Targeting the Hippo- Yes-Associated Protein/Transcriptional Coactivator with PDZ-Binding Motif Signaling Pathway in Primary Liver Cancer Therapy

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Editor,

The manuscript “Targeting the Hippo-YAP/TAZ Signaling Pathway in Primary Liver Cancer Therapy” by Yina Wang and Liangyou Rui, has focused on various aspect of Hippo pathway potentially suitable for drug design. The authors discussed the main elements of the pathway, the regulators, potential targets and candidate inhibitors. The manuscript seems to be interested for Onco audiences. However, before being considered for publication, the text should be carefully reviewed for structure as well as several ambiguities. I would suggest the most significant items as below:

1-      Introduction can be more concise by reducing the repetitions.

2-      A schematic pathway that shows the main factors in signaling and the targeting points would help a lot.

3-      The incidence rate of liver cancer (Line 39) is better to be checked and updated.

4-      Several cases of ambiguous, contradictory or incorrect sentences are seen including lines 116, 119, 190, 200, 242, 247, … .

5-      All of the plant or animal names must be in italic form.

6-      Also it needs a careful revision for the grammatical fault or typos; i.e. the short names such as LATS1/2 has appeared incorrect frequently.

Best regards,

Comments on the Quality of English Language

Language revision of the manuscript is very necessary and important. 

Author Response

Thank you very much for taking the time and effort to review our manuscript. We appreciate your positive and constructive feedback, and we truly believe that during the revision we improved our manuscript to address your comments and concerns.

Please find our point-to-point response below, and any changes or revisions to the article can be highlighted (marked as red color) in the revised form of the manuscript. We remain at your disposition for further clarifications and improvements.

Comments 1-      Introduction can be more concise by reducing the repetitions.

Response 1: Thank you for pointing this out. We agree with this comment. Therefore, we have reduced the repetitions in Introduction and Part 2.

Comments 2-      A schematic pathway that shows the main factors in signaling and the targeting points would help a lot.

Response 2: Agree. We have, accordingly, added Figure 1 to emphasize this point. Please see Figure 1 in the revise manuscript.

Comments 3-      The incidence rate of liver cancer (Line 39) is better to be checked and updated.

Response 3: Thank you so much for your comment. The latest global incidence rate of liver cancer that can be found in the existing literature is from 2020, and we hope that an updated report will be produced soon.

Comments 4-      Several cases of ambiguous, contradictory or incorrect sentences are seen including lines 116, 119, 190, 200, 242, 247, … .

Response 4 : Thanks a lot for pointing this out. We have corrected and checked through the text to avoid such mistakes.

Comments 5-      All of the plant or animal names must be in italic form.

Response 5: Agree. We have, accordingly, revised the plant or animal names into italic form.

Comments 6-      Also it needs a careful revision for the grammatical fault or typos; i.e. the short names such as LATS1/2 has appeared incorrect frequently.

Response 6: Thank you for pointing this out. We agree with this comment. Therefore, we have checked through the text to avoid grammatical fault or typos.

Reviewer 2 Report

Comments and Suggestions for Authors

This review provides a comprehensive summary of recent advances in the Hippo-YAP/TAZ pathway in primary liver cancer and highlights its therapeutic potential for patients. It is good. Some points should be noted as below,

1) More advances should be added, for example, in line 161, “3.3 Tumor invasion and metastasis”:

①Guan L, Li T, Ai N, Wang W, He B, Bai Y, Yu Z, Li M, Dong S, Zhu Q, Ding XX, Zhang S, Li M, Tang G, Xia X, Zhao J, Lin S, Yao S, Zhang L, Chen G, Liu FE, Li X, Zhang H.MEIS2C and MEIS2D promote tumor progression via Wnt/beta-catenin and hippo/YAP signaling in hepatocellular carcinoma.J Exp Clin Cancer Res. 2019 Oct 17;38(1):417.

③Yan YC, Meng GX, Yang CC, Yang YF, Tan SY, Yan LJ, Ding ZN, Ma YL, Dong ZR, Li T.Diacylglycerol lipase alpha promotes hepatocellular carcinoma progression and induces lenvatinib resistance by enhancing YAP activity.Cell Death Dis. 2023 Jul 6;14(7):404.

2) This review primarily provides a molecular-level description. However, it is important to acknowledge that cancer is a complex pathological ecosystem, as recently described in an article (https://pubmed.ncbi.nlm.nih.gov/37056571/). This paper is suggested to be reviewed. It should include some discussion at the end of this paper to indicate that merely approaching cancer from a molecular perspective may not address all cancer-related issues.

 

 

 

Author Response

Thank you very much for taking the time and effort to review our manuscript. We appreciate your positive and constructive feedback, and we truly believe that during the revision we improved our manuscript to address your comments and concerns.

Please find our point-to-point response below, and any changes or revisions to the article can be highlighted (marked as red color) in the revised form of the manuscript. We remain at your disposition for further clarifications and improvements.

This review provides a comprehensive summary of recent advances in the Hippo-YAP/TAZ pathway in primary liver cancer and highlights its therapeutic potential for patients. It is good. Some points should be noted as below,

Comments 1) More advances should be added, for example, in line 161, “3.3 Tumor invasion and metastasis”:

①Guan L, Li T, Ai N, Wang W, He B, Bai Y, Yu Z, Li M, Dong S, Zhu Q, Ding XX, Zhang S, Li M, Tang G, Xia X, Zhao J, Lin S, Yao S, Zhang L, Chen G, Liu FE, Li X, Zhang H.MEIS2C and MEIS2D promote tumor progression via Wnt/beta-catenin and hippo/YAP signaling in hepatocellular carcinoma.J Exp Clin Cancer Res. 2019 Oct 17;38(1):417.

③Yan YC, Meng GX, Yang CC, Yang YF, Tan SY, Yan LJ, Ding ZN, Ma YL, Dong ZR, Li T.Diacylglycerol lipase alpha promotes hepatocellular carcinoma progression and induces lenvatinib resistance by enhancing YAP activity.Cell Death Dis. 2023 Jul 6;14(7):404.

Response 1: Thank you for pointing this out. We agree with this comment. Therefore, we have added these references, please see ref 50-51 and Line 169-176.

Comments 2) This review primarily provides a molecular-level description. However, it is important to acknowledge that cancer is a complex pathological ecosystem, as recently described in an article (https://pubmed.ncbi.nlm.nih.gov/37056571/). This paper is suggested to be reviewed. It should include some discussion at the end of this paper to indicate that merely approaching cancer from a molecular perspective may not address all cancer-related issues.

Response 2: Thanks a lot for your comment. We agree with this comment. Therefore, we have added discussion on this topic, please see ref 98 and Line 334-342.

Reviewer 3 Report

Comments and Suggestions for Authors

The current manuscript is reviewing the Hippo signalling pathway as a possible therapeutic target in liver cancer. This is important review as liver cancers have poor prognosis and low survival rates. The manuscript covered the most important aspects of this pathway and is generally well-written.  However, the Authors are asked to address the following issues:

1.      Line 24, what is meant by Hippo/YAP/TAZZ pathway? The double “ZZ” seems to be odd?

2.      Line 58,  MST1/2 phosphorylate activate or MST1/2 phosphorylate and activate?

3.      Line 60, YAP/TAZ and remains YAP/TAZ in the cytoplasm. I am not sure that “remain” is the correct work, please consider re-wording (e.g. retain).  

4.      Line 78, Please consider adding a figure to show this signalling pathway as this will help the readers to understand the signalling pathway.

5.      Line 119, the Authors mention that YAP is methylated, and this increases its oncogenicity by preventing it nuclear translocation. The Authors are asked to explain this further as they mentioned in lines 84 and 85 that YAP nuclear translocation activate target genes and target genes are mainly proliferative and survival genes (i.e. oncogenic). Then preventing YAP nuclear translocation should reduce oncogenicity not increasing. Please clarify.

6.      Lines 154 to 160, It is not clear for me as the Authors first indicate that impaired autophagy increases YAP but they conclude in the last non-referenced statement (lines 158 to 160), that YAP suppresses autophagy. This seems like vicious circle. Would the Authors be able to clarify this further? And add reference to the final statement (lines 158 to 160)?

7.      Lines 164 to 167, The Authors describe the effects of TAZ knockdown in liver cancer cells, but these effects do not seem to align with the effects of TAZ knockout which causes YAP overexpression as described in lines 89 to 90. Would the Authors be able to clarify this point and whether knockdown and knockout have different effects?

8.      Lines 190 to 192, the effect of hypoxia seems to be inducing YAP and its relocation to nucleus, so it is not clear for me how this would increase the sensitivity of HCC cells to irinotecan? Would the Authors be able to clarify?

9.      Lines 272 to 274, The authors describe that the effects of poplar propolis through LAST1 dependent manner but in table 3, they indicate that it is thorough LAST2, please clarify.

Lines 299 to 305, the Authors describe targeting the Hippo/YAP/TAZ pathway by single agent medications. Are there any studies which targeted this pathway by combination therapy? Drug combinations are very common in cancer treatment and because Hippo/YAP/TAZ pathway involves several proteins and interacts with other pathways (e.g. MAPK), then it is sensible to target the pathway with combination therapy. The Authors are invited to comment.   

Author Response

Thank you very much for taking the time and effort to review our manuscript. We appreciate your positive and constructive feedback, and we truly believe that during the revision we improved our manuscript to address your comments and concerns.

Please find our point-to-point response below, and any changes or revisions to the article can be highlighted (marked as red color) in the revised form of the manuscript. We remain at your disposition for further clarifications and improvements.

The current manuscript is reviewing the Hippo signalling pathway as a possible therapeutic target in liver cancer. This is important review as liver cancers have poor prognosis and low survival rates. The manuscript covered the most important aspects of this pathway and is generally well-written.  However, the Authors are asked to address the following issues:

Comments 1.      Line 24, what is meant by Hippo/YAP/TAZZ pathway? The double “ZZ” seems to be odd?

Response 1: Thank you for pointing this out. We have revised this typo into correct “Hippo-YAP/TAZ pathway”.

Comments 2.      Line 58,  MST1/2 phosphorylate activate or MST1/2 phosphorylate and activate?

Response 2: Thank you for pointing this out. We have revised this typo into “MST1/2 phosphorylate and activate LATS1/2, and LATS1/2 in turn phosphorylate and inhibit YAP/TAZ activity.”.

Comments 3.      Line 60, YAP/TAZ and remains YAP/TAZ in the cytoplasm. I am not sure that “remain” is the correct work, please consider re-wording (e.g. retain).  

Response 3: Thanks a lot for pointing this out. We have revised this typo into ‘retain’.

Comments 4.      Line 78, Please consider adding a figure to show this signaling pathway as this will help the readers to understand the signaling pathway.

Response 4: Agree. We have, accordingly, added Figure 1 to emphasize this point. Please see Figure 1 in the revise manuscript.

Comments 5.      Line 119, the Authors mention that YAP is methylated, and this increases its oncogenicity by preventing it nuclear translocation. The Authors are asked to explain this further as they mentioned in lines 84 and 85 that YAP nuclear translocation activate target genes and target genes are mainly proliferative and survival genes (i.e. oncogenic). Then preventing YAP nuclear translocation should reduce oncogenicity not increasing. Please clarify.

Response 5: Thank you for your comment. Actually the oncogenic activity of YAP is dramatically inhibited after methylation, which is in accordance with lines 84 and 85 that YAP nuclear translocation activate target genes and target genes are mainly proliferative and survival genes. We have revised this mistake, please see Line 119-122.

Comments 6.      Lines 154 to 160, It is not clear for me as the Authors first indicate that impaired autophagy increases YAP but they conclude in the last non-referenced statement (lines 158 to 160), that YAP suppresses autophagy. This seems like vicious circle. Would the Authors be able to clarify this further? And add reference to the final statement (lines 158 to 160)?

Response 6: Thanks a lot for your comment. As you mentioned that, actually this is a vicious circle, impaired autophagy increases YAP and then, and as feedback, YAP is markedly upregulated can enhance drug resistance by suppressing autophagy-related cell death, which can cause further progression of liver cancer. We have modified this sentence to improve its readability, please see Line 160-162.

Comments 7.      Lines 164 to 167, The Authors describe the effects of TAZ knockdown in liver cancer cells, but these effects do not seem to align with the effects of TAZ knockout which causes YAP overexpression as described in lines 89 to 90. Would the Authors be able to clarify this point and whether knockdown and knockout have different effects?

Response 7: Thanks a lot for pointing this out. We admit that line 89-90 is an incorrect description and we have deleted it in the revised manuscript.

Comments 8.      Lines 190 to 192, the effect of hypoxia seems to be inducing YAP and its relocation to nucleus, so it is not clear for me how this would increase the sensitivity of HCC cells to irinotecan? Would the Authors be able to clarify?

Response 8: Thank you very much for your comment. Actually, hypoxia decreased the sensitivity of HCC cells to irinotecan, we have revised this typo. Please see Line 198-201 in the revised manuscript.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I would suggest the authors to change “pectrin beta” to “spectrin beta” in line 255 and correct the sentence in line 298 grammatically.  

 

Comments on the Quality of English Language

Dear Editor,

The manuscript “Targeting the Hippo-YAP/TAZ Signaling Pathway in Primary Liver Cancer Therapy” by Yina Wang and Liangyou Rui, has been revised thoroughly and can now be considered for publication. I would suggest the authors to change “pectrin beta” to “spectrin beta” in line 255 and correct the sentence in line 298 grammatically.  

Best regards,