Insulin-like growth factor 1 receptor expression correlates with programmed death ligand 1 expression and poor survival in non-small cell lung cancer

Hiroaki Nagamine; Masakazu Yashiro; Megumi Mizutani; Akira Sugimoto; Yoshiya Matsumoto; Yoko Tani; Kenji Sawa; Hiroyasu Kaneda; Kazuhiro Yamada; Tetsuya Watanabe; Kazuhisa Asai; Satoshi Suzuki; Tomoya Kawaguchi

doi:10.1371/journal.pone.0297397

Peer Review History

Original SubmissionJanuary 4, 2024
2 Jul 2024 Decision Letter - Huei-Wen Chen, Editor PONE-D-23-43471Insulin-like Growth Factor 1 Receptor Expression Correlates with Programmed Death Ligand 1 Expression and Poor Survival in Non-small Cell Lung CancerPLOS ONE Dear Dr. Yashiro, 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 submit your revised manuscript by Aug 16 2024 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, Huei-Wen Chen, Ph.D. 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. In the online submission form, you indicated that "The data underlying the results presented in the study are available from correspondence author "Masakazu Yashiro"." All PLOS journals now require all data underlying the findings described in their manuscript to be freely available to other researchers, either 1. In a public repository, 2. Within the manuscript itself, or 3. Uploaded as supplementary information. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If your data cannot be made publicly available for ethical or legal reasons (e.g., public availability would compromise patient privacy), please explain your reasons on resubmission and your exemption request will be escalated for approval. 3. 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: Dear Prof. Yashiro, Many thanks for your patience. Your manuscript entitled “ Insulin-like Growth Factor 1 Receptor Expression Correlates with Programmed Death Ligand 1 Expression and Poor Survival in Non-small Cell Lung Cancer” has now been seen by 2 referees, whose comments are enclosed as below. In light of these comments, we would be interested in considering a minor revised version that addresses the comments of the referees. The study presents compelling evidence that IGF1R expression is associated with poorer survival outcomes and higher PD-L1 expression in NSCLC patients, particularly in early stages. This highlights the potential of IGF1R as a prognostic marker and its possible role in shaping perioperative treatment strategies. Further research is needed to validate these findings, including IHC staining of the co-positive of IGF1R and PDL1? Immune cells infiltration? explore underlying mechanisms; especially, this study suggests a link between IGF1R and worse outcomes but does not delve into the underlying mechanisms. Further discussion should be added and aim to elucidate how IGF1R contributes to tumor progression, resistance to therapies, cancer stem cells, and might contribute on onco-immune regulations. and assess the value of this in early stage of targeting IGF1R in NSCLC. [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: Yes Reviewer #2: Yes ******** 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: 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: The present manuscript “Insulin-like Growth Factor 1 Receptor Expression Correlates with Programmed Death Ligand 1 Expression and Poor Survival in Non-small Cell Lung Cancer“ by Nagamine et al. describes the results of a large-scale, long-term observational study showing that IGF1R expression is associated with PD-L1 expression in the tumor and translates into worse postoperative recurrence and long-term prognosis in lung cancer. The manuscript is well written and the methods are sound. Combined ICI and IGF1R inhibition as possible therapeutic intervention (as mentioned by the authors) should be discussed in further detail based on current literature. The underlying mechanistic circuitries should be also discussed in further detail. Future correlative studies should focus on IGF1R expression and responsiveness to immune checkpoint blockade. Reviewer #2: 1. In Keyword section IGF1R, NSCLC, OS, PD-L1, RFS should be written in full along with abbreviations for easy database search. 2. Few grammatical mistakes are there in manuscript. Please check throughout the manuscript ****** 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.0297397.r001
Revision 1
11 Jul 2024 Author Response Editorial Office PLOS ONE Dear Editor-in-chief Reference number: PONE-D-23-43471 Title: Insulin-like Growth Factor 1 Receptor Expression Correlates with Programmed Death Ligand 1 Expression and Poor Survival in Non-small Cell Lung Cancer Authors：Hiroaki Nagamine, et al. We thank you and the reviewers for your thoughtful suggestions and insights. The manuscript has benefited from these insightful suggestions. I look forward to working with you and the reviewers to move this manuscript closer to publication in the PLOS ONE. The manuscript has been rechecked and the necessary changes have been made in accordance with the reviewers’ suggestions. The responses to all comments have been prepared and attached herewith. We have added a discussion based on the editor and reviewers’ comments. As you pointed out, clarifying whether IGF1R expression is related to immune cell infiltration is an important future task, and we have added this point to the limitations section. In addition, as it would be beneficial, we have also added HE-stained images to Figure 1. In addition, some RFS were mistakenly listed as PFS, which has been corrected (p14, line 174 and 176). The changes are highlighted in yellow. All authors are aware of the content of this manuscript. Thank you for your consideration. I look forward to hearing from you. Sincerely, Masakazu Yashiro Molecular Oncology and Therapeutics, Osaka Metropolitan University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka 545-8585, Japan Email: i21496f@omu.ac.jp Reply to Reviewer 1 � Combined ICI and IGF1R inhibition as possible therapeutic intervention (as mentioned by the authors) should be discussed in further detail based on current literature. ⇒ [Response] Thank you for pointing out this critical point. Although clinical trials have not yet been conducted, several studies in mouse models have reported that combining an IGF1R inhibitor and immune checkpoint inhibitors (ICIs), such as PD-1 inhibitors, synergistically enhances antitumor effects. IGF1R influences the tumor microenvironment in various ways, and the inhibition of IGF1R is known to affect the activation of cytotoxic CD8+ T cells, which are important for the efficacy of ICIs. Therefore, combining IGF1R inhibitors with ICIs may enhance antitumor effects. This notion has been supported by recent studies, which we have referenced accordingly. ・However, recent mouse experiments have revealed that reduced IGF1R expression enhances the effects of ICI [23], and that IGF1R expression correlates with the tumor microenvironment [24], including immune cells. Combining IGF1R inhibitors with ICI may hold promise for future therapies, especially as perioperative treatment for early-stage lung cancer. ↓ ・However, recent experiments in mice have demonstrated that the combination of an IGF1R inhibitor and an anti-PD-1 antibody synergistically inhibits tumor growth [32, 33]. IGF1R expression is correlated with the tumor microenvironment and inhibits the activation of effector cytotoxic CD8+ T cells [34, 35]. Therefore, inhibition of the IGF1R pathway has the potential to enhance the effectiveness of ICI by activating effector cytotoxic CD8 T cells [36]. From those results, Combining IGF1R inhibitors with ICI may hold promise for future therapies, especially as perioperative treatment for early-stage lung cancer. � The underlying mechanistic circuitries should be also discussed in further detail. ⇒ [Response] Thank you for pointing out the important details. We have added discussions on the underlying mechanisms, citing various studies. Specifically, we have added discussions on how IGF1R is involved in cell proliferation, the high recurrence rates in early-stage lung cancer, and how IGF1R expression is related to chemoresistance. ・The IGF1R-positive group also showed significantly larger tumor sizes. IGF1R is a factor associated with tumor growth [4], consistent with a relationship between IGF1R expression and tumor size. In addition, IGF1R expression was associated with shorter RFS and OS, particularly in early-stage lung cancer. ↓ ・The IGF1R-positive group also showed significantly larger tumor sizes. IGF1R is a factor associated with tumor growth [4]. Upon binding with its ligands IGF1 or IGF2, IGF1R activates downstream signaling pathways such as the phosphoinositide 3-kinase (PI3K)/Akt and extracellular signal-regulated kinase (ERK) pathways, which leads to tumor cell proliferation [18-20]. This is consistent with the result of the relationship between IGF1R expression and tumor size. In addition, IGF1R expression was associated with shorter RFS and OS, particularly in early-stage lung cancer. It has been known that stage 1 has a lower frequency and amount of postoperative molecular residual disease (MRD) compared to stage 2 or 3 in lung cancer [21]. Although the immune system may eliminate minimal MRD in early-stage lung cancer, our results suggested that IGF1R expression was associated with PD-L1 expression, which might have allowed cancer cells to escape immune surveillance [22, 23] and increased the likelihood of relapse in IGF1R-positive patients. Additionally, IGF1R expression may facilitate tumor growth, leading to earlier recurrence compared to IGF1R-negative cases. Also, IGF1R expression has been known to correlate with the expression of ATP–binding cassette subfamily G member 2 (ABCG2) [24], a potential marker for cancer stem cells [25] and involved in chemoresistance [26]. These findings might explain the treatment resistance of recurrent tumors and may account for the worse OS observed in early-stage lung cancer with IGF1R positivity, where recurrence is more frequent. � Future correlative studies should focus on IGF1R expression and responsiveness to immune checkpoint blockade. ⇒ [Response] Thank you for pointing out these important details. There are currently no studies evaluating the relationship between IGF1R expression and the effects of ICI. This is a significant issue for the future. Therefore, based on your suggestions, we have added the following text as a limitation. ・Furthermore, prospective clinical trials on the combination therapy of IGF1R inhibitors and ICIs are warranted. The relationship between IGF1R expression and the effects of ICI was previously unknown and requires further study. In addition, future research should investigate whether IGF1R expression is related to immune cell infiltration and elucidate the underlying mechanisms. Reply to Reviewer 2 1. In Keyword section IGF1R, NSCLC, OS, PD-L1, RFS should be written in full along with abbreviations for easy database search. ⇒ [Response] Thank you for your comment. I have added the full notation of each abbreviation in the keywords section. ・Keywords: IGF1R, lung cancer, NSCLC, OS, PD-L1, RFS ↓ ・Keywords: IGF1R, insulin-like growth factor 1 receptor, lung cancer, NSCLC, non-small cell lung cancer, OS, overall survival, PD-L1, programmed cell death ligand 1, RFS, recurrence free survival 2. Few grammatical mistakes are there in manuscript. Please check throughout the manuscript. ⇒ [Response] Thank you for pointing this out. We have reviewed the text again and made some changes.　Specifically, they are as follows. p11, line 63 ・Moreover, smoking is known to be involved in tumor-induced IGF1R expression [10]. ↓ ・Moreover, smoking has been known to be involved in tumor-induced IGF1R expression [10]. p13, line 87 ・This study was approved by the Osaka City university Ethics Committee (reference number 2019-006). ↓ ・This study was approved by the Osaka City University Ethics Committee (reference number 2019-006). p13, line 89 ・This study has been conducted according to the principles of the declaration of Helsinki. ↓ ・This study was conducted according to the principles of the Declaration of Helsinki. p15, line 111 to 115 ・Multiple logistic regression analysis was utilized to analyze the relationship between IGF1R and PD-L1 expression, incorporating factors age, sex, smoking history, Eastern Cooperative Oncology Group Performance Status (ECOG PS), histology, pStage, pleural invasion, lymphatic invasion, and vascular invasion. ↓ Multiple logistic regression analysis was utilized to analyze the relationship between IGF1R and PD-L1 expression, incorporating age, sex, smoking history, Eastern Cooperative Oncology Group Performance Status (ECOG PS), histology, pStage, pleural invasion, lymphatic invasion, and vascular invasion as factors. p15, line 115 to 117 ・The Mann–Whitney U test was applied to evaluate differences in smoking index or maximum tumor diameter between the IGF1R-positive and negative groups. ↓ ・The Mann–Whitney U test evaluated differences in smoking index or maximum tumor diameter between the IGF1R-positive and negative groups. p25, line 218 to 219 ・However, due to the favorable prognosis of early-stage lung cancer, it was less amenable to the benefits of neoadjuvant and adjuvant chemotherapy. ↓ ・However, due to the favorable prognosis of early-stage lung cancer, it is less amenable to the benefits of neoadjuvant and adjuvant chemotherapy. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0297397.r002
23 Jul 2024 Decision Letter - Huei-Wen Chen, Editor Insulin-like Growth Factor 1 Receptor Expression Correlates with Programmed Death Ligand 1 Expression and Poor Survival in Non-small Cell Lung Cancer PONE-D-23-43471R1 Dear Dr. Yashiro, 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, Huei-Wen Chen, Ph.D. Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0297397.r003
Formally Accepted
26 Jul 2024 Acceptance Letter - Huei-Wen Chen, Editor PONE-D-23-43471R1 PLOS ONE Dear Dr. Yashiro, 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 If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks 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 Prof. Huei-Wen Chen Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0297397.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .