Oxidative Stress and ROS-Mediated Signaling in Leukemia: Novel Promising Perspectives to Eradicate Chemoresistant Cells in Myeloid Leukemia
, Elena Cesaro, Rosa Catapano, Raffaele Sessa, Alessandra Lo Bianco, Paola Izzo and Michela Grosso *Round 1
Reviewer 1 Report
I reviewed with great interest the paper by Silvia Trombetti and co-workers on emerging roles of ROS in leukemia and ROS-based therapeutic approaches.
Nowadays the issue of ROS production in physiological and pathological models, and the consequence oxidative stress into the cells, is becoming more and more fascinating either in experimental than in clinical models. This issue is particularly hot in hematopoietic malignancies and overall, in myelodysplastic syndrome and leukemias. Mechanisms leading to ROS-dependent cellular death is still partially understood particularly in hematologic malignancies and is a field of intensive research even in the COVID era.
Author’s aim is to review the different findings supporting the relevance of ROS imbalance in the leukemogenic process and the crucial role of elevated ROS levels in tumor promotion and progression. Authors are focusing on the molecular mechanisms of aberrant ROS production in leukemia cells as well as on the redox-dependent signaling pathways involved in the leukemogenic process and on the importance of ROS-based therapeutically approaches
In my opinion, the purpose of the article is congruent with recent research advances in this complex subject and contributes to the dissemination of knowledge outside the narrow field of experts.
Paper is well written. References are updated and all the figures are very usefulness and well-defined
To improve the paper, I suggest the following:
Major scientific point:
The review describes ROS-dependent mechanism od cellular death and future therapeutic options (strictly dependent by ROS homeostasis) in acute leukemia, but it are often introduced concepts related to other section of hematology: chronic lymphatic leukemia, chronic myeloid leukemia and multiple myeloma. Line 141 (Ref 38), 146 (Ref 43), 186 (Ref 50, 51), line 194 (ref 56), line 201 (ref 59) , line 284 (ref 83). I believe that putting together different models/concepts could be confounding and not always helpful from the ROS homeostasis point of view. In particular the role of BCL2 in CLL is deeply different to that in acute leukemia: BCR-ABL pathway mutation characteristic of CML is present only in a subtype of acute lymphoblastic leukemia; multiple myeloma has a completely different etiopathogenetic and signaling pathways from acute leukemia. I. suggest to concentrate on Myeloid. Leukemias and MDS and cut (or simply mention) the other refers.
The paper is well written, easy to read but too long, in particular the introduction. I suggest author to summarize the introduction eliminating the concept clarified elsewhere in the text.
Discussion is definitely too long and can be significantly shortened without losing any meanings.
I suggests the implement of the figures captures with the description and explanation of mechanisms and pathway and removing the description in the text.
Minor points:
In line 346: Bortezomib is now approval only for Multiple Myeloma not AML or CLL
Line 216: Please add reference
Finally, to improve readability, I suggest to use as less abbreviations as possible
Author Response
Manuscript ID: IJMS-1036116
Reviewer: 1
- “The review describes ROS-dependent mechanism of cellular death and future therapeutic options (strictly dependent by ROS homeostasis) in acute leukemia, but it are often introduced concepts related to other section of hematology:…... I believe that putting together different models/concepts could be confounding and not always helpful from the ROS homeostasis point of view…”.
We agree with this insightful comment. We took this point and extensively revised the manuscript that, according to the reviewer’s suggestion, is now focused exclusively on the role of ROS in AML.
- “The paper is well written, easy to read but too long, in particular the introduction. I suggest author to summarize the introduction eliminating the concept clarified elsewhere in the text”
We are grateful to Reviewer 1 for this positive comment. We modified the Introduction Section by adding a brief description of AML and removing lines 40-80 that now are part of a new paragraph: “Energy metabolism and cell redox state in myeloid stem cells”.
- “Discussion is definitely too long and can be significantly shortened without losing any meanings”
We took the reviewer comment and addressed this issue by eliminating redundant or unnecessary parts throughout the manuscript.
- “I suggest the implement of the figures captures with the description and explanation of mechanisms and pathway and removing the description in the text”.
We are thankful to Reviewer 1 for this suggestion and accordingly we implemented figure captures where possible (figures 4-5).
- “In line 346: Bortezomib is now approval only for Multiple Myeloma not AML or CLL”
We are grateful to Reviewer 1 for this comment and removed this sentence from the text.
- “Line 216: Please add reference”
We took this point and added references 65, 83-85.
- “Finally, to improve readability, I suggest to use as less abbreviations as possible”
We took the reviewer’s comment and revised manuscript in order to avoid an excess of abbreviations.
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
In this review Dr. Trombetti and co-authors highlight a topic of great interest which is the role of ROS and oxidative state in the pathogenesis of leukemia and novel ROS-based therapeutic approaches in leukemia malignancies.
General comment
This is a wide and intriguing area of research that may benefit from a synthesis of the knowledge. However, the way the manuscript is currently framed is problematic.
The Introduction is not very informative and needs development. The literature overview does not seem to be as comprehensive as needed to summarize the state-of-the-art of this literature. The rationale for the review is weak and needs to be corroborated by proper justification. The objectives of the study are not spelled out at all, but must be stated.
Overall, the review is ambitious in its intentions but seems to lack a proper structure.
The comments below are meant to improve the overall quality of this submission.
Title
I question whether the title is truly descriptive of this work, possibly not.
I strongly suggest changing the current title of the manuscript since it does not describe the discussed content in the submitted review.
I know that changing the title is a radical change, but my impression is this submission is more an overview of the Reactive Oxygen Species in the contest of hematopoietic malignancy and is specifically focused on their molecular mechanisms in Leukemia.
However, not very much information is about the emerging role of novel ROS-based therapeutic approaches, thus, I wonder whether the title could be made more comprehensive.
Abstract
The Abstract is not very informative. I’d suggest summarizing at least the main findings of the review or concisely articulate on this.
Introduction and Discussion
- The “Introduction” section of the manuscript requires a revision. First of all, the authors need to expand the review of literature that is relevant to their study, references should be added, e.g line 90 “ Anyway, given their high rate of ROS production, leukemic cells have a lower buffering capacity against ROS disruption that make them more sensitive to pro-oxidant treatments than their normal counterpart ( ). Line 392 “Several mechanisms have been described as involved in aberrant ROS production in leukemia cells, including oncogene activation, mitochondrial disfunctions and metabolic changes. Furthermore, many antioxidant systems appear to be dysregulated, leading to increased overall ROS levels that allow cell survival without surpassing a deadly threshold, even under permanent… ( )
- The authors should improve the description about the relationship between antioxidant and pro-oxidant strategies as antileukemia therapeutic tools.
The paper does not really accomplish is stated goal, As a final remark for the Introduction and discussion, I wonder whether the conceptual framework of the review could be made clearer to the reader by openly stating the study questions.
My recommendation is: Major revisions
Author Response
Manuscript ID: IJMS-1036116
Reviewer: 2
- “I question whether the title is truly descriptive of this work, possibly not. I strongly suggest changing the current title of the manuscript since it does not describe the discussed content in the submitted review…” “However, not very much information is about the emerging role of novel ROS-based therapeutic approaches, thus, I wonder whether the title could be made more comprehensive”
We agree with this insightful comment. We changed the title in order to make it consistent with the revised content of the manuscript that is now focused exclusively on AML.
- “The Abstract is not very informative. I’d suggest summarizing at least the main findings of the review or concisely articulate on this.…”
We are thankful to the reviewer for this comment. We took the point and modified the Abstract that in the revised version more clearly summarizes the content of the manuscript.
- “The Introduction section of the manuscript requires a revision. First of all, the authors need to expand the review of literature that is relevant to their study, references should be added”
We took this point and extensively revised the Introduction Section and added further references (ref. 6-12, 15-19).
- “The authors should improve the description about the relationship between antioxidant and pro-oxidant strategies as antileukemia therapeutic tools”
We agree with this insightful comment and accordingly we revised the last paragraph regarding this issue by adding more details and comments (lines 442-447, 453-480, 498-506, 510-514).
Author Response File: Author Response.pdf
Reviewer 3 Report
Major point:
Related review papers are well published. Below are a few related review papers in recent years:
1. The roles of reactive oxygen species (ROS) and autophagy in the survival and death of leukemia cells. Crit Rev Oncol Hematol. 2017 Apr;112:21-30. doi: 10.1016/j.critrevonc.2017.02.004.
2. The Role of Reactive Oxygen Species in Acute Myeloid Leukaemia. Int J Mol Sci. 2019 Nov 28;20(23):6003. doi: 10.3390/ijms20236003.
3. The Cooperative Relationship between STAT5 and Reactive Oxygen Species in Leukemia: Mechanism and Therapeutic Potential. Cancers (Basel). 2018 Sep 27;10(10):359. doi: 10.3390/cancers10100359.
4. Oxidative stress in chronic lymphocytic leukemia: still a matter of debate. Leuk Lymphoma. 2019 Apr;60(4):867-875. doi: 10.1080/10428194.2018.1509317.
5. Reactive oxygen species in haematopoiesis: leukaemic cells take a walk on the wild side. J Exp Clin Cancer Res. 2018 Jun 26;37(1):125. doi: 10.1186/s13046-018-0797-0.
6. Oxidative stress in normal hematopoietic stem cells and leukemia. APMIS. 2018 Apr;126(4):284-294. doi: 10.1111/apm.12822.
Minor points:
- The order of Figures 4 and 5 should be switched.
- L25-28 is confusing: "Conversely, excessive ROS have detrimental effects caused by protein, lipid and DNA damage and subsequent disruption of cell functions [1,2] even though it is well established that a moderate increase in ROS levels promotes cancer initiation, 27 development and progression [3]."
- L30: "eustress" and L32: "oxidative distress". Why italics?
- Whether ROS plays a different role in different types of leukemia? AML, CML, ALL.....
Author Response
Manuscript ID: IJMS-1036116
Reviewer: 3
- “Related review papers are well published”.
We agree with the reviewer comment that there is an ever-growing body of literature addressing the pathogenetic significance of ROS in leukemia. However, we wish to underline that, with respect to previous published papers mainly focused on specific topics, our paper provides a more comprehensive overview of both aberrant ROS sources, signaling and energy metabolism as novel therapeutic targets to eradicate residual/chemoresistant disease in AML.
“The order of Figures 4 and 5 should be switched.
We took the reviewer point and switched the order of these two figures.
- “L25-28 is confusing: "Conversely, excessive ROS have detrimental effects caused by protein, lipid and DNA damage and subsequent disruption of cell functions [1,2] even though it is well established that a moderate increase in ROS levels promotes cancer initiation, 27 development and progression [3]."
We are grateful to the reviewer for this comment and modified the text to make the meaning of this sentence clearer (lines 30-32, 39-40).
- “L30: "eustress" and L32: "oxidative distress". Why italics?”
We took the point and amended the text typing error.
- “Whether ROS plays a different role in different types of leukemia? AML, CML, ALL....”.
We agree with the reviewer comment that the different significance of ROS in leukemia subtypes may generate confusion. According to this observation, we extensively revised the manuscript that is now exclusively focused on the role of ROS in AML.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
I reviewed with great interest the manuscript by Silvia Trombetti and co-authors, the manuscript is now considerably improved according to my suggestions and can be accepted as it is.
