HIV Infection, Neurotoxicity, Inflammation, Premature Aging, and Therapeutic Challenges to PLWH: An Overview
, Ulhas Naik 1, Kratika Tyagi 1, Madhulika Sharma 2, Gagan Kaushal 3, Alok Bhushan 3, Michael Bukrinsky 4 and Priya Tyagi 5Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe current work mainly reports associations; however, the authors should further explore the possibility of causal relationships rather than correlation alone. Where feasible, the Discussion should outline concrete causal-inference strategies, for example approaches based on Mendelian randomization (MR) and related sensitivity analyses, as illustrated in prior studies (PMID: 38438963).
Author Response
Dear Editor,
We thank you for the opportunity to revise our manuscript. We are pleased to submit the revised version and sincerely appreciate the time and effort you and the reviewers devoted to evaluating our work. The reviewers’ thoughtful and constructive comments have significantly strengthened the manuscript. We have carefully addressed all concerns raised by the reviewers.
In the revised version, changes have been incorporated throughout the manuscript and are highlighted using track changes for ease of review.
We have attached both
1) Highlighted using track changes for ease of review.
2) Updated clean manuscript with all the changes accepted.
All authors have read and approved the revised manuscript. Should you have any questions or require additional information, please do not hesitate to contact us.
Below, we provide point-by-point responses to each reviewer’s comments. We thank you again for your consideration of our work and look forward to your response.
Sincerely,
Mudit Tyagi
On behalf of all authors
Response to the reviewers
Rev – 1:
Open Review
Quality of English Language
(x) The English could be improved to more clearly express the research.
( ) The English is fine and does not require any improvement.
Response: We have improved the language throughout the manuscript with particular attention to clarity, precision, and readability. In addition, we utilized a professional English-language editing service provided by our institution to further refine the manuscript.
Comments and Suggestions for Authors
The current work mainly reports associations; however, the authors should further explore the possibility of causal relationships rather than correlation alone. Where feasible, the Discussion should outline concrete causal-inference strategies, for example approaches based on Mendelian randomization (MR) and related sensitivity analyses, as illustrated in prior studies (PMID: 38438963).
Response: We thank the reviewer for this insightful suggestion. As recommended, we have expanded the Discussion to incorporate causal-inference frameworks, including Mendelian randomization and complementary sensitivity analyses, and have cited relevant prior work to strengthen mechanistic interpretation beyond association.
Rev2: Comments and Suggestions for Authors:
Generally, this was a well-written review. I have a number of comments that would strengthen it and make it even more impactful.
There was much redundancy and re-stating of various points in multiple places. The length of the review could be reduced once fixed and this would provide them the space to add important issues that they failed to address.
Response: We appreciate the reviewer’s positive assessment and constructive suggestions. We carefully revised the manuscript to reduce redundancy where possible. In select instances, limited repetition was retained to preserve conceptual continuity across sections addressing interconnected mechanisms.
It would be helpful to the reader to address mechanism(s) by which the acting anti-retrovirals are able to be “long acting”. Intra-muscular injection is only a small part of that issue.
Response: We fully agree and have expanded the LA-ART section to describe the pharmacokinetic and cellular mechanisms underlying long-acting efficacy, including tissue sequestration, intracellular retention, and metabolic stability.
They state (often) that reactive oxygen species have been implicated in HAND and the neurotoxicity associated with HIV-1 proteins and at least some ARTs. However, it is not just reactive species. The entire reactive species interactome is involved and some mention of this is warranted.
Response: We thank the reviewer for this important clarification. A new section has been added describing the broader reactive species interactome, including reactive nitrogen and lipid species, and their collective contribution to neurotoxicity and HAND.
They state (often) that mitochondria are involved. This is true. However, a vast literature has now been published showing that lysosome effects are upstream of HIV protein and ART induced effects on mitochondria. Thus, targeting mitochondria without also targeting upstream lysosomes will likely continue to end in failure to protect. This must be addressed in multiple places in the review but especially in section 9.6 “Mitochondrial dysfunction”.
Response: We appreciate this insightful recommendation. We have now incorporated lysosomal dysfunction as an upstream driver of mitochondrial injury at multiple points throughout the manuscript, with particular emphasis in Section 9.6.
They must address ART-induced neurotoxicity. Issues to be included should be which ART drugs, what mechanisms have been implicated, and which cell types have been shown to be involved.
Response: We have expanded this section to further detail ART-associated neurotoxicity, focusing primarily on glial cells, consistent with the central theme of the manuscript.
On line 471 they state that “One of the most detrimental consequences…” This is unknown. It is clearly detrimental and implicated but the relative importance is unknown.
Response: Thank you for noting this. We have revised the wording to reflect uncertainty regarding relative contribution.
On line 519 I think the 2 citations are incorrect. They must cite the very important work of Kelly Jordan-Sciutto.
Response: We have now incorporated the recommended seminal studies. Due to revisions, the originally cited line numbers have shifted accordingly.
On line 619 it is odd that the seminal paper on Tat levels in the CNS of at least a subset of patients on ART was not cited; Tory Johnson PNAS.
Response: Thanks a lot for the suggestion, we have now included the suggested references from Tory Johnson PNAS.
On line 641 the citations should be numbered.
Response: We appreciate the meticulous reading of the manuscript by the reviewer; We have rectified our mistake.
On line 648 and line 812 they mention lysosome dysfunction and then cite papers that have little to do about this issue. Effects of HIV-1 proteins on lysosomes and the greater lysosomal system are available and must be cited.
Response: We thank the reviewer for the careful reading; we have now improved the text and incorporated several relevant references.
In section 9.8 “Structural changes” they should discuss subcellular morphological changes especially to lysosomes and mitochondria.
Response: We thank the reviewer for the important comment. Besides mentioning structural changes to lysosomes and mitochondria at several places in the text; now we have also included a dedicated paragraph describing morphological changes to lysosomes and mitochondria.
In section 10.2 they should include information about targeting the greater lysosomal system. A number of companies have made great strides in this regard.
Response: We appreciate the reviewer’s insightful suggestion. We have now expanded this section to include emerging therapeutic approaches targeting lysosomal pathways.
Once again, we sincerely thank the reviewers for their careful evaluation and valuable recommendations, which have substantially improved the clarity, comprehensiveness, and impact of our manuscript.
Author Response File: 
Author Response.docx
Reviewer 2 Report
Comments and Suggestions for AuthorsGenerally, this was a well-written review. I have a number of comments that would strengthen it and make it even more impactful.
There was much redundancy and re-stating of various points in multiple places. The length of the review could be reduced once fixed and this would provide them the space to add important issues that they failed to address.
It would be helpful to the reader to address mechanism(s) by which the long acting anti-retrovirals are able to be “long acting”. Intra-muscular injection is only a small part of that issue.
They state (often) that reactive oxygen species have been implicated in HAND and the neurotoxicity associated with HIV-1 proteins and at least some ARTs. However, it is not just reactive species. The entire reactive species interactome is involved and some mention of this is warranted.
They state (often) that mitochondria are involved. This is true. However, a vast literature has now been published showing that lysosome effects are upstream of HIV-1 protein and ART induced effects on mitochondria. Thus, targeting mitochondria without also targeting upstream lysosomes will likely continue to end in failure to protect. This must be addressed in multiple places in the review but especially in section 9.6 “Mitochondrial dysfunction”.
They must address ART-induced neurotoxicity. Issues to be included should be which ART drugs, what mechanisms have been implicated, and which cell types have been shown to be involved.
On line 471 they state that “One of the most detrimental consequences…” This is unknown. It is clearly detrimental and implicated but the relative importance is unknown.
On line 519 I think the 2 citations are incorrect. They must cite the very important work of Kelly Jordan-Sciutto.
On line 619 it is odd that the seminal paper on Tat levels in the CNS of at least a subset of patients on ART was not cited; Tory Johnson PNAS.
On line 641 the citations should be numbered.
On line 648 and line 812 they mention lysosome dysfunction and then cite papers that have little to do about this issue. Effects of HIV-1 proteins on lysosomes and the greater lysosomal system are available and must be cited.
In section 9.8 “Structural changes” they should discuss subcellular morphological changes especially to lysosomes and mitochondria.
In section 10.2 they should include information about targeting the greater lysosomal system. A number of companies have made great strides in this regard.
Author Response
Dear Editor,
We thank you for the opportunity to revise our manuscript. We are pleased to submit the revised version and sincerely appreciate the time and effort you and the reviewers devoted to evaluating our work. The reviewers’ thoughtful and constructive comments have significantly strengthened the manuscript. We have carefully addressed all concerns raised by the reviewers.
In the revised version, changes have been incorporated throughout the manuscript and are highlighted using track changes for ease of review.
We have attached both
1) Highlighted using track changes for ease of review.
2) Updated clean manuscript with all the changes accepted.
All authors have read and approved the revised manuscript. Should you have any questions or require additional information, please do not hesitate to contact us.
Below, we provide point-by-point responses to each reviewer’s comments. We thank you again for your consideration of our work and look forward to your response.
Sincerely,
Mudit Tyagi
On behalf of all authors
Response to the reviewers
Rev – 1:
Open Review
Quality of English Language
(x) The English could be improved to more clearly express the research.
( ) The English is fine and does not require any improvement.
Response: We have improved the language throughout the manuscript with particular attention to clarity, precision, and readability. In addition, we utilized a professional English-language editing service provided by our institution to further refine the manuscript.
Comments and Suggestions for Authors
The current work mainly reports associations; however, the authors should further explore the possibility of causal relationships rather than correlation alone. Where feasible, the Discussion should outline concrete causal-inference strategies, for example approaches based on Mendelian randomization (MR) and related sensitivity analyses, as illustrated in prior studies (PMID: 38438963).
Response: We thank the reviewer for this insightful suggestion. As recommended, we have expanded the Discussion to incorporate causal-inference frameworks, including Mendelian randomization and complementary sensitivity analyses, and have cited relevant prior work to strengthen mechanistic interpretation beyond association.
Rev2: Comments and Suggestions for Authors:
Generally, this was a well-written review. I have a number of comments that would strengthen it and make it even more impactful.
There was much redundancy and re-stating of various points in multiple places. The length of the review could be reduced once fixed and this would provide them the space to add important issues that they failed to address.
Response: We appreciate the reviewer’s positive assessment and constructive suggestions. We carefully revised the manuscript to reduce redundancy where possible. In select instances, limited repetition was retained to preserve conceptual continuity across sections addressing interconnected mechanisms.
It would be helpful to the reader to address mechanism(s) by which the acting anti-retrovirals are able to be “long acting”. Intra-muscular injection is only a small part of that issue.
Response: We fully agree and have expanded the LA-ART section to describe the pharmacokinetic and cellular mechanisms underlying long-acting efficacy, including tissue sequestration, intracellular retention, and metabolic stability.
They state (often) that reactive oxygen species have been implicated in HAND and the neurotoxicity associated with HIV-1 proteins and at least some ARTs. However, it is not just reactive species. The entire reactive species interactome is involved and some mention of this is warranted.
Response: We thank the reviewer for this important clarification. A new section has been added describing the broader reactive species interactome, including reactive nitrogen and lipid species, and their collective contribution to neurotoxicity and HAND.
They state (often) that mitochondria are involved. This is true. However, a vast literature has now been published showing that lysosome effects are upstream of HIV protein and ART induced effects on mitochondria. Thus, targeting mitochondria without also targeting upstream lysosomes will likely continue to end in failure to protect. This must be addressed in multiple places in the review but especially in section 9.6 “Mitochondrial dysfunction”.
Response: We appreciate this insightful recommendation. We have now incorporated lysosomal dysfunction as an upstream driver of mitochondrial injury at multiple points throughout the manuscript, with particular emphasis in Section 9.6.
They must address ART-induced neurotoxicity. Issues to be included should be which ART drugs, what mechanisms have been implicated, and which cell types have been shown to be involved.
Response: We have expanded this section to further detail ART-associated neurotoxicity, focusing primarily on glial cells, consistent with the central theme of the manuscript.
On line 471 they state that “One of the most detrimental consequences…” This is unknown. It is clearly detrimental and implicated but the relative importance is unknown.
Response: Thank you for noting this. We have revised the wording to reflect uncertainty regarding relative contribution.
On line 519 I think the 2 citations are incorrect. They must cite the very important work of Kelly Jordan-Sciutto.
Response: We have now incorporated the recommended seminal studies. Due to revisions, the originally cited line numbers have shifted accordingly.
On line 619 it is odd that the seminal paper on Tat levels in the CNS of at least a subset of patients on ART was not cited; Tory Johnson PNAS.
Response: Thanks a lot for the suggestion, we have now included the suggested references from Tory Johnson PNAS.
On line 641 the citations should be numbered.
Response: We appreciate the meticulous reading of the manuscript by the reviewer; We have rectified our mistake.
On line 648 and line 812 they mention lysosome dysfunction and then cite papers that have little to do about this issue. Effects of HIV-1 proteins on lysosomes and the greater lysosomal system are available and must be cited.
Response: We thank the reviewer for the careful reading; we have now improved the text and incorporated several relevant references.
In section 9.8 “Structural changes” they should discuss subcellular morphological changes especially to lysosomes and mitochondria.
Response: We thank the reviewer for the important comment. Besides mentioning structural changes to lysosomes and mitochondria at several places in the text; now we have also included a dedicated paragraph describing morphological changes to lysosomes and mitochondria.
In section 10.2 they should include information about targeting the greater lysosomal system. A number of companies have made great strides in this regard.
Response: We appreciate the reviewer’s insightful suggestion. We have now expanded this section to include emerging therapeutic approaches targeting lysosomal pathways.
Once again, we sincerely thank the reviewers for their careful evaluation and valuable recommendations, which have substantially improved the clarity, comprehensiveness, and impact of our manuscript.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThere were some issues that not be solved.
Author Response
Response to the Reviewers:
We have improved the language throughout the manuscript with particular attention to clarity, precision, and readability using a professional English-language editing service provided by our institution. However, to further ensure the highest linguistic and editorial standards and to maximize the impact and clarity of the work, we would greatly appreciate the assistance of the journal’s expert language editors, if needed.
Line 802 "Briefly," followed by multiple paragraphs! This is not correct use of the English language.
Response: We appreciate the reviewer’s insightful suggestion. We have now revised the wording.
lines 995-996 They absolutely must cite the seminal literature by the J.D. Geiger lab showing the lysosome dysfunction precedes and amplifies mitochondrial damage. The following sentence has citations (22, 160, 179-184) that are at best questionable with regard to the content of the sentence.
Response: As suggested, we have now rectified. We have now included some more relevant references, including seminal references from Dr. J.D. Geiger lab (precedes and amplifies mitochondrial damage [31, 33, 44-46])
line 1024 The references are not specifically about mitochondria.
Response: We thank the reviewer for the important comment. As suggested, we have now incorporated the correct references (failure in neurons and glial cells [13, 21, 31, 44, 49]
lines 1029-1032 They absolutely must cite the seminal work by the J.D. Geiger lab.
Response: Thanks for your kind help in improving the manuscript, we have now rectified (cytochrome c and activation of intrinsic apoptotic pathways [29, 31, 44, 49, 50].
lines 1029-1034 The citations included (22. 33, 44, 45) are not appropriate.
Response: We thank the reviewer for the careful reading; we have now rectified and updated with appropriate references.
The entire lysosome section is seriously lacking and under-cited.
Response: We fully agree and have updated the section with more citations.
line 1110-1112 The citation (50) for Tat-induced structural changes is incorrect and again the seminal work of J.D. Geiger lab is warranted.
Response: Thanks for suggestion, we have now incorporated the Geiger lab references (CNS inflammation and neurodegeneration in people living with HIV [29-31, 44, 49].)
Note: In response to the reviewers’ suggestions, we have incorporated additional references at the appropriate locations throughout the manuscript, and the total number of citations (including newly added and reorganized references) is now 66.
Once again, we sincerely thank the reviewers for their careful evaluation and valuable recommendations, which have substantially improved the clarity, comprehensiveness, and impact of our manuscript.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors attempted to modify the manuscript according to suggestions by the reviewers, but their revisioned left me disappointed and concerned. I did not verify all sentences and references but there were many instances where sentences were left incited, citations included were not correct, and the most obvious and important citations were not included. Here are some examples.
Line 802 "Briefly," followed by multiple paragraphs! This is not correct use of the English language.
lines 995-996 They absolutely must cite the seminal literature by the J.D. Geiger lab showing the lysosome dysfunction precedes and amplifies mitochondrial damage. The following sentence has citations (22, 160, 179-184) that are at best questionable with regard to the content of the sentence.
line 1024 The references are not specifically about mitochondria.
lines 1029-1032 They absolutely must cite the seminal work by the J.D. Geiger lab.
lines 1029-1034 The citations included (22. 33, 44, 45) are not appropriate.
The entire lysosome section is seriously lacking and under-cited.
line 1110-1112 The citation (50) for Tat-induced structural changes is incorrect and again the seminal work of J.D. Geiger lab is warranted.
Author Response
Dear Editor,
We thank you for the opportunity to further revise our manuscript. We are pleased to submit the revised version in response to the reviewers’ comments. We sincerely appreciate the time and effort that you and the reviewers devoted to evaluating our work. The reviewers’ thoughtful and constructive feedback has significantly strengthened the manuscript.
In the revised version, we have carefully addressed all concerns raised by the reviewers and the editorial office. All revisions have been incorporated throughout the manuscript and are highlighted using track changes for ease of review.
We have attached both
1) Highlighted using track changes for ease of review.
2) Updated clean manuscript with all the changes accepted.
All authors have read and approved the revised manuscript. Should you have any questions or require additional information, please do not hesitate to contact us.
We have improved the language throughout the manuscript with particular attention to clarity, precision, and readability using a professional English-language editing service provided by our institution. However, to further ensure the highest linguistic and editorial standards and to maximize the impact and clarity of the work, we would greatly appreciate the assistance of the journal’s expert language editors, if needed.
Below, we provide a point-by-point response to each reviewer’s comment. We thank you again for your consideration and look forward to your response.
Sincerely,
Mudit Tyagi
On behalf of all authors
Response to the Reviewers:
We have improved the language throughout the manuscript with particular attention to clarity, precision, and readability using a professional English-language editing service provided by our institution. However, to further ensure the highest linguistic and editorial standards and to maximize the impact and clarity of the work, we would greatly appreciate the assistance of the journal’s expert language editors, if needed.
Line 802 "Briefly," followed by multiple paragraphs! This is not correct use of the English language.
Response: We appreciate the reviewer’s insightful suggestion. We have now revised the wording.
lines 995-996 They absolutely must cite the seminal literature by the J.D. Geiger lab showing the lysosome dysfunction precedes and amplifies mitochondrial damage. The following sentence has citations (22, 160, 179-184) that are at best questionable with regard to the content of the sentence.
Response: As suggested, we have now rectified. We have now included some more relevant references, including seminal references from Dr. J.D. Geiger lab (precedes and amplifies mitochondrial damage [31, 33, 44-46])
line 1024 The references are not specifically about mitochondria.
Response: We thank the reviewer for the important comment. As suggested, we have now incorporated the correct references (failure in neurons and glial cells [13, 21, 31, 44, 49]
lines 1029-1032 They absolutely must cite the seminal work by the J.D. Geiger lab.
Response: Thanks for your kind help in improving the manuscript, we have now rectified (cytochrome c and activation of intrinsic apoptotic pathways [29, 31, 44, 49, 50].
lines 1029-1034 The citations included (22. 33, 44, 45) are not appropriate.
Response: We thank the reviewer for the careful reading; we have now rectified and updated with appropriate references.
The entire lysosome section is seriously lacking and under-cited.
Response: We fully agree and have updated the section with more citations.
line 1110-1112 The citation (50) for Tat-induced structural changes is incorrect and again the seminal work of J.D. Geiger lab is warranted.
Response: Thanks for suggestion, we have now incorporated the Geiger lab references (CNS inflammation and neurodegeneration in people living with HIV [29-31, 44, 49].)
Note: In response to the reviewers’ suggestions, we have incorporated additional references at the appropriate locations throughout the manuscript, and the total number of citations (including newly added and reorganized references) is now 66.
Once again, we sincerely thank the reviewers for their careful evaluation and valuable recommendations, which have substantially improved the clarity, comprehensiveness, and impact of our manuscript.
Author Response File: Author Response.pdf
Round 3
Reviewer 1 Report
Comments and Suggestions for AuthorsCongratulations
Author Response
We sincerely thank you for your constructive feedback and guidance. Your insightful and intriguing suggestions have been highly valuable and have substantially improved the manuscript.
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThey continue to cite the wrong literature.
They continue to fail to cite statements that require the literature to be cited so readers can judge the accuracy of the statements.
They continue to fail to cite the seminal literature on which the statements are founded.
Author Response
They continue to cite the wrong literature.
Response: Unfortunately, we were unable to identify the incorrectly cited references. We would appreciate it if you could please specify which references you believe are cited incorrectly, so that we can address them precisely and make the necessary corrections.
They continue to fail to cite statements that require the literature to be cited so readers can judge the accuracy of the statements.
Response: We would appreciate it if you could please specify the exact statements that, in your opinion, require additional citations. This will allow us to address them accurately and ensure that the manuscript is fully compliant with the journal’s requirements.
They continue to fail to cite the seminal literature on which the statements are founded.
Response: We would be grateful if you could indicate the precise sections of the manuscript where seminal references are lacking. Your guidance will help us ensure that the manuscript appropriately reflects the foundational literature in the field.
Author Response File: Author Response.pdf
Round 4
Reviewer 2 Report
Comments and Suggestions for AuthorsHere are examples of problems with the citations.
lines 82-83. I do not think citations 7 and 18 are appropriate.
lines 698-699. Citations 144 and 147 are not appropriate.
Section 6 (lines 802-821) has no citations for sections on NNRTIs, NRTIs, PIs, INSTIs and entry inhibitors.
line 1044. Citations 95 and 96 are not appropriate.
The following manuscripts are foundational to what they are writing about.
Hui, L., Ye, Y., Soliman, M.L., Lakpa, K.L., Miller, N.M., Afghah, A., Geiger, J.D. and X. Chen (2019) Antiretroviral drugs promote amyloidogenesis by de-acidifying endolysosomes. J. Neuroimmune Pharmacology 2019 Jul 23. doi: 10.1007/s11481-019-09862-1.
Khan, N., Haughey, N.J., Nath, A. and J.D. Geiger (2019) Involvement of organelles and inter-organellar signaling in the pathogenesis of HIV-1 neurocognitive disorder and Alzheimer’s disease. Brain Research 2019 Nov 1;1722:146389. doi: 10.1016/j.brainres.2019.146389
Afghah, Z., Chen X. and J.D. Geiger. (2020) Role of endolysosomes and inter-organellar signaling in brain disease. Neurobiology of Disease Feb;134:104670. doi:10.1016/j.nbd.2019.104670
Lakpa K.L., Khan N., Afghah Z., Chen X. and J.D. Geiger (2021) Lysosomal Stress Response(LSR): Physiological Importance and Pathological Relevance, (Invited review) J. Neuroimmune Pharmacol. 16(2): 219-237 https://doi.org/10.1007/s11481-021-09990-7
Halcrow, P., Lakpa, K.L., Khan, N., Chen, X. and J.D. Geiger. (2022) HIV-1 gp120-Induced Endolysosome De-Acidification Leads to Efflux of Endolysosome Iron, and Increases in Mitochondrial Iron and Reactive Oxygen Species. J. Neuroimune Pharmacol. 17, 181-194 PMID: 33834418
Halcrow, P.W., Kumar, N., Quansah, D.N.K., Liang, B. and J.D. Geiger (2022) Endolysosome Iron Chelation Inhibits HIV-1 Protein-Induced Endolysosome Deacidification-Induced Increases in Mitochondrial Fragmentation, Mitophagy, and Cell Death. Cells. 2022 May 31;11(11):1811. doi: 10.3390/cells11111811. PMID: 35681506
Halcrow, P.W., Kumar, N., Hao, E., Khan, N., Meucci, O. and J.D. Geiger. (2023) Mu Opioid Receptor-Mediated Release of Endolysosome Iron Increases Levels of Mitochondrial Iron, Reactive oxygen Spoecies, and Cell Death. Journal of NeuroImmune Pharmacology and Therapeutics, 25: 19-35. doi: 10.1007/s11481-021-09995-2.
Halcrow, P.W., Quansah, D.N.K., Kumar, N., Solloway, R.L., Teigen, K.M., Lee, K.A., Liang, and J.D. Geiger. (2024) Weak Base Drug-Induced Endolysosome Iron Dyshomeostasis Controls the Generation of Reactive Oxygen Species, Mitochondrial Depolarization, and Cytotoxicity. Journal of NeuroImmune Pharmacology and Therapeutics Neuroimmune Pharmacol. Ther. http://doi.org/10.1515/nipt-2023-0021.
Kumar, N., Halcrow, P.W., Quansah, D.N.K., Liang, B., Meucci, O. and J.D. Geiger (2025) Involvement of Endolysosome Iron in HIV-1 gp120-, Morphine-, and Iron Supplementation Induced Disruption of the Reactive Species Interactome and Induction of Neurotoxicity. Redox Report DOI: 10.1080/13510002.2025.2546496
Comments on the Quality of English LanguageN/A
Author Response
lines 82-83. I do not think citations 7 and 18 are appropriate.
Response: We have incorporated the suggested references. Reference 7 discusses the role of cytokines in both HIV latency and neurotoxicity, whereas Reference 18 addresses HIV-induced pathogenesis, including neuropathies and neurotoxic effects. In addition, we have included several further citations to strengthen the supporting literature.
It should also be noted that this sentence serves only as an introductory statement, with a more comprehensive and detailed discussion of this topic provided in the subsequent section.
The neurotoxic effects of HIV are increasingly recognized as both direct and indirect, contributing to neurodegeneration [7, 18].
- Hokello, J.; Sharma, A. L.; Dimri, M.; Tyagi, M., Insights into the HIV Latency and the Role of Cytokines. Pathogens 2019, 8, (3).
- Sonti, S.; Sharma, A. L.; Tyagi, M., HIV-1 persistence in the CNS: Mechanisms of latency, pathogenesis and an update on eradication strategies. Virus Res 2021, 303, 198523.
lines 698-699. Citations 144 and 147 are not appropriate.
Response: We appreciate the reviewer’s comment. References 144 and 147 are appropriate in this context, as they specifically address the effects of HIV and its proteins, particularly Tat and gp120, on blood–brain barrier (BBB) disruption and neurotoxicity. Nevertheless, we have now added additional relevant references to further strengthen the supporting literature.
Accordingly, the sentence has been retained and clarified as follows:
Viral proteins, primarily gp120 and Tat, directly impair BBB integrity by upregulating matrix metalloproteinases (MMP-2 and MMP-9) and promoting degradation of key tight-junction proteins, including claudin-5, occludin, and ZO-1 [144,147].
- Said, N.; Venketaraman, V., Neuroinflammation, Blood-Brain Barrier, and HIV Reservoirs in the CNS: An In-Depth Exploration of Latency Mechanisms and Emerging Therapeutic Strategies. Viruses 2025, 17, (4).
- Jadhav, S.; Nema, V., HIV-Associated Neurotoxicity: The Interplay of Host and Viral Proteins. Mediators Inflamm 2021, 2021, 1267041.
Section 6 (lines 802-821) has no citations for sections on NNRTIs, NRTIs, PIs, INSTIs and entry inhibitors.
Response: While the section titled “ART-associated neurotoxicity” already included relevant supporting references, we have incorporated additional citations as suggested to further strengthen and broaden the literature coverage.
line 1044. Citations 95 and 96 are not appropriate.
Response: Although the previously cited references addressed the relevant concept, we have now added and replaced them with more specific and directly applicable citations, including those recommended in the subsequent comment, to improve precision and clarity of the supporting literature.
Similarly, several antiretroviral agents, particularly older NRTIs and some protease inhibitors, disrupt lysosomal membrane integrity, alter intra-lysosomal pH, and inhibit cathepsin activity, resulting in accumulation of dysfunctional organelles and undegraded protein aggregates [95, 96].
- Lewis, W.; Day, B. J.; Copeland, W. C., Mitochondrial toxicity of NRTI antiviral drugs: an integrated cellular perspective. Nat Rev Drug Discov 2003, 2, (10), 812-22.
- Apostolova, N.; Blas-Garcia, A.; Esplugues, J. V., Mitochondrial toxicity in HAART: an overview of in vitro evidence. Curr Pharm Des 2011, 17, (20), 2130-44.
The following manuscripts are foundational to what they are writing about.
Hui, L., Ye, Y., Soliman, M.L., Lakpa, K.L., Miller, N.M., Afghah, A., Geiger, J.D. and X. Chen (2019) Antiretroviral drugs promote amyloidogenesis by de-acidifying endolysosomes. J. Neuroimmune Pharmacology 2019 Jul 23. doi: 10.1007/s11481-019-09862-1.
Khan, N., Haughey, N.J., Nath, A. and J.D. Geiger (2019) Involvement of organelles and inter-organellar signaling in the pathogenesis of HIV-1 neurocognitive disorder and Alzheimer’s disease. Brain Research 2019 Nov 1;1722:146389. doi: 10.1016/j.brainres.2019.146389
Afghah, Z., Chen X. and J.D. Geiger. (2020) Role of endolysosomes and inter-organellar signaling in brain disease. Neurobiology of Disease Feb;134:104670. doi:10.1016/j.nbd.2019.104670
Lakpa K.L., Khan N., Afghah Z., Chen X. and J.D. Geiger (2021) Lysosomal Stress Response(LSR): Physiological Importance and Pathological Relevance, (Invited review) J. Neuroimmune Pharmacol. 16(2): 219-237 https://doi.org/10.1007/s11481-021-09990-7
Halcrow, P., Lakpa, K.L., Khan, N., Chen, X. and J.D. Geiger. (2022) HIV-1 gp120-Induced Endolysosome De-Acidification Leads to Efflux of Endolysosome Iron, and Increases in Mitochondrial Iron and Reactive Oxygen Species. J. Neuroimune Pharmacol. 17, 181-194 PMID: 33834418
Halcrow, P.W., Kumar, N., Quansah, D.N.K., Liang, B. and J.D. Geiger (2022) Endolysosome Iron Chelation Inhibits HIV-1 Protein-Induced Endolysosome Deacidification-Induced Increases in Mitochondrial Fragmentation, Mitophagy, and Cell Death. Cells. 2022 May 31;11(11):1811. doi: 10.3390/cells11111811. PMID: 35681506
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Response: We sincerely appreciate the reviewer’s effort and helpful suggestions. The recommended citations have been incorporated alongside other key references already included in the manuscript to further strengthen the supporting literature.
Author Response File: Author Response.pdf
