PI3k inhibitors (BKM120 and BYL719) as radiosensitizers for head and neck squamous cell carcinoma during radiotherapy

Fu-Cheng Chuang; Chih-Chun Wang; Jian-Han Chen; Tzer-Zen Hwang; Shyh-An Yeh; Yu-Chieh Su

doi:10.1371/journal.pone.0245715

Peer Review History

Original SubmissionJuly 6, 2020
14 Aug 2020 Decision Letter - Salvatore V Pizzo, Editor PONE-D-20-20798 PI3k inhibitors (BKM120 and BYL719) as radiosensitizers for head and neck squamous cell carcinoma during radiotherapy PLOS ONE Dear Dr. Su, 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. Serious issues have been raised with respect to your manuscript. Indeed, one reviewer suggested rejection. Nevertheless, we would be willing to consider a revised manuscript if the authors feel they can address the criticisms. Please ensure that your decision is justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact. ============================== Please submit your revised manuscript by Sep 28 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Salvatore V Pizzo Academic Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Please provide additional information about each of the cell lines used in this work, including any quality control testing procedures (authentication, characterisation, and mycoplasma testing). For more information, please see http://journals.plos.org/plosone/s/submission-guidelines#loc-cell-lines 3. Thank you for stating the following financial disclosure: "NO - Include this sentence at the end of your statement: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." At this time, please address the following queries: a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution. b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” c) If any authors received a salary from any of your funders, please state which authors and which funders. d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.” Please include your amended statements within your cover letter; we will change the online submission form on your behalf [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. 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: No Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? 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: No 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: The authors present an interesting analysis of the use of PI3K inhibitors to sensitize OSCC models to radiation therapy that has some novelty in this field. At this point, the manuscript lacks substantial supporting experiments to warrant publication, and some recommendations to help the authors improve the manuscript are listed below. 1) Inclusion of pharmacodynamic analysis of PI3K effector signaling in the models presented, with the doses used (which are often very high). 2) Inclusion of pharmacodynamic analysis of DNA damage responses (e.g. gH2AX, COMET assays) following radiation, as well as more detailed analysis of mechanisms of cell death. 3) In vivo modeling of the various combinations. 4) Supporting genetic approaches to model changes to PI3K isoforms and their regulation of the DNA damage response. 5) Evaluation of alternative modes of DNA damage to determine if the phenotype is unique to radiation and/or DNA double strand break induction. Reviewer #2: This study describes the effects of the PI3K� inhibitors Alpelisib (BYL719) and Buparlisib (BKM120) on the radiation sensitivity of head and neck cancer cells. Although these studies are of some interest, they are limited in scope and descriptive. There are no mechanistic studies. In addition, the lack of in vivo studies makes it difficult to know if these agents and the combination with the mTOR inhibitor will translate into an actual treatment in patients. 1) Do the authors know if the concentrations used in the in vitro experiments are clinically relevant? What concentrations are found in patients receiving these drugs? 2) The use of a single radiation dose (Figures 2 and 3) give limited information on overall radiation sensitivity. This is usually best assessed by performing a full cell survival curve of control and 3-4 radiation doses. 3) Did the agents actually inhibit PI3K� under the conditions that were investigated for radiosensitization? This would be the best way to verify that the effects. 4) Why did the authors choose combine these inhibitors with an mTOR inhibitor. 5) The MTT like assay is useful for screening large number of conditions rapidly but is not as accurate as a clonogenic assay for detailed assessment of radiosensitization 6) The clinical applicability of these combinations is difficult to assess with in vitro experiments alone. ****** 6. 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 [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0245715.r001
Revision 1
2 Oct 2020 Author Response 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 present an interesting analysis of the use of PI3K inhibitors to sensitize OSCC models to radiation therapy that has some novelty in this field. At this point, the manuscript lacks substantial supporting experiments to warrant publication, and some recommendations to help the authors improve the manuscript are listed below. 1) Inclusion of pharmacodynamic analysis of PI3K effector signaling in the models presented, with the doses used (which are often very high). Our response: As suggested by reviewer, we have considered the in vitro time-course pharmacodynamic analysis of the PI3K inhibitor, but we decided the assay will be performed in vivo study which may provide more information for the clinical trials. However, we still work on the development of the animal model because of the radiotherapy assay being not easy to perform. 2) Inclusion of pharmacodynamic analysis of DNA damage responses (e.g. gH2AX, COMET assays) following radiation, as well as more detailed analysis of mechanisms of cell death. Our response: Previous study has demonstrated that BYL719 combined with RT induced the tumor γH2AX nuclear formation in HNSCC animal model (1). BKM120 also showed enhanced DNA damage activity in hepatocellular carcinoma cells (2). The detailed mechanisms we will perform in the HNSCC animal model in the further study. Reference 1. Mizrachi A, Shamay Y, Shah J, Brook S, Soong J, Rajasekhar VK, et al. Tumour-specific PI3K inhibition via nanoparticle-targeted delivery in head and neck squamous cell carcinoma. Nature communications. 2017;8:14292. 2. Liu WL, Gao M, Tzen KY, Tsai CL, Hsu FM, Cheng AL, et al. Targeting Phosphatidylinositide3-Kinase/Akt pathway by BKM120 for radiosensitization in hepatocellular carcinoma. Oncotarget. 2014;5(11):3662-72. 3) In vivo modeling of the various combinations. Our response: As suggested by reviewer, we understand the importance of animal model for the assessing the clinical applicability. The animal model commonly used the immunodeficiency animal model to assess the activity of the inhibitors, but the model is under the lack of host immunity-tumor cell interaction that may not truly reflect the activity of inhibitors in the in vivo assay. Furthermore, the application of radiotherapy in animal model is not easy to perform. Therefore, we are searching the best solution for the animal model in the further study. 4) Supporting genetic approaches to model changes to PI3K isoforms and their regulation of the DNA damage response. Our response: As suggested by reviewer, the expression of γH2AX and PARP, which represent the DNA damage response, by western blotting will be checked in the combination treatment of PI3K inhibitor and radiotherapy in HNSCC animal model in the further study. 5) Evaluation of alternative modes of DNA damage to determine if the phenotype is unique to radiation and/or DNA double strand break induction. Our response: The alternative modes of DNA damage will be identified by performing NGS assay to comprehensively investigate the mechanism in the combination treatment of PI3K inhibitor and radiotherapy in HNSCC animal model in the further study. Reviewer #2: This study describes the effects of the PI3K� inhibitors Alpelisib (BYL719) and Buparlisib (BKM120) on the radiation sensitivity of head and neck cancer cells. Although these studies are of some interest, they are limited in scope and descriptive. There are no mechanistic studies. In addition, the lack of in vivo studies makes it difficult to know if these agents and the combination with the mTOR inhibitor will translate into an actual treatment in patients. 1) Do the authors know if the concentrations used in the in vitro experiments are clinically relevant? What concentrations are found in patients receiving these drugs? Our response: Yes, we know the relevant between the in vitro experiments and clinical usage dose. The concentration of alpelisib used in patients is at a dose of 300 mg per day, and the concentration of buparlisib used in patients is 100 mg per day. 2) The use of a single radiation dose (Figures 2 and 3) give limited information on overall radiation sensitivity. This is usually best assessed by performing a full cell survival curve of control and 3-4 radiation doses. Our response: The suggestion by reviewers was ideal to support more information. Our unshown results demonstrated that different radiation doses dose-dependent reduced the cell growth when treated alone or combination treatment with PI3K inhibitors. We added the result into the manuscript as figure 3. 3) Did the agents actually inhibit PI3K� under the conditions that were investigated for radiosensitization? This would be the best way to verify that the effects. Our response: The suggestion by reviewers was ideal to confirm the effect of PI3K inhibitor. The pan-PI3K inhibitor buparlisib (BKM120) and the PI3K inhibitor alpelisib (BYL719) are developed by Novartis Oncology for clinical patients, and the specific of the inhibitors against PI3K is definite under the conditions. 4) Why did the authors choose combine these inhibitors with an mTOR inhibitor. Our response: The PI3K/AKT/mTOR signaling pathway is a common mechanism to initiate the survival and proliferation. Therefore, the combination treatment of PI3K and mTOR inhibitor may boost the anticancer effect in the OSCC cells. 5) The MTT like assay is useful for screening large number of conditions rapidly but is not as accurate as a clonogenic assay for detailed assessment of radiosensitization Our response: As suggested by reviewer, the MTT assay in present study was the primary experiments to determine the available concentrations of the inhibitors for the further assay. Using clonogenic assay to investigate the appropriate concentrations is more time consuming. 6) The clinical applicability of these combinations is difficult to assess with in vitro experiments alone. Our response: As suggested by reviewer, we understand the importance of animal model for the assessing the clinical applicability. The animal model commonly used the immunodeficiency animal model to assess the activity of the inhibitors, but the model is under the lack of host immunity-tumor cell interaction that may not truly reflect the activity of inhibitors in the in vivo assay. Furthermore, the application of radiotherapy in animal model is not easy to perform. Therefore, we are searching the best solution for the animal model in the further study. Attachments Attachment Submitted filename: response to reviewers_BKM_plos one.docx https://doi.org/10.1371/journal.pone.0245715.r002
16 Nov 2020 Decision Letter - Salvatore V Pizzo, Editor PONE-D-20-20798R1 PI3k inhibitors (BKM120 and BYL719) as radiosensitizers for head and neck squamous cell carcinoma during radiotherapy PLOS ONE Dear Dr. Su, 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. One reviewer is concerned that mechanistic studies are lacking and the study is purely descriptive. Indeed this reviewer recommended rejection. Nevertheless, we would consider a revised manuscript but these issues should be addressed. Please submit your revised manuscript by Dec 31 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Salvatore V Pizzo Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] 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 #2: (No Response) Reviewer #3: 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 #2: Partly Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: The authors have responded to some comments but not some critical ones. Comment 1) - the question was about concentrations and the authors have answered in mg Comments 5 and 6). this reviewer agrees that performing clonogenic assays and animal experiments is time consuming. But they provide important information concerning the clinical applicability of this combination. Reviewer #3: This study aimed to identify PI3K inhibitors that can enhance radiosensitivity in head and neck cancer cells. The authors showed that a combination of BKM120 or BYL719 with irradiation leads to an increase in radiosensitivity in HNSCC cells. After a meticulous review of the manuscript, I found that the current study could be pertinent and informative for the readers this journal. However, I think that there is some room for improvements before a further consideration for publication. At this point there is a need for minor revision before the manuscript could be reconsidered for publication in the journal. According to my assessments, there are some points to clarify and revise: In addition to radiotherapy, surgery is a widely used therapeutic option in patients with HNSCC. This should not remain unmentioned in the discussion, since surgery is superior to radiation therapy in early stages. Furthermore, in advanced tumors, radiotherapy is used in combination with chemotherapy and not as the sole modality. The authors should take a more differentiated view on the therapy of HNSCC and side effects of radiotherapy in the discussion section. I highly recommend investigating the effects of BKM120 and BYL719 in vivo in the near future. ****** 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 #2: No Reviewer #3: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0245715.r003
Revision 2
23 Dec 2020 Author Response 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 #2: The authors have responded to some comments but not some critical ones. Comment 1) - the question was about concentrations and the authors have answered in mg Our response: As suggested by reviewer, the concentrations of the inhibitors in patients is associated with how many mg of the inhibitor that patient took, and the inhibitors concentration in the patient plasma appear dynamic which was showed as pharmacokinetics. For example, the alpelisib used in patients at a dose of 350 mg per day in cycle 1 showed the median maximum plasma concentration (Cmax) is 3490 ng/ml. The buparlisib used in patients at a dose of 100 mg per day in cycle 1 showed the Cmax is 1080 ng/ml. Comments 5 and 6). this reviewer agrees that performing clonogenic assays and animal experiments is time consuming. But they provide important information concerning the clinical applicability of this combination. Our response: As suggested by reviewer, we understand and agree that the animal model is important information for clinical applicability of the combination. However, previous studies had indicated that the PI3K/Akt/mTOR pathway inhibitors plus radiotherapy were ideal strategy for lung cancer and breast cancer (1, 2). Furthermore, several PI3K/Akt/mTOR inhibitors including alpelisib and buparlisib has been involved in clinical trials, indicating that the use of the inhibitor is available (3, 4). The combination treatment of PI3K/Akt/mTOR inhibitor and radiotherapy has also been identify that the strategy is clinical applicable(1). On the other hand, the use of the hospital facility was limited because of the novel coronavirus disease, which has influenced the progress of the study. In the near future, we will proceed the in vivo analysis to provide the direct evidence. Reference: 1. Chen K, Shang Z, Dai AL, Dai PL. Novel PI3K/Akt/mTOR pathway inhibitors plus radiotherapy: Strategy for non-small cell lung cancer with mutant RAS gene. Life sciences. 2020;255:117816. 2. DuRoss AN, Neufeld MJ, Landry MR, Rosch JG, Eaton CT, Sahay G, et al. Micellar Formulation of Talazoparib and Buparlisib for Enhanced DNA Damage in Breast Cancer Chemoradiotherapy. ACS applied materials & interfaces. 2019;11(13):12342-56. 3. Rodon J, Brana I, Siu LL, De Jonge MJ, Homji N, Mills D, et al. Phase I dose-escalation and -expansion study of buparlisib (BKM120), an oral pan-Class I PI3K inhibitor, in patients with advanced solid tumors. Investigational new drugs. 2014;32(4):670-81. 4. Beck JT, Ismail A, Tolomeo C. Targeting the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway: an emerging treatment strategy for squamous cell lung carcinoma. Cancer treatment reviews. 2014;40(8):980-9. Reviewer #3: This study aimed to identify PI3K inhibitors that can enhance radiosensitivity in head and neck cancer cells. The authors showed that a combination of BKM120 or BYL719 with irradiation leads to an increase in radiosensitivity in HNSCC cells. After a meticulous review of the manuscript, I found that the current study could be pertinent and informative for the readers this journal. However, I think that there is some room for improvements before a further consideration for publication. At this point there is a need for minor revision before the manuscript could be reconsidered for publication in the journal. According to my assessments, there are some points to clarify and revise: In addition to radiotherapy, surgery is a widely used therapeutic option in patients with HNSCC. This should not remain unmentioned in the discussion, since surgery is superior to radiation therapy in early stages. Furthermore, in advanced tumors, radiotherapy is used in combination with hemotherapy and not as the sole modality. Theauthors should take a more differentiated view on the therapy of HNSCC and side effects of radiotherapy in the discussion section. I highly recommend investigating the effects of BKM120 and BYL719 in vivo in the near future. Our response: As suggested by reviewer, we added more discussion about the importance of surgery and the side effect when patients treated with radiotherapy or chemotherapy in the discussion section. We will proceed the in vivo analysis soon. Because of the novel coronavirus disease, the use of the hospital facility was limited that also influence the progress of the study. Attachments Attachment Submitted filename: 20201207_Response to reviewer.docx https://doi.org/10.1371/journal.pone.0245715.r004
7 Jan 2021 Decision Letter - Salvatore V Pizzo, Editor PI3k inhibitors (BKM120 and BYL719) as radiosensitizers for head and neck squamous cell carcinoma during radiotherapy PONE-D-20-20798R2 Dear Dr. Su, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. 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 help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- 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. 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 #2: (No Response) Reviewer #3: 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 #2: Yes Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: (No Response) Reviewer #3: (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 #2: No Reviewer #3: No https://doi.org/10.1371/journal.pone.0245715.r005
Formally Accepted
11 Jan 2021 Acceptance Letter - Salvatore V Pizzo, Editor PONE-D-20-20798R2 PI3k inhibitors (BKM120 and BYL719) as radiosensitizers for head and neck squamous cell carcinoma during radiotherapy Dear Dr. Su: I'm 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 let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, 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. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. 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.0245715.r006

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