Auranofin as a Novel Anticancer Drug for Anaplastic Thyroid Cancer
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsI carefully read the manuscript entitled “Auranofin as a novel anticancer drug for anaplastic thyroid cancer".
To improve it, some comments must be made.
The comments are as follows:
- It is suggested that the keywords be organized based on the PubMed MeSH terms and rearranged according to the English alphabet.
- It is suggested to imply the epidemiology of anaplastic thyroid cancer in the “introduction” section.
- It is suggested that common treatments for anaplastic thyroid cancer be included in the “introduction” section.
- Please clarify the aim(s) of the present study at the end of the "Introduction" section.
- Some of the methods need to use relevant reference(s).
- Please clarify the kind of qPCR. (Reverse transcription PCR is used to make cDNA and is not a quantitative PCR.) I think the author means real-time PCR.
- The primers (E-cad, ITGB1, and BAX) must be checked.
- It is mandatory to change qualitative data to quantitative and analyze part C of Figures 3, and 4, part B of Figure 6, and part F of Figure 7.
- The manuscript needs some minor English editing.
Best regards
Author Response
MDPI pharmaceuticals
# 3261539
Title: Auranofin as a novel anticancer drug for anaplastic thyroid cancer
Dear Editors-in-Chief:
We are grateful to you and the reviewers for putting time and effort into reviewing our manuscript. The detailed and constructive comments provided valuable insights into our study. We are submitting the revised manuscript and highlighted the changes made to the text in blue color in response to your suggestions.
<Reviewer 1>
General comment:
I carefully read the manuscript entitled “Auranofin as a novel anticancer drug for anaplastic thyroid cancer".
[Comment 1]
|
It is suggested that the keywords be organized based on the PubMed MeSH terms and rearranged according to the English alphabet. |
Response: We would like to express our sincere appreciation for your thoughtful and constructive comments on our manuscript. Your detailed review has provided valuable insights that greatly contributed to improving the quality of our study.
Thank you for your valuable suggestions. In accordance with your recommendation, we have revised the manuscript and reordered the keywords based on the PubMed MeSH terms in alphabetical order. The updated keywords have been inserted into the manuscript accordingly.
[Comment 2]
|
It is suggested to imply the epidemiology of anaplastic thyroid cancer in the “introduction” section. |
Response: Thank you for your valuable feedback. In accordance with your suggestion, we have added the epidemiology of anaplastic thyroid cancer to the introduction section of the manuscript. We appreciate your insights and hope the revisions meet your expectations.
[Comment 3]
|
It is suggested that common treatments for anaplastic thyroid cancer be included in the “introduction” section. |
Response: Thank you for your comments. As per your suggestion, we have added information about common treatments for anaplastic thyroid cancer in the introduction section.
[Comment 4]
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Please clarify the aim(s) of the present study at the end of the "Introduction" section. |
Response: In accordance with the reviewer's suggestion, we have clarified the aim of this study more explicitly at the end of the "Introduction" section. We greatly appreciate your valuable input.
[Comment 5]
|
Some of the methods need to use relevant reference(s). |
Response: Thank you for your valuable feedback. In response to your suggestions, we have added the appropriate references to the respective sections detailing the mRNA-seq and RT-PCR methods in the manuscript.
[Comment 6]
|
Please clarify the kind of qPCR. (Reverse transcription PCR is used to make cDNA and is not a quantitative PCR.) I think the author means real-time PCR. |
Response: We would like to clarify that the method we employed is Real-Time Quantitative Reverse Transcription PCR (Real-Time qRT-PCR), which corresponds to the Real-Time PCR mentioned by the reviewer. To avoid any confusion, we have revised the terminology in accordance with your suggestion. We sincerely appreciate your thoughtful feedback.
[Comment 7]
|
The primers (E-cad, ITGB1, and BAX) must be checked. |
Response: We have rechecked the primers for E-cad, ITGB1, and BAX in response to your comment. We sincerely appreciate your valuable feedback.
[Comment 8]
|
It is mandatory to change qualitative data to quantitative and analyze part C of Figures 3, and 4, part B of Figure 6, and part F of Figure 7. |
Response: We sincerely appreciate your invaluable comment. In response, we have converted the qualitative data to quantitative and updated the graphs accordingly for part C of Figures 3 and 4, part B of Figure 6, and part F of Figure 7. Your thoughtful feedback has been instrumental in refining our work, and we are truly grateful for it.
[Comment 9]
|
The manuscript needs some minor English editing. |
Response: We have completed the English editing of the manuscript, and the certificate has been submitted as an additional file. Should further revisions be required, we are prepared to utilize MDPI's editing services. We sincerely appreciate your thorough review.
Corresponding Author
Soonchul Lee, M.D., Ph.D.
Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University
59 Yatap-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea
Zip code: 13496
Tel. +82-31-780-5289, Fax.+ 82-31-708-3578
E-mail address: [email protected]
Hyun-Ju An, Ph.D.
Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University
59 Yatap-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea
Zip code: 13496
Tel. +82-31-780-5289, Fax.+ 82-31-708-3578
E-mail address: [email protected]
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript reported repositioning of auranofin for anaplastic thyroid cancer (ATC) by in vitro and in vivo biological evaluation Antitumor efficacy was also evaluated through gene and protein expression analysis. Key molecular pathways were identified, including those involved in apoptosis and reactive oxygen species (ROS) production. In vivo experiments, confirmed auranofin’s anticancer effects.
I have some question that should be addressed:
Auranofin is in clinical trials for several different types of blood cancer cells. This was not reported in the introduction section. In addition, the direct inhibition of TrxR was not the only target of Auranofin. As additiona direct targets, proteasome associated deubiquitinases, exokinase, PKCiota and IkB kinase shoul be cited
The authors have reported that this compound increase reactive oxygen species (ROS) production, and this mechanism should be investigated evaluating the potential inhibition thioredoxin reductase (TrxR). Inhibiting TrxR dysregulates the intracellular redox state causing increased intracellular reactive oxygen species levels.
Can the authors evaluate the expression level of TrxR in 8505C and FRO cells to support its potential involvement in ROS production?
In vivo studies were performed at very high doses of Auronafin (100 micromolar) and without a positive control. How is the effect at lower doses? In addition, Auranofin was injected intratumorally. It is an orally administered drug and is considered safe for humans. Why the authors have not considered the intragastric administration at different doses?
Author Response
MDPI pharmaceuticals
# 3261539
Title: Auranofin as a novel anticancer drug for anaplastic thyroid cancer
Dear Editors-in-Chief:
We are grateful to you and the reviewers for putting time and effort into reviewing our manuscript. The detailed and constructive comments provided valuable insights into our study. We are submitting the revised manuscript and highlighted the changes made to the text in blue color in response to your suggestions.
<Reviewer 2>
General comment:
The manuscript reported repositioning of auranofin for anaplastic thyroid cancer (ATC) by in vitro and in vivo biological evaluation Antitumor efficacy was also evaluated through gene and protein expression analysis. Key molecular pathways were identified, including those involved in apoptosis and reactive oxygen species (ROS) production. In vivo experiments, confirmed auranofin’s anticancer effects.
[Comment 1]
|
Auranofin is in clinical trials for several different types of blood cancer cells. This was not reported in the introduction section |
Response: We would like to express our sincere appreciation for your thoughtful and constructive comments on our manuscript. Your detailed review has provided valuable insights that greatly contributed to improving the quality of our study.
As per your suggestion, we have included information in the introduction regarding the ongoing clinical trials of auranofin for several different types of blood cancer.
[Comment 2]
|
In addition, the direct inhibition of TrxR was not the only target of Auranofin. As additiona direct targets, proteasome associated deubiquitinases, exokinase, PKCiota and IkB kinase shoul be cited |
Response: Thank you for your insightful and constructive comments. In response to your suggestions, we have expanded the content in both the Introduction and Discussion sections. Your feedback has significantly enriched the depth and quality of our manuscript, and we sincerely appreciate your valuable input.
[Comment 3]
|
The authors have reported that this compound increase reactive oxygen species (ROS) production, and this mechanism should be investigated evaluating the potential inhibition thioredoxin reductase (TrxR). Inhibiting TrxR dysregulates the intracellular redox state causing increased intracellular reactive oxygen species levels. Can the authors evaluate the expression level of TrxR in 8505C and FRO cells to support its potential involvement in ROS production? |
Response: Upon reviewing several studies, we recognize that while auranofin was originally developed as a treatment for rheumatoid arthritis, it has exhibited anticancer efficacy in animal models and has been approved for clinical trials in lung and ovarian cancers. As the reviewer aptly noted, auranofin’s most recognized target is Thioredoxin reductase, with its primary mechanism involving the elevation of ROS levels, leading to cell death. However, in our study, it remains unclear whether the key pathway in thyroid cancer cell lines is directly linked to Thioredoxin reductase. To investigate the main pathways regulated in ATC, which may differ from other cancers, we conducted mRNA-sequencing and have made all results publicly available via the GEO database. Our analysis of the raw data revealed minor variations in TrxR types among the cell lines. We propose that in thyroid cancer, there may be additional critical targets beyond Thioredoxin reductase. Therefore, in this study, we focused on a comprehensive gene expression analysis using KEGG and GOBP pathways. While Thioredoxin reductase is indeed a significant mechanism, as pointed out by the reviewer, we will continue to explore the co-regulated mechanisms in future studies. We sincerely appreciate the reviewer's insightful and detailed feedback.
- Zhang X, Selvaraju K, Saei AA, D'Arcy P, Zubarev RA, Arnér ES, Linder S. Repurposing of auranofin: Thioredoxin reductase remains a primary target of the drug. Biochimie. 2019 Jul;162:46-54
- Johnson SS, Liu D, Ewald JT, Robles-Planells C, Christensen KA, Bayanbold K, Wels BR, Solst SR, O'Dorisio MS, Allen BG, Menda Y, Spitz DR, Fath MA. Auranofin Inhibition of Thioredoxin Reductase Sensitizes Lung Neuroendocrine Tumor Cells (NETs) and Small Cell Lung Cancer (SCLC) Cells to Sorafenib as well as Inhibiting SCLC Xenograft Growth. bioRxiv [Preprint]. 2024 Jan 30:2023.05.07.539772. doi: 10.1101/2023.05.07.539772. Update in: Cancer Biol Ther. 2024 Dec 31;25(1):2382524.
[Comment 4]
|
In vivo studies were performed at very high doses of Auronafin (100 micromolar) and without a positive control. How is the effect at lower doses? |
Response: -
[Comment 5]
|
In addition, Auranofin was injected intratumorally. It is an orally administered drug and is considered safe for humans. Why the authors have not considered the intragastric administration at different doses? |
Response: Subcutaneous (SC) injection offers several advantages over intragastric administration in mouse tumor experiments. It provides higher bioavailability by bypassing the digestive system, leading to more consistent drug absorption. SC injections also allow for more precise dosing, ensuring accurate drug delivery to the bloodstream, avoiding the variability seen with oral administration. Additionally, SC injection is advantageous for drugs with poor gastrointestinal absorption, enabling a larger portion of the drug to reach circulation. Furthermore, localized drug delivery through SC administration can enhance therapeutic effects and reduce systemic toxicity, making it a preferred method in tumor models where consistent and effective drug delivery is crucial.
We fully agree with the reviewer's perspective. However, we believe that, given the advantages of subcutaneous (SC) injection outlined above, it is more suitable for our experimental conditions, particularly considering the challenges we face in fine-tuning the dose of auranofin via intragastric administration. SC injection offers a more consistent and controlled approach, which is crucial in our study. That being said, we acknowledge that the limitations raised by the reviewer are valid and represent an important aspect of our work. We will therefore address this point by including it as a limitation in the Discussion section. We sincerely appreciate the reviewer's constructive feedback, which has helped to strengthen our manuscript.
Corresponding Author
Soonchul Lee, M.D., Ph.D.
Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University
59 Yatap-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea
Zip code: 13496
Tel. +82-31-780-5289, Fax.+ 82-31-708-3578
E-mail address: [email protected]
Hyun-Ju An, Ph.D.
Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University
59 Yatap-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea
Zip code: 13496
Tel. +82-31-780-5289, Fax.+ 82-31-708-3578
E-mail address: [email protected]
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsDear authors thank you for your kind responses.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors have addressed to all my comments. I suggest to publish the manuscript in the present form.
