The Clinicopathological Significance of BiP/GRP-78 in Breast Cancer: A Meta-Analysis of Public Datasets and Immunohistochemical Detection
, Isa Carneiro 2,3, João Lobo 2,3,4, Rui Henrique 2,3,4, Carmen Jerónimo 2,3,4 and Luisa Alejandra Helguero 1,*
Round 1
Reviewer 1 Report
1. The exclusion criteria in meta-analysis could be more rigorous. Papers with same cases, incomplete data structure, and insufficient data for the assessment of hazard ratios (HRs) and 95% confidence intervals (CIs) could be also list in the exclusion criteria.
2. Age, tumor treatment and patient's prognosis could also be included in the meta regression analysis of BIP.
3. Begg’s funnel plot and Egger’s test were performed to detect publication bias, but the results (figure of the funnel plot or P value of Egger’s test) were not present in the paper.
4. There are few samples for BIP immunohistochemistry test (only 26 cases) from IPO-P, leading to a low reliability of the results.
5. Some titles have a cross page problem (2. Materials and Methods, 3.2. Effect of that and metastasis on BiP expression in break cancer), some tables have a cross page problem (Table 5.), and some pictures have a layout problem (Figure 4.)
Author Response
"Please see the attachment."
Author Response File: 
Author Response.docx
Reviewer 2 Report
Interesting findings and overall a reasonable manuscript. An article whose findings are important to those with closely related research interests. The paper is well written and correct from a methodological point of view. Many abbreviations make this manuscript difficult to read; please avoid nonessential abbreviations
Author Response
Reviewer 2
Interesting findings and overall a reasonable manuscript. An article whose findings are important to those with closely related research interests. The paper is well written and correct from a methodological point of view. Many abbreviations make this manuscript difficult to read; please avoid nonessential abbreviations.
Author response: We appreciate the positive comments. The Reviewer´s suggestion was taken into consideration and the nonessential abbreviations were excluded in this version of the manuscript.
Reviewer 3 Report
In this manuscript by Inês Direito et al, the authors investigated The clinicopathological significance of BiP/GRP-78 in breast cancer.
Comments and suggestions:
1. Abstract and simple summary overlap as ideas. I suggest synthesizing them only in an abstract
2. All abbreviations must be explained the first time they appear in the text, including the abstract. Revise it.
3. Introduction add more data about the clinical signs of breast cancer.
4. Figures 6,7 add more elements of histopathological description, what tissues are there and what changes there are.
What is the novelty of this paper?
What perspectives for human health does this MS have?
Consider revision accordingly.
Author Response
Reviewer 3
Author response: We appreciate the time taken to review our manuscript and the constructive critics, which have helped us improve this work. Please find our responses highlighted in blue in the manuscript text.
In this manuscript by Inês Direito et al, the authors investigated The clinicopathological significance of BiP/GRP-78 in breast cancer.
- Abstract and simple summary overlap as ideas. I suggest synthesizing them only in an abstract
Author response: We agree with the Reviewer and had initially included the simple summary as requested by the journal. However, since the simple summary is optional, we have deleted it in this version of the manuscript.
- All abbreviations must be explained the first time they appear in the text, including the abstract. Revise it.
Author response: All the nonessential abbreviations were excluded in this version of the manuscript and all the abbreviations included were explained the first time they appeared.
- Introduction add more data about the clinical signs of breast cancer.
Author response: We are not sure our interpretation of this comment is correct, please let us know if we misinterpreted the Reviewer´s suggestion. Regarding clinical signs of breast cancer (node, breast enlargement, etc..), we think this is too general and not related with our aim. Since it is out of the scope we have not included these ideas in the introduction. An alternative reading to this comment is that there needs to be more information regarding the association of UPR and clinical factors. In this case, we agree, and have now updated the following sentence (pg. 2): “Independent studies have also shown an association of UPR proteins with breast cancer clinical factors, such as lymph node metastasis, estrogen receptor positivity or decreased overall survival”.
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- Figures 6,7 add more elements of histopathological description, what tissues are there and what changes there are.
Author response: the following information was added in the legends of the figures: “Arrows indicate cytosolic BiP expression while arrowheads show BiP nuclear localization in neoplastic cells. All pictures were from the same invasive ductal carcinoma case as seen in the corresponding H&E slide showing irregular and angulated tubules infiltrating the breast stroma.”
5.What is the novelty of this paper?
Author response: The manuscript presented three different approaches and, in each one of them, there were new results presented. To clearly state the novelty of this work, the following sentence was added in the discussion: “To the best of our knowledge, this is the first study analysing the effect of hormone therapy in BiP positivity in human breast cancer tissue” (pg. 18). In addition, the most significant new findings are now summarized in the conclusions section: “Meta-analysis of publicly available RNA-seq and proteomics datasets of human breast cancers disclosed, for the first time, an association of BiP differential expression with immune signatures and a differential expression of BiP mRNA according to breast cancer molecular subtypes. The systematic review and meta-analysis of immunohistochemical detection of BIP identified for the first time that ER and HER2 are factors that contribute to BiP positivity. This metanalysis also confirmed that BiP positivity is significantly associated with recurrence free survival. The immunohistochemical detection of BiP in two small cohorts of luminal breast cancers disclosed a novel finding, the effect of endocrine treatment on BiP subcellular localization. and confirmed that increased BiP expression is associated with metastasis and HER2 expression.”
6.What perspectives for human health does this MS have?
Author response: In orther to clarify the perspectives, the following sentence was added to the conclusion: “In summary, BiP/GPR-78 has been implicated in response to stress caused by nutrient deprivation, hypoxia or resistance to chemotherapy in breast cancer. An active UPR thus provides a survival advantage to cancer cells over normal cells and the analysis presented in this manuscript showed the association between BiP expression and HER2 and basal molecular subtypes (usually more aggressive than luminal-like tumors), also its associa-tion with metastasis and short relapse free survival highlights the prognostic potential of BiP for breast cancer as an indicator of poorer outcome”
Round 2
Reviewer 3 Report
Accept
