Neglected Issues in T Lymphocyte Metabolism: Purine Metabolism and Control of Nuclear Envelope Regulatory Processes. New Insights into Triggering Potential Metabolic Fragilities

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript by Torchia et. al. deals with the role of purine metabolism and nuclear envelope on the regulation of T cell immune responses. This is an interesting aspect of metabolic regulation of T cells, with broad applications in adaptive immune responses and disease progression and treatment. Overall, this work describes in detail, and as the authors state for the first time, the purinergic signaling and nuclear envelope processes governing lymphocyte function and their relevance to pathogenicity. Nevertheless, this connection is not always clear in the text. Together with the need of a more updated and relevant literature review, there are some points that need to be addressed, for the quality and clarity of this work to be improved.

 

Major points:

1.     Although the authors mention, briefly in the introduction, the connection of purine metabolism and nuclear envelope responses, this is not visible on the whole manuscript. Therefore, it appears as these two aspects are covered separately.

2.     The authors have tried to cover in detail the relevant literature on the subject in question, but it appears that some aspects could be improved (some examples are following):

·      Lines 52-57: The statements here are not backed up by relevant references.

·      Lines 59-61: Reference #8 only states the difference between P2 receptors and not their correlation to immune responses.

·      Lines 61-62: There are references missing highlighting the importance of NE in regulating innate immune responses.

·      There are studies that could be of relevance, and I couldn’t find them mentioned in the manuscript:

§  Schneider E, et al. CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression. Nat Commun. 2021.

§  Giuffrida L, et al. CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun. 2021.

§  Wang T, et al. Inosine is an alternative carbon source for CD8+-T-cell function under glucose restriction. Nat Metab. 2020.

§  Saveljeva S, et al. A purine metabolic checkpoint that prevents autoimmunity and autoinflammation. Cell Metab. 2022.

 

Minor points:

1.     In the introduction, the authors should make clearer what is the importance of T cells in the context of purine metabolism related diseases, and why it would be important to further study the purinergic system in T cells.

2.     Lines 50-54: The same sentence is repeated.

3.     Lines 57-59: the details about P2X, P2Y receptors could be omitted from introduction as they are described in detail later.

4.     Lines 106-108 need rephrasing, as it is not clear what “in different tissues within the cell” means.

5.     Line 184: change the wording to: conversion to or production of…

6.     Lines 589-592: It is not clear what is compared with what...

7.     Chapter 4.3 should be renamed from cancer cells to cancer, as the other chapters also refer to specific diseases and not cell types.

8.     Host metabolism is strongly connected to dietary uptake and studies have identified the importance of purine content in the diet for the development of diseases like gout (Choi et.al. N Engl J Med, 2004). Therefore, the authors could comment on that, maybe in the context of disease prevention or as a therapeutic approach in the conclusion.

Author Response

Dear Editor and Editorial Office

 

Thank you for the prompt and accurate editorial process. The comments have been fully taken into account. We believe that after their integration we have improved the quality of the proposed work.

Therefore, we hope that we have now done enough to revise and that we have improved the work sufficiently.

 

 

Here is a point-by-point rebuttal:

 

Rev 1

Major points:

1. Although the authors mention, briefly in the introduction, the connection of purine metabolism and nuclear envelope responses, this is not visible on the whole manuscript. Therefore, it appears as these two aspects are covered separately.
2. We concur with this assessment and are grateful to the reviewer for his/her insightful opinion. It should be noted, however, that the title makes it clear that the purpose of this review is not to definitively link the two topics, but rather to address the broader biology of lymphocytes and to examine two areas that have been insufficiently researched with respect to their importance, both from a basic biological standpoint and as a potential therapeutic avenue. Indeed, our proposal is supported by a body of evidence from existing literature and our own experimental findings. These suggest a greater degree of interconnectedness between the two subjects than was previously assumed, prompting us to put forth a number of potential biological interactions, including those between purine metabolism and the regulation of the nuclear envelope and nuclear transport. In response to the objections raised by the referee, we have added in several points along the manuscript additional elements that could account for the observed interconnection between the two themes.
3. The authors have tried to cover in detail the relevant literature on the subject in question, but it appears that some aspects could be improved (some examples are following):

• Lines 52-57: The statements here are not backed up by relevant references.
• Lines 59-61: Reference #8 only states the difference between P2 receptors and not their correlation to immune responses.
• Lines 61-62: There are references missing highlighting the importance of NE in regulating innate immune responses.
• There are studies that could be of relevance, and I couldn’t find them mentioned in the manuscript:
• Schneider E, et al. CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression. Nat Commun. 2021.
• Giuffrida L, et al. CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun. 2021.
• Wang T, et al. Inosine is an alternative carbon source for CD8+-T-cell function under glucose restriction. Nat Metab. 2020.
• Saveljeva S, et al. A purine metabolic checkpoint that prevents autoimmunity and autoinflammation. Cell Metab. 2022.

1. I am indebted to this referee for the comprehensive nature of his analysis, which enabled us to rectify and modify several erroneous elements, while supplementing our initial revision work with accurate and pertinent citations.

I would like to reassure you that the suggested corrections have been implemented and that the additions to the literature have been correctly incorporated and contextualised.

                      

 

 

 

 

Minor points:

1. In the introduction, the authors should make clearer what is the importance of T cells in the context of purine metabolism related diseases, and why it would be important to further study the purinergic system in T cells.
2. Thanks for this comment. Some sentences have been added in the introduction to emphasize the importance of T cells in the context of purine metabolism diseases, as the reviewer has suggested.
3. Lines 50-54: The same sentence is repeated.  

 

1. thank you very much the repeated sentence has now been deleted

 

2. Lines 57-59: the details about P2X, P2Y receptors could be omitted from introduction as they are described in detail later.
3. thank you very much these details have now been omitted
4. Lines 106-108 need rephrasing, as it is not clear what “in different tissues within the cell” means.
5. Line 184: change the wording to: conversion to or production of…
6. Lines 589-592: It is not clear what is compared with what...
7. Chapter 4.3 should be renamed from cancer cells to cancer, as the other chapters also refer to specific diseases and not cell types.

 

1. We apologize for these errors. Points 4 to 7 highlighted by this referee have now been appropriately amended

 

8. Host metabolism is strongly connected to dietary uptake and studies have identified the importance of purine content in the diet for the development of diseases like gout (Choi et.al. N Engl J Med, 2004). Therefore, the authors could comment on that, maybe in the context of disease prevention or as a therapeutic approach in the conclusion.

 

1. We would like to thank the referee for this excellent food for thought. It has now been briefly commented on in the text and the suggested quotation has been appropriately incorporated.

 

 

Rev2

1. Please review page 2, lines 50-54, to eliminate redundant sentences or phrases. Consolidating similar ideas into a single, cohesive sentence would improve clarity and conciseness.
2. I would like to apologize for this error and thank the referee for pointing it out. I assure you that the redundancies have been eliminated. The concepts are now consistent.

 

2. In line 106, consider rephrasing for clarity, for example: "In different tissues, intracellular concentrations of GTP vary…"
3. Thank you very much the concept has now been clarified
4. Consider addressing these errors for accuracy: In Table 1, please note that the APRT enzyme catalyzes the conversion of adenine to AMP, not AMP to adenine. Similarly, HPRT catalyzes the conversion of guanine and xanthine to their respective monophosphates.
5. I apologize for these errors and for reversing the terms of the reactions. The highlighted elements have now been corrected

 

 

 

 

Thank you for the attention that you will dedicate to our manuscript. Looking forward to hearing from you, please accept our best regards. Sincerely,

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

In this review, Naomi Torchia et al. explore the role of purine metabolism in regulating various cellular processes, with a particular focus on T-lymphocytes and nuclear envelope regulation. They provide a comprehensive summary of recent insights into the metabolic and molecular control of nuclear envelope dynamics in T-lymphocytes, highlighting its therapeutic potential. The authors thoroughly examine literature on purine balance, receptor activity, nuclear envelope reorganization, and nuclear DNA transport to the cytosol in T-lymphocytes during activation, addressing both physiological and pathological contexts. In conclusion, they propose purinergic signaling as a potential prognostic biomarker in diverse diseases, including cancer, autoimmune disorders, and viral infections like HIV.

Overall, this well-structured and engaging review is valuable for the immunology community, particularly given its insights into purine metabolism’s role in chromatin and nuclear envelope dynamics during T-cell activation and other immune functions. The review critically and constructively discusses purine signaling’s regulatory influence on immune responses, such as its dual role in promoting pro-inflammatory (danger signals) or anti-inflammatory (adenosine) functions, nuclear decondensation, and the release of nuclear DNA into the cytosol during T-cell activation—a recent advancement in the field.

This review captures the majority of state-of-the-art research and appropriately cites relevant studies. Tables and figures are generally well-presented, with some suggestions noted below.

Comments to the Authors:

1. Please review page 2, lines 50-54, to eliminate redundant sentences or phrases. Consolidating similar ideas into a single, cohesive sentence would improve clarity and conciseness.

2. In line 106, consider rephrasing for clarity, for example: "In different tissues, intracellular concentrations of GTP vary…"

3. Consider addressing these errors for accuracy: In Table 1, please note that the APRT enzyme catalyzes the conversion of adenine to AMP, not AMP to adenine. Similarly, HPRT catalyzes the conversion of guanine and xanthine to their respective monophosphates.

Author Response

Dear Editor and Editorial Office

 

Thank you for the prompt and accurate editorial process. The comments have been fully taken into account. We believe that after their integration we have improved the quality of the proposed work.

Therefore, we hope that we have now done enough to revise and that we have improved the work sufficiently.

 

 

Here is a point-by-point rebuttal:

 

Rev 1

Major points:

1. Although the authors mention, briefly in the introduction, the connection of purine metabolism and nuclear envelope responses, this is not visible on the whole manuscript. Therefore, it appears as these two aspects are covered separately.
2. We concur with this assessment and are grateful to the reviewer for his/her insightful opinion. It should be noted, however, that the title makes it clear that the purpose of this review is not to definitively link the two topics, but rather to address the broader biology of lymphocytes and to examine two areas that have been insufficiently researched with respect to their importance, both from a basic biological standpoint and as a potential therapeutic avenue. Indeed, our proposal is supported by a body of evidence from existing literature and our own experimental findings. These suggest a greater degree of interconnectedness between the two subjects than was previously assumed, prompting us to put forth a number of potential biological interactions, including those between purine metabolism and the regulation of the nuclear envelope and nuclear transport. In response to the objections raised by the referee, we have added in several points along the manuscript additional elements that could account for the observed interconnection between the two themes.
3. The authors have tried to cover in detail the relevant literature on the subject in question, but it appears that some aspects could be improved (some examples are following):

• Lines 52-57: The statements here are not backed up by relevant references.
• Lines 59-61: Reference #8 only states the difference between P2 receptors and not their correlation to immune responses.
• Lines 61-62: There are references missing highlighting the importance of NE in regulating innate immune responses.
• There are studies that could be of relevance, and I couldn’t find them mentioned in the manuscript:
• Schneider E, et al. CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression. Nat Commun. 2021.
• Giuffrida L, et al. CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun. 2021.
• Wang T, et al. Inosine is an alternative carbon source for CD8+-T-cell function under glucose restriction. Nat Metab. 2020.
• Saveljeva S, et al. A purine metabolic checkpoint that prevents autoimmunity and autoinflammation. Cell Metab. 2022.

1. I am indebted to this referee for the comprehensive nature of his analysis, which enabled us to rectify and modify several erroneous elements, while supplementing our initial revision work with accurate and pertinent citations.

I would like to reassure you that the suggested corrections have been implemented and that the additions to the literature have been correctly incorporated and contextualised.

                      

 

 

 

 

Minor points:

1. In the introduction, the authors should make clearer what is the importance of T cells in the context of purine metabolism related diseases, and why it would be important to further study the purinergic system in T cells.
2. Thanks for this comment. Some sentences have been added in the introduction to emphasize the importance of T cells in the context of purine metabolism diseases, as the reviewer has suggested.
3. Lines 50-54: The same sentence is repeated.  

 

1. thank you very much the repeated sentence has now been deleted

 

2. Lines 57-59: the details about P2X, P2Y receptors could be omitted from introduction as they are described in detail later.
3. thank you very much these details have now been omitted
4. Lines 106-108 need rephrasing, as it is not clear what “in different tissues within the cell” means.
5. Line 184: change the wording to: conversion to or production of…
6. Lines 589-592: It is not clear what is compared with what...
7. Chapter 4.3 should be renamed from cancer cells to cancer, as the other chapters also refer to specific diseases and not cell types.

 

1. We apologize for these errors. Points 4 to 7 highlighted by this referee have now been appropriately amended

 

8. Host metabolism is strongly connected to dietary uptake and studies have identified the importance of purine content in the diet for the development of diseases like gout (Choi et.al. N Engl J Med, 2004). Therefore, the authors could comment on that, maybe in the context of disease prevention or as a therapeutic approach in the conclusion.

 

1. We would like to thank the referee for this excellent food for thought. It has now been briefly commented on in the text and the suggested quotation has been appropriately incorporated.

 

 

Rev2

1. Please review page 2, lines 50-54, to eliminate redundant sentences or phrases. Consolidating similar ideas into a single, cohesive sentence would improve clarity and conciseness.
2. I would like to apologize for this error and thank the referee for pointing it out. I assure you that the redundancies have been eliminated. The concepts are now consistent.

 

2. In line 106, consider rephrasing for clarity, for example: "In different tissues, intracellular concentrations of GTP vary…"
3. Thank you very much the concept has now been clarified
4. Consider addressing these errors for accuracy: In Table 1, please note that the APRT enzyme catalyzes the conversion of adenine to AMP, not AMP to adenine. Similarly, HPRT catalyzes the conversion of guanine and xanthine to their respective monophosphates.
5. I apologize for these errors and for reversing the terms of the reactions. The highlighted elements have now been corrected

 

 

 

 

Thank you for the attention that you will dedicate to our manuscript. Looking forward to hearing from you, please accept our best regards. Sincerely,

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors did a great job addressing all the comments.

Reviewer 2 Report

Comments and Suggestions for Authors

In the revised manuscript the authors' addressed the points raised in my initial review report. The revised manuscript may be considered for publication.