PGENETICS-D-25-00882

A tumor-suppressive role of the PRC1 Polycomb epigenetic complex in the maintenance of adult Drosophila intestinal stem cell identity

PLOS Genetics

Dear Dr. Rousset,

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Giovanni Bosco, Ph.D.

Section Editor

PLOS Genetics

Giovanni Bosco

Section Editor

PLOS Genetics

Aimée Dudley

Editor-in-Chief

PLOS Genetics

Anne Goriely

Editor-in-Chief

PLOS Genetics

Journal Requirements:

1) Please ensure that the CRediT author contributions listed for every co-author are completed accurately and in full.

At this stage, the following Authors/Authors require contributions: Aurélia Joly, Ana-Maria Popmihaylova, Corinne Rancurel, Julie Soltys, Lauriane Fritsch, Rihab Loudhaief, Armel Gallet, Edan Foley, and Anne-Marie Martinez. Please ensure that the full contributions of each author are acknowledged in the "Add/Edit/Remove Authors" section of our submission form.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Authors:

Please not that one review is uploaded as an attachment.

Reviewer #1: In this manuscript, Joly et al characterise the phenotype of loss-of-function of members of the polycomb repressive complex 1 (PRC1) in the Drosophila intestine. They show that loss of PRC1 results in clusters of cells that lose intestinal markers and become neoplastic. They also perform particularly impressive serial transplant experiments to show that these cells acquire the capacity to become malignant and metastatic. This study will be of interest to understanding how epigenetic regulators can influence cancer onset.

There are a few issues that would improve the manuscript and help it achieve its fullest impact:

1. The characterisation of the PRC1-deficient cells could be expanded. While the authors show loss of esg expression and characterise proliferation, they claim that the cells lose intestinal identity. Indeed, this is in the title of the manuscript. It would be important to use several markers of intestinal and intestinal stem cell identity to support this statement, as loss of a single marker is not enough to draw strong conclusions about the identity of a cell type. Similarly, the RNAseq shows decreased expression of epithelial markers, but these are not validated in the tumours, despite the availability of many antibodies from the DSHB.

2. Secondly, some of the RNAseq findings should be validated. The authors make several claims based only on the analysis of the sequencing data, but only show that JAK/STAT signalling is activated using the 10X-STAT-GFP reporter. This would be especially helpful when arguing that the Toll and Imd pathways are activated autonomously in the tumours, but showing upregulation of Zfh1 and Chinmo would also be useful. This would increase confidence in the RNAseq dataset and the authors’ conclusions. Similarly, when discussing the upregulation of JAK/STAT independently of the ligands, it would be important to validate that ligand expression is indeed unchanged, or use more caution when discussing the results (eg p.18, lines 448-9).

3. The authors should be cautious when interpreting negative data from RNAi experiments. To conclude that Zfh1 and Chinmo are indeed not relevant to the PRC1 phenotype, it would be important to demonstrate that the two proteins are indeed knocked down by RNAi expression.

Minor comment – a typo in the legend for Fig. 3 “transcriptomique” should be “transcriptomic”.

Reviewer #2: I uploaded my comments as an attachment.

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Have all data underlying the figures and results presented in the manuscript been provided?

Large-scale datasets should be made available via a public repository as described in the PLOS Genetics  data availability policy, and numerical data that underlies graphs or summary statistics should be provided in spreadsheet form as supporting information.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Joly_PRC1 tumor_Plos G 2025.docx

Dear Dr Rousset,

We are pleased to inform you that your manuscript entitled "A tumor-suppressive role of the PRC1 Polycomb epigenetic complex in the maintenance of adult Drosophila intestinal stem cell identity" has been editorially accepted for publication in PLOS Genetics. Congratulations!

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Yours sincerely,

Giovanni Bosco, Ph.D.

Section Editor

PLOS Genetics

Giovanni Bosco

Section Editor

PLOS Genetics

Aimée Dudley

Editor-in-Chief

PLOS Genetics

Anne Goriely

Editor-in-Chief

PLOS Genetics

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Comments from the reviewers (if applicable):

Reviewer's Responses to Questions

Comments to the Authors:

Please note here if the review is uploaded as an attachment.

Reviewer #1: The authors have addressed all the comments satisfactorily. Congratulations on an interesting and thorough manuscript!

Reviewer #2: After carefully evaluating the revised manuscript and the authors’ responses to reviewers, I find that the study has been substantially improved. The authors have addressed the major concerns by including extra data, strengthening controls, and clarifying the presentation and interpretation of their results, thereby enhancing the overall rigor of the work.

In particular, the authors performed the suggested experiment to examine udp3 expression in cells surrounding the tumors, and did not observe any changes of upd3 expression either within the tumors or in the adjacent tissues. These results ruled out the involvement of JAK/STAT ligands produced by stressed ECs during ISC proliferation and further confirmed that PRC1-LOF activates JAK-STAT signaling through a direct mechanism.

Overall, this is a solid, well-executed study with clear conceptual relevance and will be of interest to the field of stem cell biology. I believe the manuscript is suitable for publication in PLOS Genetics, and no further revision is required.

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Have all data underlying the figures and results presented in the manuscript been provided?

Large-scale datasets should be made available via a public repository as described in the PLOS Genetics  data availability policy, and numerical data that underlies graphs or summary statistics should be provided in spreadsheet form as supporting information.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: Yes:  Peng Zhang

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