Brain-Homing Peptide Expression on the Membrane Enhances the Delivery of Exosomes to Neural Cells and Tissue
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsAfter review the manuscript Brain-homing peptaide expresion...
I consider that it is clear at the present form.
It is a presentation of a mechanism to deliver peptides and other molecules in the possibility to cross the BBB.
Author Response
Thank you for taking the time to review our manuscript, "Brain-homing peptide expression...". We sincerely appreciate your thoughtful evaluation and are pleased to hear that you find the manuscript clear and the content meaningful in presenting a mechanism to deliver peptides and molecules with the potential to cross the blood-brain barrier (BBB).
Your positive feedback affirms the direction of our research and motivates us to continue exploring this important area. We are grateful for your time and effort in reviewing our work.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe topic at hand seems relevant.
The idea of using exosomes rather than liposomes is intriguing. I am not sufficiently introduced in the field to know how novel this idea is; I think the Authors should spend some words on this, citing possible papers in which this idea of having cells express the targeting peptide on exosomes rather than enriching a liposome with the same peptide, and in general argumenting why they think that this approach may yield interesting results, as opposed to the idea of targeted liposomes, which had appeared as the revolution in tharapy, some 20 years back, but were actually successful in only a few cases.
Also, I am not able to judge the potential of this approach in being translated in the clinical arena; again, some notes on this possibility and the related difficulties and concerns would add value to the discussion.
Overall, I think that the experimental aspects are sound, and the paper may deserve to be published if the Authors are able to stress the novelty and originality of the approach (i.e. not many other papers have attempted at producing targeted exosomes), its safety (the cells have to be transfected using a viral vector, which is not supposed to leave any track on the exosomes, but this aspect should be specifically addressed), and its feasibility.
Author Response
Comment: The topic at hand seems relevant.
The idea of using exosomes rather than liposomes is intriguing. I am not sufficiently introduced in the field to know how novel this idea is; I think the Authors should spend some words on this, citing possible papers in which this idea of having cells express the targeting peptide on exosomes rather than enriching a liposome with the same peptide, and in general argumenting why they think that this approach may yield interesting results, as opposed to the idea of targeted liposomes, which had appeared as the revolution in tharapy, some 20 years back, but were actually successful in only a few cases.
Also, I am not able to judge the potential of this approach in being translated in the clinical arena; again, some notes on this possibility and the related difficulties and concerns would add value to the discussion.
Overall, I think that the experimental aspects are sound, and the paper may deserve to be published if the Authors are able to stress the novelty and originality of the approach (i.e. not many other papers have attempted at producing targeted exosomes), its safety (the cells have to be transfected using a viral vector, which is not supposed to leave any track on the exosomes, but this aspect should be specifically addressed), and its feasibility.
Response to Reviewer:
We thank the reviewer for their insightful comment. In response, we have expanded the discussion section to emphasize the novelty and originality of our approach. Specifically, we have included additional references (e.g., Alvarez-Erviti et al., Kamerkar et al.) to highlight prior studies on targeted exosomes and provide context for our work. Additionally, we have critically compared exosomes with liposomes, discussing their inherent advantages in biocompatibility, targeting capabilities, and clinical potential. To address safety concerns, we have incorporated a detailed assessment of challenges, including the absence of residual viral vectors in exosome preparations. These revisions, detailed in Lines 299–327, aim to underscore the feasibility and translational relevance of our work.
Reviewer 3 Report
Comments and Suggestions for AuthorsThis manuscript is devoted to brain-homing peptide expression. It was shown that it enhances the delivery of exosomes to neural cells and tissue. These nano vesicles can cross cellular boundaries, making them ideal vehicles for targeted drug delivery to the brain. The findings of this work may be applied for innovative treatments that could significantly enhance patients' quality of life. I think that this manuscript may be published after minor revision.
Notes:
1. In the Figure 2B the scale values should be changed and presented in a more readable form. Also the scale values and labels in the Figure 3A, B should be increased for clarity.
2. Authors reported that the majority of the exosomes remained within the expected 30–100 nm range, even after PEG processing. But they nothing mention about the dispersity of these exosomes. Monodisperse or polydisperse systems are formed? Short comment should be added.
3. What is the advantage of the developed in this work exosomes comparing with the known in the literature? Short comment should be added in Conclusions.
Author Response
Comment: This manuscript is devoted to brain-homing peptide expression. It was shown that it enhances the delivery of exosomes to neural cells and tissue. These nano vesicles can cross cellular boundaries, making them ideal vehicles for targeted drug delivery to the brain. The findings of this work may be applied for innovative treatments that could significantly enhance patients' quality of life. I think that this manuscript may be published after minor revision.
Response to the comment: We sincerely thank the reviewer for their positive evaluation of our manuscript and for acknowledging the significance of our findings. We are pleased that the potential applications of brain-homing peptides for targeted drug delivery and their implications for improving patients' quality of life resonate with you. We have carefully addressed the minor revision suggestions (detailed in subsequent responses), ensuring the manuscript meets the highest standards of clarity and scientific rigor. Your feedback has been invaluable in refining the presentation of our work, and we look forward to its publication.
Comment 1: In the Figure 2B the scale values should be changed and presented in a more readable form. Also the scale values and labels in the Figure 3A, B should be increased for clarity.
Response to the comment1: Thank you for highlighting this issue. We have updated Figure 2B to present the scale values in a more legible format. Additionally, we have enlarged the scale values and labels in Figures 3A and 3B to enhance their clarity. The revised figures have been incorporated into the updated manuscript for your review.
Comment 2: Authors reported that the majority of the exosomes remained within the expected 30–100 nm range, even after PEG processing. But they nothing mention about the dispersity of these exosomes. Monodisperse or polydisperse systems are formed? Short comment should be added.
Response to the comment 2: Thank you for suggesting the inclusion of data on the dispersity of the exosomes. We have conducted an analysis using dynamic light scattering (DLS), which confirmed that the exosomes exhibit a monodisperse distribution with a low polydispersity index (PDI). A discussion of the dispersity has been added to the Results section of the revised manuscript (Lines 236–238).
Comment 3: What is the advantage of the developed in this work exosomes comparing with the known in the literature? Short comment should be added in Conclusions.
Responce to the comment 3: Thank you for bringing up this important point. In response, we have added a concise statement in the Conclusions section to emphasize the key advantages of the exosomes developed in this study. Specifically, we underscore their enhanced targeting efficiency facilitated by brain-homing peptide expression, their improved ability to cross the blood-brain barrier, and their maintained size and stability following PEG processing. These attributes distinguish our exosomes from others reported in the literature and highlight their potential for clinical applications. (Lines 370–374)
