PONE-D-24-04415Activated Gab1 drives hepatocyte proliferation and anti-apoptosis in liver fibrosis via potential involvement of the HGF/c-Met signaling axisPLOS ONE

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PLOS ONE

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- National Institutes of Health

  

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: I Don't Know

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors have presented a well written and rigorously conducted experiment. Figures and tables have been provided to demonstrate the results and statistical analyses conducted. The conclusion obtained from the study provides important insight into the role of Gab-1 in the therapeutics of chronic liver disease.

Reviewer #2: PLoS ONE – PONE-D-24-04415 – Park et al.

The authors investigated the role of Gab1, an adaptor protein for various growth factor and cytokine receptors (including HGF, a hepatocyte growth factor), that is involved with cell differentiation and survival signaling pathways in the progression of liver fibrosis. Increased level of Gab1 is strongly correlated with poor prognosis in patients with hepatocellular carcinoma. Cellular and molecular biology methodology was used. Using hepatocytes isolated from humanized mice after acute viral infection, the authors studied signaling molecules related to the progression of liver diseases. A decreased level of pGab1 was detected simultaneously with an increased activation of TGF-β- related cell survival signaling pathways; TGF-β-mRNA expression was higher in infected cells compared to control. Because of the suggestion that apoptosis and proliferation signals were simultaneously activated in hepatocytes following viral infection, the authors hypothesized that Gab1 may participate in regulating the severity of liver disease progression by interacting with the TGF-β signaling pathway. Kinetic studies indicated that Gab1 expression was inversely related to the production of TGF-β during viral infection in hepatocytes. However, when using an established murine model of liver fibrosis induced by carbon tetrachloride treatment, Gab1 mRNA and relative protein levels increased in comparison to the control. Gab1 was thus suggested to play a role in progression of chronic liver disease by regulating cell proliferation. Using Gab1 knockdown cells, the results suggested that the expression of Gab1 is induced by HGF-c-Met signaling axis and that it is involved in increasing cell proliferation/survival and TGF-β expression. The authors concluded that there are differences between early and late host response during liver disease progression. In the early stages of liver disease, the TGF-β signaling pathway is enhanced while Gab1 activation is decreased; during chronic liver disease pGab1 and the expression of molecules involved in cell survival/proliferation pathways increase.

The subject of this manuscript is very interesting and the methodology was judiciously chosen to solve complementary aspects of the results. Certainly, further studies are required to fully understand the contradictory results on Gab1-mediated signaling pathway.

Materials and Methods – It was not mentioned how picrosirius-stained regions were measured. Equipment? Methodology?

Results – Lines 292-293 – Apparently, it cannot be assumed that mRNA expression of ERK has increased. No statistical significance was indicated in Figure 2. Similarly, increase in cleaved caspase-3 cannot be attributed because p = 0.31 is not accepted to mean statistical significance (Line 297).

Line 327 and Figure 3A – Apparently, a significant increase of Gab1 mRNA occurred until D1. Line 330 and Figure 3D – The same consideration applies to ERK.

Fig. 4 – E does not refer to mRNA expression. Figure elements should be described sequentially in the text. Description of D should precede E or restructure of the position of the figure elements should be undertaken.

Fig. 5 – There is something wrong with (C); the column named Merge shows only DAPI staining. Legend: There is no ***P< 0.001 in this figure

Fig. 6 – (E) Insert “Liver damage” over the line where “Early phase” and “Late phase” appear written. Legend: *P< 0.05 and ***P< 0.001 do not refer to this figure.

Minor:

. Standardize: min or minutes

. Lines 128, 130, 133, 134 … 3 mm, 10 min, 30 min, 100 µm, … respectively

. Line 137 – Insert “assay” after blue

. Lines 146, 147, 243 – The first time the commercial source is mentioned, please add city and country

. Line 154 – “mm2”

. Lines 160, 375,395,402 – typos

. Line 171 – “described [17]”

. Line 178 – a verb is missing

. Lines 246-247 – Here, (Sigma, Aldrich) is sufficient. City and country should be added to Sigma mention at line 147.

References – 1. Please abbreviate the name of the journal

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Reviewer #1: No

Reviewer #2: No

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Activated Gab1 drives hepatocyte proliferation and anti-apoptosis in liver fibrosis via potential involvement of the HGF/c-Met signaling axis

PONE-D-24-04415R1

Dear Dr. Park,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Mahmoud Kandeel

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: I Don't Know

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #2: With your revision, I have no pending questions. Please verify that the typos you did not identify moved to other lines in the revised text:

line 160 - "descried"

line 379 - "Hydorxylproline"

lines 399 and 406 - "hepa"

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If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

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