Tumor microenvironment predicts local tumor extensiveness in PD-L1 positive nasopharyngeal cancer

Soehartati A. Gondhowiardjo; Handoko; Marlinda Adham; Lisnawati Rachmadi; Henry Kodrat; Demak Lumban Tobing; I. Made Haryoga; Agustinus Gatot Dwiyono; Yoseph Adi Kristian; Tiara Bunga Mayang Permata

doi:10.1371/journal.pone.0230449

Peer Review History

Original SubmissionDecember 3, 2019
7 Feb 2020 Decision Letter - Salvatore V Pizzo, Editor PONE-D-19-33450 Tumor Microenvironment Predicts Local Tumor Extensiveness in PD-L1 Positive Nasopharyngeal Cancer PLOS ONE Dear Mr. Handoko, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. As you can see, both reviewers have made suggestions for minor changes to the manuscript. We would request your consideration of these changes. We would appreciate receiving your revised manuscript by Mar 23 2020 11:59PM. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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Reviewer #1: Yes Reviewer #2: Yes ****** 3. 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 ****** 4. 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 ****** 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: In this manuscript, the authors found that the PD-L1 was highly expressed in 25 NPC samples. They also revealed that denser local tumor infiltrating immune cells population have relatively much small local tumor volume. Thus, tumor infiltrating immune cells play an important role in tumor progression. The detection of tumor infiltrating immune cells may offer valuable prognostic information for NPC. This manuscript is well-written and the figures are well-presented. I only have one comment. I wonder if these NPC patents are receiving chemotherapy during this project. Chemotherapy or other therapies may have an impact on the distribution and density of tumor infiltrating immune cells. The authors should clarify this in the manuscript. Reviewer #2: 1.Local tumor volume can be measured by CT or MRI scan exactly. There is no need to "predict" it. In this research the authors indicated the relationship between tumor volume and its microenviroment, changing the title into, for example, "Tumor microenvironment charactaristics in different tumor extensiveness in PD-L1 positive nasopharyngeal cancer" would more appropriate. 2.Did the enrolled patients received any treatment before biopsy? Medical treatment can influence the tumor progression and its microenviroment and further affect the final results. 3.In line 153 Did the enrolled patients have any hematological disease or infectious complications when taking peripheral blood count? These factors can affect the peripheral blood components and authors should illuminate this. 4.In line 174 It may be insufficient to use HE dye to evaluate tumor infiltrating immune cells level because it is too subjective. CD45 is widely expressed in all kinds of immune cells, so I suggest that CD45 staining is need to help evaluating the infiltrating level. 5.In line 187 "imaging selected" means "selected imaging" ? 6.PD-L1 is an important immunosupression molecular. So what is the relationship between PD-L1 expression and tumor infiltrating immune cells? In other kinds of cancers, PD-L1 often has higher expression in "hot tumor". ****** 6. 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Revision 1
14 Feb 2020 Author Response Reviewer #1: In this manuscript, the authors found that the PD-L1 was highly expressed in 25 NPC samples. They also revealed that denser local tumor infiltrating immune cells population have relatively much small local tumor volume. Thus, tumor infiltrating immune cells play an important role in tumor progression. The detection of tumor infiltrating immune cells may offer valuable prognostic information for NPC. This manuscript is well-written and the figures are well-presented. I only have one comment. I wonder if these NPC patents are receiving chemotherapy during this project. Chemotherapy or other therapies may have an impact on the distribution and density of tumor infiltrating immune cells. The authors should clarify this in the manuscript. Thank you for the comment. The answer was no. All subjects included in this study did not have any prior cancer treatment. It’s true that some systemic treatments might impact tumor microenvironment. Thank you for pointing it out for us, now in our revised manuscript we have added a statement clarifying that issue. Reviewer #2: 1.Local tumor volume can be measured by CT or MRI scan exactly. There is no need to "predict" it. In this research the authors indicated the relationship between tumor volume and its microenviroment, changing the title into, for example, "Tumor microenvironment charactaristics in different tumor extensiveness in PD-L1 positive nasopharyngeal cancer" would more appropriate. Thank you for the comment and suggestion. Yes it is true that we can obtain exact tumor volume by CR or MRI scan. Your suggestion “Tumor microenvironment characteristics in different tumor extensiveness in PD-L1 positive nasopharyngeal cancer" was great, however, the term “in different tumor extensiveness” could be connoted by some people as different disease stages. While actually in our study we used exact tumor volume. We used “predict” because we wanted to give the readers an upfront impression that tumor microenvironment has prognostic role when just glancing at our title. We refereed “predict” here as predicting role of tumor microenvironment. Therefore, we actually tend to stick to our original title. But we are still open to discussion if you have any other comments or ideas. 2.Did the enrolled patients received any treatment before biopsy? Medical treatment can influence the tumor progression and its microenviroment and further affect the final results. No, all patients did not receive any prior treatment. We have revised and included a clear statement in our revised manuscript that all subjects enrolled in our study did not ever received any kind of treatment before. 3.In line 153 Did the enrolled patients have any hematological disease or infectious complications when taking peripheral blood count? These factors can affect the peripheral blood components and authors should illuminate this. No, all patients enrolled did not have any hematological disease nor infections. The routine peripheral blood checks was part of our institution protocol. All patients would have the blood checked before biopsy and every week during treatment. Furthermore, the patient would not be biopsied if they had any kind of infection. We have also included a statement in our revised manuscript stating that all patients enrolled did not have any other severe diseases such hematological diseases or infectious diseases. 4.In line 174 It may be insufficient to use HE dye to evaluate tumor infiltrating immune cells level because it is too subjective. CD45 is widely expressed in all kinds of immune cells, so I suggest that CD45 staining is need to help evaluating the infiltrating level. Yes, we were aware about the subjectivity. Therefore, we have actually made a precautionary step by having 2 independent pathologists blindly reviewing the HE slides. Nevertheless, we are very happy with your suggestion. Next time we will add CD45 staining to help us tackle this subjectivity issue. 5.In line 187 "imaging selected" means "selected imaging" ? Yes, we meant selected imaging. We have revised it in our revised manuscript. Thanks for point it out for us. 6.PD-L1 is an important immunosupression molecular. So what is the relationship between PD-L1 expression and tumor infiltrating immune cells? In other kinds of cancers, PD-L1 often has higher expression in "hot tumor". There was no relationship found between PD-L1 expression and tumor infiltrating immune cells in our study. We were aware that some tumor did have relationship exist between those two. We have added a statement in our revised manuscript describing the absence of relationship between PD-L1 and tumor infiltrating immune cells in our NPC samples. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0230449.r002
2 Mar 2020 Decision Letter - Salvatore V Pizzo, Editor Tumor Microenvironment Predicts Local Tumor Extensiveness in PD-L1 Positive Nasopharyngeal Cancer PONE-D-19-33450R1 Dear Dr. Handoko, 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. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. With kind regards, Salvatore V Pizzo 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 ******** 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: The authors have well addressed all my comments in the revised version of the manuscript. I have no further comment. Reviewer #2: (No Response) ****** 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. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No https://doi.org/10.1371/journal.pone.0230449.r003
Formally Accepted
6 Mar 2020 Acceptance Letter - Salvatore V Pizzo, Editor PONE-D-19-33450R1 Tumor Microenvironment Predicts Local Tumor Extensiveness in PD-L1 Positive Nasopharyngeal Cancer Dear Dr. No Last Name: I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Salvatore V Pizzo Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0230449.r004

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