Development of a novel PIK3CA-mutated pancreatic tumor mouse model and evaluation of the therapeutic effects of a PI3K inhibitor

Yoshimasa Suzuki; Makoto Sugimori; Yushi Kanemaru; Sho Onodera; Hiromi Tsuchiya; Aya Ikeda; Ryosuke Ikeda; Yoshihiro Goda; Hiroaki Kaneko; Kuniyasu Irie; Soichiro Sue; Hideaki Ijichi; Shin Maeda

doi:10.1371/journal.pone.0326491

Peer Review History

Original SubmissionNovember 7, 2024
Decision Letter - Debasmita Dutta, Editor PONE-D-24-48176Generation of a novel PIK3CA-mutated pancreatic tumor mouse model and evaluation of the therapeutic effect of a PI3K inhibitorPLOS ONE Dear Dr. Maeda, 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. Please provide point by point responses to all the comments. Please provide detailed description of all methods and cite the essential and relevant references in the manuscript. Please submit your revised manuscript by Mar 14 2025 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 . 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Additional Editor Comment: Referees would like to see some revisions to your manuscript before it can be considered any further. Therefore, I invite you to respond to the comments and submit a revised manuscript. Please provide point by point responses to all comments. Please provide a detailed description of all methods and also cite the essential and relevant references in the manuscript. [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: Partly Reviewer #2: Yes Reviewer #3: Yes ******** 2. 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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 Reviewer #3: 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: This article reports pancreatic cancer development in PIK3CA mutant mouse model and its utility in evaluating PIK3CA targeted therapy. While the data is interesting, there are several points that needs attention and clarification... 1. Stock numbers for each mouse strain should be included in methods. 2. Cat# for all reagents, drugs, including antibodies should be included in methods. 3. Provide more details of in vivo study, how many mice per gender, per timepoint were evaluated for tumors. 4. include pancreas weights data for each mouse evaluated, all strains and all time points. 5. Compare the pancreas weights and lesion development between PC, PPC and KPC strains. Was there any difference between genders? 6. Was PC strain also evaluated at 50 and 100 days or just at 250 day? if yes what was the finding? if no ...why? 7. Is alpelisib able to inhibit mutant PIK3CA? discuss and provide reference. 8. Figs are not clear, need high resolution images for reviewing. 9. Why did they use subcutaneous model, instead of transgenic model itself for evaluating drug effect. Reviewer #2: Dear Authors, This is an interesting and novel research, and I appreciated the opportunity to review it. I commend your valuable effort to advance the scientific understanding of this significant health concern. Below, I have outlined my suggestions for enhancing this manuscript. 1. With regard to the precision of the manuscript, it is essential to cite relevant references for the facts and methods presented in the manuscript accurately. The first four comments address this requirement. Please cite the correct reference regarding the statement: “While PIK3CA mutations are relatively common in breast, gastric, and colorectal cancers”. It is not mentioned in the 10th reference. 2. Please cite references regarding the following sentence: Several mutation hotspots have been recognized in PIK3CA, and gain-of-function mutations such as E545K and H1047R are critical to carcinogenesis. 3. Please cite references regarding the following sentence: Thus, the indications of these PI3K/AKT inhibitors in pancreatic cancer are limited. However, for pancreatic cancer, which currently has few molecular therapy targets, the potential application of these PI3K/AKT inhibitors is needed to improve prognosis. 4. Please cite the reference for the mentioned guideline: “Reporting of In Vivo Experiments guidelines.” 5. I recommend substituting 'development' for 'generation' in the titles, as ‘development’ is more appropriate for the context. 6. Since you evaluated different effects of alpelisib, I recommend substituting 'effects' for 'effect' in the titles. 7. Please substitute the terms 'mouse' or 'mice' with 'murine' in the manuscript. This change enhances precision, as 'murine' can also refer to rats. 8. This part of the manuscript is related to the mice part of the materials and methods section: “While considering the humane endpoint, it was ensured that the tumors did not reach a size of 10 mm in any dimension. Mice were observed once every 3 days for the humane endpoints mentioned above. The duration of the experiments was about 250 days after birth. A total of 20 mice was used and euthanized. None of the mice was found dead during the experiments. Animal health and behavior were monitored every 3 days. All considerations for welfare were taken to minimize suffering and distress due to the creation of tumors. All animals were humanely euthanized via gradual CO2 exposure followed by cervical dislocation.” 9. Ethical statements should be included at the end of the Materials and Methods section after the Statistical Analysis section. 10. Please specify the sleep-wake cycle established for the mice. 11. Please define fetal bovine serum (FBS) when it is first mentioned. 12. Please provide a justification for the dosage and timing of alpelisib administration. 13. Please specify the limitations of your study before stating the conclusion. 14. Please include the supporting information files in which the data are available. Best regards. Reviewer #3: The PI3K/AKT pathway plays a crucial role in the progression of pancreatic cancer. However, there is no precedent for the successful use of PI3K inhibitors in the treatment of pancreatic cancer. Researchers have developed a new mouse pancreatic cancer model with the PIK3CA H1047R mutation. They established a pancreatic cancer cell line from PPC mice, and the PI3K p110α inhibitor alpelisib can inhibit the proliferation of PPC cells in vitro. This model is of great value for the subsequent search for therapeutic drugs for PI3K/AKT - driven pancreatic cancer. This study lacks direct evidence to verify the PIK3CA H1047R mutation. It is recommended that the authors provide information on the detection of PIK3CA mutations in cell lines using PCR. ****** 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: Yes: Sepideh Hajivalizadeh 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. 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Revision 1
18 Mar 2025 Author Response Point-by-point response to reviewers Dear reviewers, We would like to thank the editors and reviewers for reviewing our manuscript and their valuable comments. Response to Reviewer #1 >1. Stock numbers for each mouse strain should be included in methods. We included them in methods. >2. Cat# for all reagents, drugs, including antibodies should be included in methods. We included them in methods. >3. Provide more details of in vivo study, how many mice per gender, per timepoint were evaluated for tumors. We provided the details of in vivo study in supplementary information, table S1. PPC mice were evaluated on day 50 (three males), day 100 (three males), and day 150 (six males and four females). PC mice were on day150 (two males and one female) and day250 (four males and four females). KPC mice were at 18weeks of age, four males and four females. >4. include pancreas weights data for each mouse evaluated, all strains and all time points. We included pancreas weight data in table S1. >5. Compare the pancreas weights and lesion development between PC, PPC and KPC strains. Was there any difference between genders? Histological findings for each mouse are described in table S1. PC and PPC mice mainly developed tumors with cysts, but no cystic lesions were found in KPC mice. There were no gender difference in PPC mice (day150), PC mice >6. Was PC strain also evaluated at 50 and 100 days or just at 250 day? if yes what was the finding? if no ...why? PC strain was evaluated at 150 and 250 day. At 150 day, almost no tumors had developed in the pancreas. >7. Is alpelisib able to inhibit mutant PIK3CA? discuss and provide reference. According to the previous report (PMID: 24608574), Alpelisib (BYL719) also inhibits PIK3CA H1047R mutation. We inserted the following sentence in the fourth paragraph in the Background section and included the article in the references; “this inhibitor also inhibits PIK3CA E545K and H1047R mutation.” >8. Figs are not clear, need high resolution images for reviewing. We apologize for the inconvenience. Please see the attached PDF file. You can see the figures more clearly. >9. Why did they use subcutaneous model, instead of transgenic model itself for evaluating drug effect. In transgenic model, because it’s difficult to measure tumor size without dissecting the mice, the changes of tumor size over time cannot be measured. However, we can easily measure the size in subcutaneous model. So, subcutaneous model is adequate for the evaluation of the effects of alpelisib although the transgenic model more closely resembles human pancreatic tumors. Response to Reviewer #2 >1. With regard to the precision of the manuscript, it is essential to cite relevant references for the facts and methods presented in the manuscript accurately. The first four comments address this requirement. Please cite the correct reference regarding the statement: “While PIK3CA mutations are relatively common in breast, gastric, and colorectal cancers”. It is not mentioned in the 10th reference. We include the reference (PMID: 20535651). >2. Please cite references regarding the following sentence: Several mutation hotspots have been recognized in PIK3CA, and gain-of-function mutations such as E545K and H1047R are critical to carcinogenesis. We include the reference (PMID: 16322248). >3. Please cite references regarding the following sentence: Thus, the indications of these PI3K/AKT inhibitors in pancreatic cancer are limited. However, for pancreatic cancer, which currently has few molecular therapy targets, the potential application of these PI3K/AKT inhibitors is needed to improve prognosis. We include the references (PMID: 34503244, 28754816) >4. Please cite the reference for the mentioned guideline: “Reporting of In Vivo Experiments guidelines.” We include the reference (PMID: 32663219). >5. I recommend substituting 'development' for 'generation' in the titles, as ‘development’ is more appropriate for the context. We changed ‘generation’ to ‘development’ in the title. >6. Since you evaluated different effects of alpelisib, I recommend substituting 'effects' for 'effect' in the titles. We changed ‘effect’ to ‘effects’ in the title. >7. Please substitute the terms 'mouse' or 'mice' with 'murine' in the manuscript. This change enhances precision, as 'murine' can also refer to rats. We changed ‘murine’ to ‘mouse’ in the manuscripts. >8. This part of the manuscript is related to the mice part of the materials and methods section: “While considering the humane endpoint, it was ensured that the tumors did not reach a size of 10 mm in any dimension. Mice were observed once every 3 days for the humane endpoints mentioned above. The duration of the experiments was about 250 days after birth. A total of 20 mice was used and euthanized. None of the mice was found dead during the experiments. Animal health and behavior were monitored every 3 days. All considerations for welfare were taken to minimize suffering and distress due to the creation of tumors. All animals were humanely euthanized via gradual CO2 exposure followed by cervical dislocation.” We moved this part to the ‘mice’ part. >9. Ethical statements should be included at the end of the Materials and Methods section after the Statistical Analysis section. We moved ‘Ethical statements’ to the end of the Method section. >10. Please specify the sleep-wake cycle established for the mice. We included the following sentence in the mice section of Method: “The mice were housed under a 12-hour light/12-hour dark cycle, with lights on at 7:00 AM and off at 7:00 PM.” >11. Please define fetal bovine serum (FBS) when it is first mentioned. We defined fetal bovine serum(FBS) in the “cell culture”section in Method. 12. Please provide a justification for the dosage and timing of alpelisib administration. In the present study, alpelisib was administered at 40mg/kg for two weeks (5 out of 7 days). In the referenced study #28 (PMID: 30051890), it was administered at 45mg/kg for 15days. our dosage and timing of alpelisib administration is not exactly the same, but reasonable. >13. Please specify the limitations of your study before stating the conclusion. We added the following limitation before conclusion; “This study has several limitations. First, more than 90% of human pancreatic cancers are Kras-positive [13], but PPC mice do not exhibit Kras mutation. The phenotypes of mice with both Kras and PIK3CA mutations remain unclear, including their drug sensitivity and responses to PI3K and Kras inhibitors, and warrant further investigation. Second, the effect of alpelisib for PPC tumor was evaluated in a subcutaneous tumor model. Although the transgenic model more closely resembles human pancreatic tumors, it is difficult to measure tumor size over time. Therefore, our results may not reflect the effect in human pancreatic tumors due to differences in the tumor microenvironment. Finally, we have not performed in vivo experiments with the combination of alpelisib and a MEK inhibitor.” >14. Please include the supporting information files in which the data are available. We provided supporting information in which the data are available. Respose to Reviewer #3: >The PI3K/AKT pathway plays a crucial role in the progression of pancreatic cancer. However, there is no precedent for the successful use of PI3K inhibitors in the treatment of pancreatic cancer. Researchers have developed a new mouse pancreatic cancer model with the PIK3CA H1047R mutation. They established a pancreatic cancer cell line from PPC mice, and the PI3K p110α inhibitor alpelisib can inhibit the proliferation of PPC cells in vitro. This model is of great value for the subsequent search for therapeutic drugs for PI3K/AKT - driven pancreatic cancer. >This study lacks direct evidence to verify the PIK3CA H1047R mutation. It is recommended that the authors provide information on the detection of PIK3CA mutations in cell lines using PCR. We provided PCR picture in supplementary figure S1. We included the following sentence in “Effect of alpelisib on PPC and KPC cells in vitro” section in RESULT; “Recombination of the PIK3CA gene in PPC cells was confirmed by Polymerase Chain Reaction (Figure S1).” Attachments Attachment Submitted filename: Point by point Response.docx https://doi.org/10.1371/journal.pone.0326491.r002
Decision Letter - Debasmita Dutta, Editor PONE-D-24-48176R1Development of a novel PIK3CA-mutated pancreatic tumor mouse model and evaluation of the therapeutic effects of a PI3K inhibitorPLOS ONE Dear Dr. Maeda, 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. Thank you very much for implementing most of the suggestions from reviewers in R1. Please address the remaining one as well. In addition to that there are a couple of minor observations.1. Firstly please mention correct journal information in the cover/rebuttal letter.2. Secondly, in the first page of the manuscript there is a : 'These authors contributed equally to this work', but there was no respective indicator related to that. Please submit your revised manuscript by May 30 2025 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: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Debasmita Dutta, Ph.D. Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments: There are a couple of minor observations. Firstly correct journal name should be mentioned in the cover letter. Secondly, in the first page of the manuscript there is a phrase mentioned as 'These authors contributed equally to this work', but there was no respective indicator in the author list related to that. Please confirm. [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) ******** 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 ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: I Don't Know ****** 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 ****** 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 ****** 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: Dear authors, I appreciate your addressing my previous comments. Nevertheless, the manuscript could still benefit from some revision. Below, I have provided my comments regarding its improvement. 1. Compared to the referenced study (PMID: 30051890), the administration of alpelisib was approximately 69% lower. Please provide a rationale for this reduction. Best regards. ****** 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: Yes: Sepideh Hajivalizadeh ******** [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.0326491.r003
Revision 2
19 Apr 2025 Author Response Point-by-point response to editors and reviewers Dear editors and reviewers, We would like to thank the editors and reviewers for reviewing our manuscript and their valuable comments. Response to editors >1. Firstly, please mention correct journal information in the cover/rebuttal letter. We sincerely apologize for the error in the journal name mentioned in our cover letter. We have corrected it and submitted the revised version accordingly. >2. Secondly, in the first page of the manuscript there is a : 'These authors contributed equally to this work', but there was no respective indicator related to that. We have now added the appropriate asterisks and daggers to the author names on the title page to indicate equal contributions, in accordance with the journal's guidelines. Response to Reviewer #2 >1. Compared to the referenced study (PMID: 30051890), the administration of alpelisib was approximately 69% lower. Please provide a rationale for this reduction. Thank you for your valuable comment. The dose of alpelisib used in preclinical studies varies across reports. A previous study (PMID: 32899250) demonstrated that a dose of 15 mg/kg administered over four weeks was sufficient to achieve therapeutic efficacy. The total drug exposure in that study is comparable to that in our current study, supporting the validity of the dosing regimen we employed. Therefore, we believe our selected dose is appropriate and justified. We have cited this reference in the Methods section under the description of the subcutaneous tumor allograft model. Attachments Attachment Submitted filename: Point by point Response2.docx https://doi.org/10.1371/journal.pone.0326491.r004
Decision Letter - Debasmita Dutta, Editor PONE-D-24-48176R2Development of a novel PIK3CA-mutated pancreatic tumor mouse model and evaluation of the therapeutic effects of a PI3K inhibitorPLOS ONE Dear Dr. Maeda, 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. Thank you very much for submitting a revised version. Please follow PLOS guidelines properly to indicate corresponding author and multiple groups of equally contributed authors. Please submit your revised manuscript by Jun 07 2025 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: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Debasmita Dutta, Ph.D. Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] [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.0326491.r005
Revision 3
20 May 2025 Author Response Point-by-point response to editors Dear editors, We would like to thank the editors for reviewing our manuscript and their valuable comments. Response to editors >Please follow PLOS guidelines properly to indicate corresponding author and multiple groups of equally contributed authors. We sincerely apologize for the incorrect statement regarding the contributing authors. The author contributions have been accurately entered in the submission form, and we have now removed the equal contribution note from the manuscript. Attachments Attachment Submitted filename: Point by point Response 3.docx https://doi.org/10.1371/journal.pone.0326491.r006
Decision Letter - Debasmita Dutta, Editor Development of a novel PIK3CA-mutated pancreatic tumor mouse model and evaluation of the therapeutic effects of a PI3K inhibitor PONE-D-24-48176R3 Dear Dr. Maeda, 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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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, Debasmita Dutta, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Thank you for addressing review comments. Reviewers' comments: https://doi.org/10.1371/journal.pone.0326491.r007
Formally Accepted
Acceptance Letter - Debasmita Dutta, Editor PONE-D-24-48176R3 PLOS ONE Dear Dr. Maeda, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, 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 customercare@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. Debasmita Dutta Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0326491.r008

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