Differential Expression of AURKA, AURKB, and PLK1 Modulates Clinical Outcomes and Survival in Acute Myeloid Leukemia

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The presented paper is devoted to evaluation of the expression of Aurora Kinases AURKA and AURKB, and Polo-like Kinase PLK1 in bone marrow and peripheral blood of the patients diagnosed with acute myeloid leukemia as well as healthy volunteers. Authors demonstrated the significant decrease in the expression of AURKA and significant increase of PLK1 in AML patients comparing with healthy donors. Moreover, the correlation analysis demonstrated that coexpression of AURKA, AURKB and PLK1 is correlated with survival rate. The study seems to be relevant to the scope of the journal, and the amount of the presented data is sufficient for the publication. However, there several issues to be addressed:

1. The English editing should be performed. Typos and grammar issues should be eliminated.
2. Figure illustrating the role of AURKA, AURKB and PLK1 in cancer cell functioning should be added to the Introduction section.
3. The role of Aurora kinases in autophagy should be briefly described in the Introduction section.
4. Is any autophagy modulator used of could it be potentially used in the therapy of AML? Could AURKA, AURKB or PLK1 be the components of the therapeutic process? This information could be briefly added in the Discussion section.
5. Demographic data of the patients as well as inclusion/exclusion criteria should be added to the Materials and Methods section.
6. The Discussion section should be more brief and concise.
7. The primer sequences should be added.
8. Was DNAse treatment of mRNA used in PCR analysis carried out?
9. In the paragraph “Statistical analysis” the method for homogeneity evaluation as well as multiple comparison correction should be mentioned.

Comments on the Quality of English Language

The English editing should be performed. Typos and grammar issues should be eliminated.

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.

1. The English editing should be performed. Typos and grammar issues should be eliminated.

 

We thank the reviewer for pointing this out. The manuscript has been thoroughly revised to address the language issues. We have corrected all typos and grammatical errors and polished the English throughout the text. We hope the current version is much improved.

 

2. Figure illustrating the role of AURKA, AURKB and PLK1 in cancer cell functioning should be added to the Introduction section.

 

We sincerely thank the reviewer for this valuable suggestion. After careful consideration, we have decided to maintain the current structure of the manuscript without adding a new schematic figure. Our primary concern is to preserve the concise and focused nature of the Introduction, which aims to outline the key concepts and research gap without the detailed mechanistic explanation that a figure would entail. Furthermore, all figures in the manuscript are dedicated to presenting our experimental results and findings.

 

3. The role of Aurora kinases in autophagy should be briefly described in the Introduction section.

We have added a brief description of the role of Aurora kinases in autophagy to the Introduction section. We are thankful for this suggestion.

 

4. Is any autophagy modulator used of could it be potentially used in the therapy of AML? Could AURKA, AURKB or PLK1 be the components of the therapeutic process? This information could be briefly added in the Discussion section.

We sincerely appreciate your valuable suggestion. We have carefully reviewed your comment and incorporated the requested information into the Discussion section. We added information about the role of AURKA, AURKB, and PLK1 as therapeutic targets, and briefly discussed autophagic modulators, emphasizing the properties that some Aurora kinase and Polo-like kinase 1 inhibitors possess in autophagic modulation.

 

5. Demographic data of the patients as well as inclusion/exclusion criteria should be added to the Materials and Methods section.

 

The demographic data of the patient cohort and the detailed inclusion/exclusion criteria have now been added to the “Materials and Methods” sub section (4.2. Eligibility criteria) as requested.

 

6. The Discussion section should be more brief and concise.

 

We thank the reviewer for this suggestion. We have worked to make the discussion more concise. However, as we incorporated the valuable additions requested by other reviewers, a significant reduction in length was not possible without compromising the writing. The current version balances brevity with the comprehensive analysis required.

 

7. The primer sequences should be added.

We thank for this suggestion. In this study, we used commercially available, off-the-shelf TaqMan assays from Thermo Fisher Scientific. As such, the specific primer sequences are proprietary information. The assay IDs for all probes used are already provided in the methodology section, which allows for their exact identification and replication.

 

8. Was DNAse treatment of mRNA used in PCR analysis carried out?

 

We appreciate the reviewer for this important observation. Although a dedicated DNase step was not used, the TRIzol RNA extraction protocol, as stated in the manufacturer's instructions, effectively separates the genomic DNA from the RNA of interest through chloroform phase separation.

 

9. In the paragraph “Statistical analysis” the method for homogeneity evaluation as well as multiple comparison correction should be mentioned.

We have updated the “Statistical analysis” paragraph to specify the method for homogeneity evaluation using Welch's test and included Dunn's test for multiple comparisons, as suggested.

 

10. Comments on the Quality of English Language.

The English editing should be performed. Typos and grammar issues should be eliminated.

 

We fully agree with the reviewer's assessment and appreciate them highlighting this point.

 

Reviewer 2 Report

Comments and Suggestions for Authors

This study analyzed the expression of AURKA, AURKB, and PLK1 in bone marrow and peripheral blood samples from patients with acute myeloid leukemia. The authors showed that AURKA was downregulated, AURKB remained stable, and PLK1 was significantly overexpressed, independent of sex or clinical risk category. High AURKA expression in bone marrow and co-expression of all three genes were associated with better overall survival, suggesting their potential as prognostic biomarkers in AML.

1. The manuscript describes differential gene expression, but lacks a clearly stated central hypothesis or conceptual model in the Introduction. The author needs to define whether these kinases act as oncogenic drivers, prognostic biomarkers, or compensatory regulators in AML.
2. The author needs to discussthe  functional interplay of the AURKA, AURKB, and PLK1 in AML pathogenesis. For example, the author can add prior mechanistic evidence linking these kinases to AML proliferation, mitotic checkpoint dysregulation, or drug response.
3. The study uses 70 AML and 15 control samples, but statistical power and demographic comparability are not evaluated. The author can add a paragraph acknowledging this limitation and discuss how it might affect gene expression and survival analyses.
4. Figures (especially Figures 1–7) are not well described in the text and lack detailed legends regarding scales, normalization, and axis units.
5. The author can add the molecular mechanisms in the Discussion section. For example, the author can expand the discussion to explain how AURKA–PLK1 interaction, BORA-mediated activation, or Let-7 regulation could account for the observed downregulation or differential effects on survival in AML.
6. The manuscript occasionally mixes British and American spelling and contains typographical inconsistencies (e.g., commas instead of periods in p-values). Edit for consistent terminology (“hypoexpression” vs. “underexpression”) and professional tone throughout, especially in figure legends and Results summaries.

Author Response

My co-authors and I would like to thank you for your constructive feedback. Your suggestions have allowed us to refine the core ideas presented in our work. We have provided point-by-point responses to all comments below and believe the revisions have made the manuscript suitable for publication in your high-impact journal.

Kind Regards.

1. The manuscript describes differential gene expression, but lacks a clearly stated central hypothesis or conceptual model in the Introduction. The author needs to define whether these kinases act as oncogenic drivers, prognostic biomarkers, or compensatory regulators in AML.

We sincerely appreciate your valuable suggestion and have made the requested adjustments to add a central hypothesis to our work. The kinases analyzed in this study have differential expressions that reveal a possibility of their influence on the leukemogenesis process, without ruling out the physiological role of these pathways in cellular function, which may impact the survival of individuals. Furthermore, in our study we saw that PLK1 was hiperexpressed independently of sex, risk stratification, and showed no correlation with age differences, highlighting the potential of this kinase as a biomarker for the presence of the disease.

 

2. The author needs to discuss the functional interplay of the AURKA, AURKB, and PLK1 in AML pathogenesis. For example, the author can add prior mechanistic evidence linking these kinases to AML proliferation, mitotic checkpoint dysregulation, or drug response.

Dear reviewer, we would like to inform you that we have made the requested adjustments and that the suggested information is available in the "discussion" section.

 

3. The study uses 70 AML and 15 control samples, but statistical power and demographic comparability are not evaluated. The author can add a paragraph acknowledging this limitation and discuss how it might affect gene expression and survival analyses.

We appreciate the constructive suggestion and have added a paragraph to the “Materials and Methods” sub section (4.1. Biological Samples) acknowledging the imbalance between the AML and control groups and the potential impact of demographic differences that may influence the statistical power of our analyses.

 

4. Figures (especially Figures 1–7) are not well described in the text and lack detailed legends regarding scales, normalization, and axis units.

 

We thank the reviewer. All figure legends have been updated accordingly. We hope the current version is now clear.

 

5. The author can add the molecular mechanisms in the Discussion section. For example, the author can expand the discussion to explain how AURKA–PLK1 interaction, BORA-mediated activation, or Let-7 regulation could account for the observed downregulation or differential effects on survival in AML.

Thank you for your suggestion. The requested information regarding different mechanistic aspects of the kinases that are the subject of this study has been added to the discussion topic.

 

6. The manuscript occasionally mixes British and American spelling and contains typographical inconsistencies (e.g., commas instead of periods in p-values). Edit for consistent terminology (“hypoexpression” vs. “underexpression”) and professional tone throughout, especially in figure legends and Results summaries.

We understand the issue raised and appreciate your suggestions. We fully agree with the points made and have done our best to accommodate your requests in the most efficient way possible.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

No further comments