MicroRNA 320a and Membrane Antigens as Tools to Evaluate the Pathophysiology of Platelets Stored in Blood Banks

Round 1

Reviewer 1 Report

The study of P.C.M. Vieira et al. is devoted to finding correlations between platelet miRNA content and platelet functionality in platelet concentrates. The authors demonstrate that miR-320 and miR-127 correlated/anticorrelated with platelet P-selectin and GPIIb-IIIa expression during storage and thus could be used as indicators of PC’s quality. Although the results of the study mostly repeats the results obtained previously in the same and other groups, the large amount of samples used in this study and the performed correlation analysis makes it worth publishing in the CIMB after a major revision of the data presentation.

Major points:

1. The Introduction requires some additional information both on miRNA content in PCs (for ex., doi: 10.1080/09537104.2022.2042233) and on platelet activation during storage (for ex. https://doi.org/10.1111/vox.12364).
2. The authors test presence of four antigens on platelet surface: P2Y12, P-selectin, and GPIIb-IIIa, however, the choice of antigens was not discussed in the introduction (apart from P-selectin). Additionally, it is not clear why the P2Y12-regulating miR (miR-223) was not assessed in this study.
3. The description of flow-cytometry method lacks some important information: whether platelets were fixed, what concentrations of antibodies were used etc.
4. In Table 1 the units for membrane protein expression and miRNA expression should be given.
5. The text in Page 7, lines 3-8, is in Hispanic.
6. It should be underlined, that the data on P-selectin expression repeats most of the previously published data.
7. It seems that the data on miRNA expression during storage repeats the previously published by the same group data (doi:10.1371/journal.pone.0129399).
8. The Discussion section is very long compared to the Results section and it resembles a literature review more than a Discussion of the obtained data. I suggest the authors to transfer some of the data into the Introduction and to remove the unrelated data from the Discussion.
9. However, one point should be discussed in more detail. The P-selectin and integrin expressions are not necessary correlated with platelet functioning. There are some data that platelet functional responses do not correlate with platelet pre-activation (https://doi.org/10.1111/vox.12364).
10. The quality of the Figures is very poor.

Author Response

Dear reviewer, my co-authors and I would like to thank you for the suggestions made during this high-quality review and then we present the answers to the questions.

 

We inform that with the reviews and suggestions, we were able to improve the idea presented by our work and we appreciate the opportunity. We hope this review has left the article suitable for publication in this high-impact journal and respect in the area.

 

Kind Regards.

 

 

REVIEWER 1

 

The study of P.C.M. Vieira et al. is devoted to finding correlations between platelet miRNA content and platelet functionality in platelet concentrates. The authors demonstrate that miR-320 and miR-127 correlated/anticorrelated with platelet P-selectin and GPIIb-IIIa expression during storage and thus could be used as indicators of PC’s quality. Although the result of the study mostly repeats the results obtained previously in the same and other groups, the large amount of samples used in this study and the performed correlation analysis makes it worth publishing in the CIMB after a major revision of the data presentation.

Major points:

1. The Introduction requires some additional information both on miRNA content in PCs (for ex., doi: 10.1080/09537104.2022.2042233) and on platelet activation during storage (for ex. https://doi.org/10.1111/vox.12364).

 

R = We thank the referee for the indications of the two important articles. We have introduced their information in the introduction.

 

1. The authors test presence of four antigens on platelet surface: P2Y12, P-selectin, and GPIIb-IIIa, however, the choice of antigens was not discussed in the introduction (apart from P-selectin). Additionally, it is not clear why the P2Y12-regulating miR (miR-223) was not assessed in this study.

R = The reviewer is right, and we justify the choices of the biomarkers studied in the introduction of the manuscript. We also justify the non-inclusion of miR-223 and its important role in regulating the production of the P2Y12 membrane antigen.

 

1. The description of flow-cytometry method lacks some important information: whether platelets were fixed, what concentrations of antibodies were used etc.

 

R = The use, type and concentration of fixative for platelet fixation are controversial. For this reason, we immediately process the unfixed PC samples, after collection each day of storage, because in this way we would theoretically have a lower in vitro activation. We added this information, along with the antibody concentration and dilution, to the material and methods.

 

1. In Table 1 the units for membrane protein expression and miRNA expression should be given.

R = They were inserted in the legend of Table 1.

 

1. The text in Page 7, lines 3-8, is in Hispanic.

R= We are sorry about that, and the correction was made

1. It should be underlined, that the data on P-selectin expression repeats most of the previously published data.

 

R = We highlighted that data on the discussion.

 

1. It seems that the data on miRNA expression during storage repeats the previously published by the same group data (doi:10.1371/journal.pone.0129399).

 

R = We would like to point out that the analyzes were not performed on the same PC concentrates and that the present work aims to validate the data presented by our research group in Pontes et al., 2015 in a large number of samples. This we make clear in the introduction to the manuscript.

 

1. The Discussion section is very long compared to the Results section and it resembles a literature review more than a Discussion of the obtained data. I suggest the authors to transfer some of the data into the Introduction and to remove the unrelated data from the Discussion.

 

R = Alterations done.

 

1. However, one point should be discussed in more detail. The P-selectin and integrin expressions are not necessary correlated with platelet functioning. There are some data that platelet functional responses do not correlate with platelet pre-activation (https://doi.org/10.1111/vox.12364).

 

R = We've added more content to the suggested discussion.

 

1. The quality of the Figures is very poor.

R = We have improved their quality.

 

Reviewer 2 Report

To:

Editorial Board

Current Issues in Molecular Biology

 

 

Title: “MicroRNA 320a and membrane antigens as tools to evaluate the pathophysiology of platelets stored in blood banks”

 

 

Dear Editor,

I read this paper and I think that:

• The English of the paper should be revised by a native English speaker. Please provide.
• There are parts of the paper in Portuguese.
• The paper is difficult to be followed as it is poorly understandable.

Author Response

 

Dear reviewer, my co-authors and I would like to thank you for the suggestions made during this high-quality review and then we present the answers to the questions.

 

We inform that with the language reviews, we were able to improve the idea presented by our work and we appreciate the opportunity. We hope this review has left the article suitable for publication in this high-impact journal and respect in the area.

 

Kind Regards.

Round 2

Reviewer 1 Report

The authors have sufficiently responded to my questions, I do not have further comments.

Reviewer 2 Report

the authors well revised the paper which is now well written.