PONE-D-19-33979

Cancer cell-derived interleukin-33 decoy receptor sST2 enhances orthotopic tumor growth in a murine pancreatic cancer model

PLOS ONE

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Reviewers' comments:

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Comments to the Author

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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: Yes

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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: IL-33 has been widely studied in inflammatory diseases, but its role in cancer is somewhat paradoxical. Human pancreatic cancer is one of the deadliest malignancies wherein the inflammation of the pancreas plays a critical role. This research paper entitled “Cancer cell-derived interleukin-33 decoy receptor sST2 enhances orthotopic tumor growth in a murine pancreatic cancer model” encompasses some interesting findings wherein following issues should be addressed.

Minor Concerns:

1. Fig. 1 legend can be made more explanatory.

2. Fig. 4 legend need correction wherein last line saying *p<0.005 should be corrected as *p<0.05.

Major Concerns:

1. Fig.1: Authors should provide a plot for Δct values for ST2l and sST2 for all human and mouse cell lines in either Fig 1 or supplementary.

2. Fig 2: It would have been a better comparison to see the effect of knocking down the sST2 and ST2l individually instead of employing sh#3 shRNA which collectively knocks down both ST2l and sST2. Have authors used any shRNA to knock down ST2l individually? If so, please include the data to draw a better comparison and so the effect on tumor volume and tumor weight.

3. In Fig 3E and F the tumor volume and weight is approximately half fold in samples where sST2 is rescued as compared to control. Please discuss.

4. Fig 4A shows expression of IL-33 mRNA by RT-PCR. Since sST2 act as IL-33 sink, authors should show the expression of secretory IL-33 which is expected to up-regulate in sh#5 as compared to shCont.

Since idea of the paper is, Cancer cell-derived sST2 enhances tumor growth through upregulation of CXCL3 via inhibition of IL-33-ST2L signaling in the tumor microenvironment of pancreatic cancer, the expression of secretory IL-33 in sh#5 and shCont would be an essential piece of data to corroborate the finding in IL-33 knock out mice wherein decoy receptor seems to has no effect. Please include the said figure in Fig 4.

Reviewer #2: Here, Takenaga and colleagues conducted an in vivo study to explore the role of cancer cell derived IL-33 decoy receptor sST2 in pancreatic tumor growth. This is an interesting study as this one relatively new in pancreatic cancer and almost unexplored. The author precisely investigated the involvement of decoy receptor in tumor progression. However, in my opinion the manuscript has some shortcomings. Below I have provided numerous remarks. Please address the following concern.

Comments:

1. What is the source of IL-33 in TME. Is it pancreatic stellate cell or other cell types?

2. As a ligand, is IL-33 regulate the secretion of receptor sST2 from pancreatic cancer cell? If so, which control the secretion of receptor sST2 in global IL-33 KO mice. It is interesting to see that IL-33 signaling downregulate CXCL-3 expression, what could be the possible mechanism?

3. In figure3 panel E and F, is VC and sST2 are statistically significant?

4. It will be worthy, in future to perform some coculture experiment with PANC02 cell and CAF cell to better understand their cytokine/chemokine crosstalk in TME.

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Reviewer #1: Yes: Utpreksha Vaish

Reviewer #2: Yes: kousik kumar kesh

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Cancer cell-derived interleukin-33 decoy receptor sST2 enhances orthotopic tumor growth in a murine pancreatic cancer model

PONE-D-19-33979R1

Dear Dr. Takenaga,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

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With kind regards,

Sulagna Banerjee

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 #1: All comments have been addressed

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 #1: Yes

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

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 #1: Yes

Reviewer #2: Yes

**********

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 #1: Yes

Reviewer #2: Yes

**********

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 #1: Author' reply for Δct plots for ST2 and sST2-We have provided a plot showing the ΔCt values in the Supporting information (S1 Fig).

Reviewer's comment- This is actually S2 Fig. Check it throughput the manuscript and make correction if any required.

Reviewer #2: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

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Reviewer #1: Yes: Utpreksha Vaish

Reviewer #2: Yes: kousik kumar kesh