Review for 10.1186/s12876-021-01713-9 (2021)

Reviewer’s report
Title: 
Expression of Cyr61 is associated with Clinical Course in Patients with
Crohn’s Disease
Version: 
1 
Date: 
25 Oct 2020
Reviewer's report:
The study entitled "Expression of Cyr61 is associated with Clinical Course in
Patients with Crohn's Disease" aimed to quantify protein and gene expression of
Cyr61 in CD biopsies compared to normal mucosa from controls. Age- and sex-
matched groups (each n=43) were selected for comparison using a propensity
score. Samples were extracted from FFPE blocks stored in the pathological
archive and collected in 2018-2019. Immunohistochemistry (IHC) was used to
evaluate the protein expression and a semi-quantitative score was calculated
based on staining intensity (low or high) and percentage of stained area.
Depending on the obtained score, patients were distributed in 3 groups (tertiles)
and their clinical outcome (recurrence) was evaluated. 
Results showed higher expression of Cyr61 in CD patients compared to controls.
However, worse clinical outcome was associated with lower Cyr61 expression. 
Several important issues and limitations should be addressed before publication. 
METHODS:
1. Methods should be better described. First, site (ileal or colonic) and
inflammatory activity of CD biopsies included in the analysis should be reported.
Site of control biopsies should be specified too. 
2. The evaluation of the staining intensity is subjective, even when done by an
experienced pathologist. It could be better to have the opinion of at least 2
different readers. 
3. Did you use an automatic calculator of the percentage of the stained area? Did
you use a specific program or application, such as ImageJ? Please, specify. 
4. RNA used for gene expression was obtained from 11 active and inactive CD
biopsies already used in a previously published paper: were paired samples
(active and inactive in the same patients?) Why only 11 among 32 patients
described in the previous study were selected? And how did you select the
samples to be included in this new study? Methods and results on gene
expression should be mentioned in the abstract too. 
5. Clinical and endoscopic disease activity scores (CDAI, SES-CD and CDEIS)
were measured at the time of colonoscopy. Did you retrieve these scores from
medical records? Or did you calculate them retrospectively? In the latter case, it
could be very imprecise and not methodologically correct (CDAI can be
calculated only prospectively, because it requires the collection of daily status of
the patient for 7 days). Also, retrospective calculation of the endoscopic scores
(SES-CD and CDEIS) may be very approximate. Such important limitations
should be mentioned in the discussion. 
RESULTS
1. second page, line 14: please, change "higher" with "lower".
2. Figure 1 should be mentioned in the results and not in the methods. Moreover,
it should be more appropriate to show representative images of control and CD,
and not 2 such different pictures both from CD patients. It is confusing. 
DISCUSSION
1. please, add a section on study limitations
2. possible explanation of study finding is not mentioned. Which should be the
role of Cyr61 in CD pathogenesis? Your results suggest that Cyr61 increases in
CD, but it is reduced in patients with future relapse. How do you explain such
conflicting findings? 
CONCLUSION
1. Based on the methods and the results of this study, the sentence "in the future,
the possible therapeutic role of Cyr61 should be considered" is completely
inappropriate. More detailed or functional experiments were not included in your
study and increased Cyr61 expression (as presented in this paper) may be just a
consequence of inflammation and may not have any active role in pro-
inflammatory pathways and pathogenesis. 
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Declaration of competing interests
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Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
No
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
Yes
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
Yes