Statistical analyses using univariate parameters require testing in a multivariate model. This would greatly enhance the clarity of impact of CUX1 vs. recognized clinicopathological parameters in correlating with disease outcomes.\
- Please show the QGP1 data along side the BON1 studies.
- Please show measures of apoptosis throughout. The western blots require PARP degradation products.
- Similarly, the mouse model requires TUNEL/Annexin V staining as a measure of apoptosis to validate the putative function of CUX1.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
The authors have made a reasonable attempt at responding to the questions and concerns raised earlier.
