A novel genetic strategy to interrogate an unknown phenotypic modifier: Sdhc KO-Robertsonian mice develop frequent thyroid abnormalities with papillary thyroid carcinoma-like features

Jean-Pierre Bayley; Heggert G. Rebel; Peter Devilee

doi:10.1371/journal.pone.0336731

Peer Review History

Original SubmissionOctober 29, 2025
2 Feb 2026 Decision Letter - Avaniyapuram Kannan Murugan, Editor Dear Dr. Bayley, 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 Mar 19 2026 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.. 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. 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Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: This study presents a genetically elegant and technically demanding strategy combining an Sdhc knockout allele with a Robertsonian (Rb(1:7)) fusion chromosome to interrogate potential modifier effects and parent-of-origin mechanisms relevant to SDH-associated tumorigenesis. Although the model does not recapitulate PPGL, the emergence of thyroid lesions with PTC-like morphology is unexpected and scientifically interesting. Overall, the study is scientifically solid and methodologically sound, and it provides valuable data despite not fully achieving the intended tumor phenotype. I believe the manuscript is suitable for publication after the authors address the points below. This study presents a genetically elegant and technically demanding strategy combining an Sdhc knockout allele with a Robertsonian (Rb(1:7)) fusion chromosome to interrogate potential modifier effects and parent-of-origin mechanisms relevant to SDH-associated tumorigenesis. Although the model does not recapitulate PPGL, the emergence of thyroid lesions with PTC-like morphology is unexpected and scientifically interesting. Overall, the study is scientifically solid and methodologically sound, and it provides valuable data despite not fully achieving the intended tumor phenotype. I believe the manuscript is suitable for publication after the authors address the points below. Specific Comments • The central mechanistic assumption—that whole-chromosome loss contributes to the observed synergy—remains inferential. Even limited validation (FISH, CNV, allele-specific PCR) in a subset of samples would strengthen the conclusions. If additional experiments are not feasible, please explicitly acknowledge this limitation and discuss alternative explanations. • Since the study references the Hensen hypothesis and parent-of-origin effects, the manuscript should clearly state whether the Sdhc knockout allele was inherited paternally or maternally, whether reciprocal crosses were performed or considered, and how this influences interpretation. • The thyroid lesions resemble papillary thyroid carcinoma, but molecular characterization is limited. Please temper the terminology by using phrases such as “PTC-like features” or “PTC-like morphology” unless molecular markers (e.g., TTF1, PAX8, BRAF) can be provided. • A brief and balanced discussion of why this model did not generate PPGL, and how this informs future SDH-related tumor models, would improve the clarity of the Discussion section. • Minor issues include improving the resolution or annotation of some histology figures, ensuring consistent gene nomenclature for mouse vs. human, clarifying sample sizes in figure legends, and minor proofreading. A short comparison with previously published SDH mouse models would further help contextualize the novelty of the work. Reviewer #2: [1] How to perform immunohistochemistry? Description of immunohistochemistry is required (Figure 9g).[1] How to perform immunohistochemistry? Description of immunohistochemistry is required (Figure 9g). [2] Why PCR data is not available? [3] It is required to show scale bars for Figure 7 to Figure 9; magnification is not enough. [4] Current challenge would be, an establishment of mouse model to study paraganglioma, and authors partially achieved that goal. However, there is no clear statement of perspective directions. Having this work, how authors would investigate? ****** what does this mean? ). If published, this will include your full peer review and any attached files.). 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? 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NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. https://doi.org/10.1371/journal.pone.0336731.r001
Revision 1
3 Mar 2026 Author Response Reviewer #1: This study presents a genetically elegant and technically demanding strategy combining an Sdhc knockout allele with a Robertsonian (Rb(1:7)) fusion chromosome to interrogate potential modifier effects and parent-of-origin mechanisms relevant to SDH-associated tumorigenesis. Although the model does not recapitulate PPGL, the emergence of thyroid lesions with PTC-like morphology is unexpected and scientifically interesting. Overall, the study is scientifically solid and methodologically sound, and it provides valuable data despite not fully achieving the intended tumor phenotype. I believe the manuscript is suitable for publication after the authors address the points below. Specific Comments • The central mechanistic assumption—that whole-chromosome loss contributes to the observed synergy—remains inferential. Even limited validation (FISH, CNV, allele-specific PCR) in a subset of samples would strengthen the conclusions. If additional experiments are not feasible, please explicitly acknowledge this limitation and discuss alternative explanations. Reply: We refer to a mechanistic assumption underlying the Hensen Model, which is based on five publications that all showed loss of chromosome 11 in paragangliomas. These references [5-10] can be found on line 71. However, these findings do not appear relevant to the current data as we did not observe the expected phenotype. We used the word ‘synergy’ once in the manuscript, referring to thyroid abnormalities. This was based on differences in the frequency of thyroid abnormalities across cohorts and implied an interaction between the Rb chromosome and heterozygous loss of Sdhc via KO. This is not the same as chromosome loss, for which we have no evidence here. We have added the following, line 443: “We did not further explore the molecular basis of this finding, so cannot comment on whether it was associated with chromosomal loss or was a consequence of altered gene expression from metacentric Robertsonian versus telocentric chromosomes”. • Since the study references the Hensen hypothesis and parent-of-origin effects, the manuscript should clearly state whether the Sdhc knockout allele was inherited paternally or maternally, whether reciprocal crosses were performed or considered, and how this influences interpretation. Reply: We previously stated “These crosses ensured inheritance of Sdhc via either the female (4&6) or male (5&7) line in order to model human patterns of inheritance. The two cohorts carrying the paternally-transmitted Sdhc KO (4&6) simulate patients at risk of tumour development due to a paternally-inherited SDHD or SDHAF2 variant (Figure 1c)”. We have now made this more explicit from line 203 onwards. • The thyroid lesions resemble papillary thyroid carcinoma, but molecular characterization is limited. Please temper the terminology by using phrases such as “PTC-like features” or “PTC-like morphology” unless molecular markers (e.g., TTF1, PAX8, BRAF) can be provided. Reply: Indeed. We have now modified the title and text to better acknowledge this shortcoming. • A brief and balanced discussion of why this model did not generate PPGL, and how this informs future SDH-related tumor models, would improve the clarity of the Discussion section. Reply: The million-dollar question! We have now added a section to the discussion (Rodent models of PPGL) from line 398. Our guess: it’s the biochemistry. • Minor issues include improving the resolution or annotation of some histology figures, ensuring consistent gene nomenclature for mouse vs. human, clarifying sample sizes in figure legends, and minor proofreading. A short comparison with previously published SDH mouse models would further help contextualize the novelty of the work. Response: (i) “resolution or annotation of some histology figures”; Reply: Looking through the figures they appear to us of adequate or even good resolution. We have reworked most of the figures. (ii) “consistent gene nomenclature for mouse vs. human” ; Reply: we have now checked and corrected these inconsistencies. (iii) “clarifying sample sizes in figure legends” ; Reply: we have now added actual mouse numbers to figures 3, 5 and 6. (iv) “minor proofreading” ; Reply: we have carefully edited the manuscript. (v) “short comparison with previously published SDH mouse models” ; Reply: We have now added a section to the discussion (Rodent models of PPGL) from line 398. Reviewer #2: [1] How to perform immunohistochemistry? Description of immunohistochemistry is required (Figure 9g). Reply: Apologies for this oversight. A section has now been added to Methods. [2] Why PCR data is not available? Reply: The gels were of poor quality and not fit for display. We have therefore removed the sentence referring to this data. [3] It is required to show scale bars for Figure 7 to Figure 9; magnification is not enough. Reply: This has now been corrected. [4] Current challenge would be, an establishment of mouse model to study paraganglioma, and authors partially achieved that goal. However, there is no clear statement of perspective directions. Having this work, how authors would investigate? Reply: The million-dollar question! We have now added a section to the discussion (Rodent models of PPGL) from line 398. Our guess: it’s the biochemistry. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0336731.r002
10 Mar 2026 Decision Letter - Avaniyapuram Kannan Murugan, Editor A novel genetic strategy to interrogate an unknown phenotypic modifier: Sdhc KO-Robertsonian mice develop frequent thyroid abnormalities with papillary thyroid carcinoma-like features PONE-D-25-58546R1 Dear Dr. Bayley, 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. For questions related to billing, please contact and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support .. 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, Avaniyapuram Kannan Murugan, M.Phil., Ph.D. Academic Editor PLOS One Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0336731.r003
Formally Accepted
Acceptance Letter - Avaniyapuram Kannan Murugan, Editor PONE-D-25-58546R1 PLOS One Dear Dr. Bayley, 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. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. 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. Avaniyapuram Kannan Murugan Academic Editor PLOS One https://doi.org/10.1371/journal.pone.0336731.r004

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