Circulating Exosomal miR-1290 for Diagnosis of Epithelial Ovarian Cancer

Round 1

Reviewer 1 Report

In the present study, Jeon et al. examine the clinical value of miRNA-1290 and its potential target (suppressor of cytokine signaling 4, SOCS4) in epithelial ovarian cancer (EOC). The authors selected two miRNA candidates (miR-1246 and miR-1290) from serum exosomal miRNA profiling of patients with EOC. Subsequently, the authors chose miR-1290 as a candidate ovarian cancer diagnostic marker because miR-1290 expression was increased in both serum and tissue in EOC patients. In addition, the authors note that serum exosomal miR-1290 combined with CA125 has potential diagnostic power. This manuscript has some novel findings, but some points deserve further attention.

1. I wonder whether the authors want to develop a diagnostic marker for both serum and tissue. If so, the authors should evaluate enough biological samples with clinical data and emphasize this point throughout the manuscript. miR-1246 also looks promising for serum samples. What was the AUC value of miR1246, and the combination of miR1290 and miR1246? In figure 3: add n number in x axis.
2. The authors should add positive predictive value (PPV) and negative predictive value (NPV). In figure 4, the authors should include a dot plot.
3. In figure 5F, the author should include n number with percentage.
4. Ova 1 and Overa test should be mentioned in the introduction and discussion sections.
5. The authors need to expand on what is known, the mechanism of action, and potential pathways for miR-1290 (PMID: 34897783).   

 Minor

1. The authors used only 34 FFPE tissue samples for immunohistochemistry among 67 samples (Table 1). Is there a reason that only a portion of the samples was used for immunohistochemistry?
2. The authors need to check “n number” tested in each experiment.
3. The first line of the results section is not a sentence.
4. The images in figures 1 and 2 are out of focus.
5. Kwang Seock Kim is not mentioned in the Author Contributions section.
6. Reference format should be consistent.

Author Response

Thank you for your comments for this manuscripts. Thank you so much for the good point outs and comments. We have made corrections based on your comments and provided appropriate responses.

1. I wonder whether the authors want to develop a diagnostic marker for both serum and tissue. If so, the authors should evaluate enough biological samples with clinical data and emphasize this point throughout the manuscript. miR-1246 also looks promising for serum samples. What was the AUC value of miR-1246, and the combination of miR-1290 and miR-1246? In figure 3: add n number in x axis.

Response: Blood is simpler than tissue to be used for diagnosis. However, miR-1246 is not promising in serum samples (Figure 3. C and D). Therefore, miR-1246 was excluded.

2. The authors should add positive predictive value (PPV) and negative predictive value (NPV). In figure 4, the authors should include a dot plot.

Response: Already changed. Line 278 to line 282 in page 9. And the dot plot graph was added to Supplementary Figure 3.

3. In figure 5F, the author should include n number with percentage.

Response: Already include number of samples with percentage in Figure 5F.

4. Ova1 and Overa test should be mentioned in the introduction and discussion sections.

Response: sentences about OVA1 added to introduction and discussion and reference. Line 60 to line 64 in page 2. 

5. The authors need to expand on what is known, the mechanism of action, and potential pathways for miR-1290 (PMID: 34897783).

Response: sentences about this issues added to discussion shortly because this study is not for mechanism of miR-1290 but for role of biomarker detecting ovarian cancer. Line 329 to line 331 and line 366 to line 368 in page 11.

Minor

1. The authors used only 34 FFPE tissue samples for immunohistochemistry among 67 samples (Table 1). Is there a reason that only a portion of the samples was used for immunohistochemistry?

A) We also want tried to check all samples, but the amount was not enough for IHC staining and mRNA expression at the same time. Therefore, mRNA expression was first confirmed using all of the tissue samples, and IHC staining was performed with the remaining tissue.

2. The authors need to check “n number” tested in each experiment.

A) Already changed. line 154 to line 158 in page 4, and line 300 to line 302 in page 9

3. The first line of the results section is not a sentence.

1. A) Already changed. line 224 to line 225 in page 5

4. The images in figures 1 and 2 are out of focus.

A) Already changed in page 7

5. Kwang Seock Kim is not mentioned in the Author Contributions section.

A) He has contributed to the writing, review, and/or revision of the manuscript, and these are mentioned to Contributions section. (all authors contributed to the writing, review, and/or revision of the manuscript.)

6. Reference format should be consistent.

A) Already changed in page 12 to 13

Reviewer 2 Report

The manuscript by Jeon et al. examines the levels of miRNAs in serum and tissue from patients with either benign disease or ovarian cancer. Overall, the study design is appropriate and this is an interesting contribution to the field. 

Minor comments:

1. The figures appear in low resolution. They should be replaced with high resolution figures.
2. In Figure 2, the miRNA's of interest should be indicated.
3. More description of what is being tested should be included in the results sections.

Author Response

Thank you for reviewing our manuscript and for pointing out something I hadn't thought of in some part. We corrected that part.

Minor comments:

1. The figures appear in low resolution. They should be replaced with high resolution figures.

A) Already changed in page 7

2. In Figure 2, the miRNA's of interest should be indicated.

A) Already changed on Figure 2 in page 7

3. More description of what is being tested should be included in the results sections.

A) Already changed. line 246 in page 7, line 264 to line 266 in page 7, line 278 to line 279 in page 8, and line 297 to line 302 in page 9