Reviewer’s report
Title: Validity of biopsy-based drug effects in a diet-induced obese mouse model of biopsyconfirmed
NASH
Version: 0 Date: 13 Aug 2019
Reviewer: Nakade Yukiomi
Reviewer's report:
Please include all comments for the authors in this box rather than uploading your report as an
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, supporting
materials or other aspects of your report which cannot be included in a text format.
Please overwrite this text when adding your comments to the authors.
The authors showed that stereological validation of biopsy-based histopathology assessment of drug
treatment effects in a diet-induced obese mouse model of biopsy-confirmed NASH. The purpose of this
study is to examine how well biopsy-based quantitative image analyses correlate to stereology-based
whole-liver quantitative changes upon drug treatment. The authors concluded that an in-depth analyses
of  liver  homogeneity  demonstrated  that  drug  induced  improvement  in  liver  biopsy-assessed
histopathology is representative for overall liver effects assessed using stereology. Unfortunately, this
paper is very difficult to read. Therefore, it describes the evaluation of histological heterogeneity in the
liver in NASH and the comparison of the effects of the three drugs against it, but this is very difficult to
understand. In particular, the discussion section is very difficult to understand, so it is recommended to
rebuild it.
Major comments
GLP-1 agonist, INT-767, elafibronor are used in the pharmacotherapy intervention for NASH. The
reason why these drugs were used should be given. There is no explanation of these drugs in the
introduction or abstract, so it should be mentioned first. And please explain how the effectiveness of
these drugs against NASH is compared with past experiments.
Inflammation  is  assessed  by  Gal-3  and  fibrosis  assessment  by  col1A1,  but  why  did  other
immunohistological markers not be adopted? What was the actual inflammatory cell infiltration and
liver fibrosis? I think that it should be evaluated by SAF score (Beodssa et al, Hepatology, 2014) and
not NAS.。
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
Yes
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
Yes
Are you able to assess any statistics in the manuscript or would you recommend an additional
statistical review?
If  an  additional  statistical  review  is  recommended,  please  specify  what  aspects  require  further
assessment in your comments to the editors.
Not relevant to this manuscript
Quality of written English
Please indicate the quality of language in the manuscript:
Needs some language corrections before being published
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below. If
your reply is yes to any, please give details below.
I declare that I have no competing interest
I agree to the open peer review policy of the journal. I understand that my name will be included on my
report to the authors and, if the manuscript is accepted for publication, my named report including any 
attachments I upload will be posted on the website along with the authors' responses. I agree for my 
report to be made available under an Open Access Creative Commons CC-BY license 
(http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments which I do not wish to 
be included in my named report can be included as confidential comments to the editors, which will not
be published.
I agree to the open peer review policy of the journal
Reviewer’s report
Title: Validity of biopsy-based drug effects in a diet-induced obese mouse model of biopsyconfirmed
NASH
Version: 0 Date: 13 Aug 2019
Reviewer: Nakade Yukiomi
Reviewer's report:
Please include all comments for the authors in this box rather than uploading your report as an
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, supporting
materials or other aspects of your report which cannot be included in a text format.
Please overwrite this text when adding your comments to the authors.
The authors showed that stereological validation of biopsy-based histopathology assessment of drug
treatment effects in a diet-induced obese mouse model of biopsy-confirmed NASH. The purpose of this
study is to examine how well biopsy-based quantitative image analyses correlate to stereology-based
whole-liver quantitative changes upon drug treatment. The authors concluded that an in-depth analyses
of  liver  homogeneity  demonstrated  that  drug  induced  improvement  in  liver  biopsy-assessed
histopathology is representative for overall liver effects assessed using stereology. Unfortunately, this
paper is very difficult to read. Therefore, it describes the evaluation of histological heterogeneity in the
liver in NASH and the comparison of the effects of the three drugs against it, but this is very difficult to
understand. In particular, the discussion section is very difficult to understand, so it is recommended to
rebuild it.
Major comments
GLP-1 agonist, INT-767, elafibronor are used in the pharmacotherapy intervention for NASH. The
reason why these drugs were used should be given. There is no explanation of these drugs in the
introduction or abstract, so it should be mentioned first. And please explain how the effectiveness of
these drugs against NASH is compared with past experiments.
Inflammation  is  assessed  by  Gal-3  and  fibrosis  assessment  by  col1A1,  but  why  did  other
immunohistological markers not be adopted? What was the actual inflammatory cell infiltration and
liver fibrosis? I think that it should be evaluated by SAF score (Beodssa et al, Hepatology, 2014) and
not NAS.。
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
Yes
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
Yes
Are you able to assess any statistics in the manuscript or would you recommend an additional
statistical review?
If  an  additional  statistical  review  is  recommended,  please  specify  what  aspects  require  further
assessment in your comments to the editors.
Not relevant to this manuscript
Quality of written English
Please indicate the quality of language in the manuscript:
Needs some language corrections before being published
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below. If
your reply is yes to any, please give details below.
I declare that I have no competing interest
I agree to the open peer review policy of the journal. I understand that my name will be included on my
report to the authors and, if the manuscript is accepted for publication, my named report including any 
attachments I upload will be posted on the website along with the authors' responses. I agree for my 
report to be made available under an Open Access Creative Commons CC-BY license 
(http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments which I do not wish to 
be included in my named report can be included as confidential comments to the editors, which will not
be published.
I agree to the open peer review policy of the journal
Please review this paper.
user
Reviewer’s report
Title: Validity of biopsy-based drug effects in a diet-induced obese mouse model of biopsyconfirmed
NASH
Version: 0 Date: 13 Aug 2019
Reviewer: Nakade Yukiomi
Reviewer's report:
Please include all comments for the authors in this box rather than uploading your report as an
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, supporting
materials or other aspects of your report which cannot be included in a text format.
Please overwrite this text when adding your comments to the authors.
The authors showed that stereological validation of biopsy-based histopathology assessment of drug
treatment effects in a diet-induced obese mouse model of biopsy-confirmed NASH. The purpose of this
study is to examine how well biopsy-based quantitative image analyses correlate to stereology-based
whole-liver quantitative changes upon drug treatment. The authors concluded that an in-depth analyses
of  liver  homogeneity  demonstrated  that  drug  induced  improvement  in  liver  biopsy-assessed
histopathology is representative for overall liver effects assessed using stereology. Unfortunately, this
paper is very difficult to read. Therefore, it describes the evaluation of histological heterogeneity in the
liver in NASH and the comparison of the effects of the three drugs against it, but this is very difficult to
understand. In particular, the discussion section is very difficult to understand, so it is recommended to
rebuild it.
Major comments
GLP-1 agonist, INT-767, elafibronor are used in the pharmacotherapy intervention for NASH. The
reason why these drugs were used should be given. There is no explanation of these drugs in the
introduction or abstract, so it should be mentioned first. And please explain how the effectiveness of
these drugs against NASH is compared with past experiments.
Inflammation  is  assessed  by  Gal-3  and  fibrosis  assessment  by  col1A1,  but  why  did  other
immunohistological markers not be adopted? What was the actual inflammatory cell infiltration and
liver fibrosis? I think that it should be evaluated by SAF score (Beodssa et al, Hepatology, 2014) and
not NAS.。
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
Yes
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
Yes
Are you able to assess any statistics in the manuscript or would you recommend an additional
statistical review?
If  an  additional  statistical  review  is  recommended,  please  specify  what  aspects  require  further
assessment in your comments to the editors.
Not relevant to this manuscript
Quality of written English
Please indicate the quality of language in the manuscript:
Needs some language corrections before being published
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below. If
your reply is yes to any, please give details below.
I declare that I have no competing interest
I agree to the open peer review policy of the journal. I understand that my name will be included on my
report to the authors and, if the manuscript is accepted for publication, my named report including any 
attachments I upload will be posted on the website along with the authors' responses. I agree for my 
report to be made available under an Open Access Creative Commons CC-BY license 
(http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments which I do not wish to 
be included in my named report can be included as confidential comments to the editors, which will not
be published.
I agree to the open peer review policy of the journal
assistant