PONE-D-21-10916

Intravesical macrophage migration inhibitory factor (MIF) receptors mediate bladder pain

PLOS ONE

Dear Dr. Vera,

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.

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ACADEMIC EDITOR:

Specific point 3.) (Ibudilast) as mentioned by Reviewer #1 under 5.) Review Comments to the Author may be either addressed by experiments or expanded discussion.

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PLOS ONE

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Additional Editor Comments (if provided):

Comments Academic Editor

In addition to the comments provided by the referees, please consider creating a summarizing scheme/cartoon that sums up the "induction - vesicle - MIF - receptor - HMGB1 pathway". This would greatly help readers from related fields.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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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

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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5. Review Comments to the Author

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Reviewer #1: In this manuscript (PONE-D-21-10916) Ye, Ma, et al. utilize a series of pharmacological reagents (mainly inhibitors) to demonstrate the role of MIF signaling and its associated receptors in mouse models of bladder pain. This is a great follow up to the work of the Vera laboratory and now reveals a role for CD74 in MIF-mediated bladder pain, as well as confirming previous results for CXCR4. The manuscript is generally well presented and sets up the next set of translational studies well. With that said, there are some opportunity to improve the clarity of the message and robustness of the data presented.

1. The title is a bit vague and does not capture the new role for CD74. I would recommend being clear about this as it will get readers attention easier.

2. The abstract also should be revised to include this key finding, as well as the in vivo functional observations (which are not clearly articulated).

3. Given the use of ibudilast (which blocks among other interactions, CD74/MIF) in multiple clinical settings, it would be very useful to use it in these studies. While it may be out of the scope of these studies, this may also be a good use of the CD74 KO mice (developed by one of the co-authors, Dr. Bucala).

4. The data in Table 3 are useful, but would be worth presented as a series of graphs. The authors should use their discretion as to how many conditions they show as the rest can be included as supplemental data (in the form of a table).

5. The immune changes presented are generally qualitative in terms of histology. It would be useful to quantify these, as well as other markers of inflammation via IHC. While it may end up being a negative result (based on the previous Vera laboratory report (PMID 31289792), it would still be worth assessment with appropriate quantification.

6. The qPCR data on IL1-b and TNF-a, while negative, should still be graphed in some manner (as a supplemental figure is sufficient).

7. In the conclusions, the authors should be more precise about “TLR4” as it is suggested that this is only receptor for HMGB1 (which can bind to other receptors, including TLR2 and RAGE). It may be worth just switching “TLR4” with “HMGB1” receptors or ending the sentence as such: …HMGB1 to result in bladder pain.”

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Intravesical CD74 and CXCR4, macrophage migration

inhibitory factor (MIF) receptors, mediate bladder pain

PONE-D-21-10916R1

Dear Dr. Vera,

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.

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Kind regards,

Jürgen Bernhagen

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Authors have addressed the concerns raised.

Reviewers' comments:

N/A