Title: SPOCK1 and POSTN are valuable prognostic biomarkers and correlate with tumor
immune infiltrates in colorectal cancer
Version: 1 Date: 03 Aug 2022
Reviewer's report:
This manuscript  discusses the possibility that SPOCK1 and POSTN expression may be
associated with tumor-infiltrating immune cells. However, I think that there are several problems
with the validation method and its interpretation.
Major points
1. in section Result 3.4, univariate Cox regression analysis for OS is performed; please also
show the results of multivariate COX regression analysis for OS.
2.In Fig. 9B, immunohistological staining is performed on cancerous and normal tissues with
target markers and blood cell markers. For cancer tissue, sections stained with target and
macrophage markers should show similar sections of the same specimen. The reader should
be interested in the location of POSTN and SPOCK1 expressing cells and tumor infiltrating
macrophages. This is also true for immunofluorescence assays.
Also, this figure alone does not address the question in section Result 3.7, "Above results, it is
not difficult to find that SPOCK1 and POSTN are co-expressed with CD68 and CD206 in colon
cancer, which premised speculated that SPOCK1 and POSTN may promote the poor prognosis
of colon cancer by promoting the polarization of M2-type macrophages”. It seems to be a slight
over-interpretation.
3. In Fig. 10, the correlation between SPOCK1 and POSTN expression and the expression of
immune checkpoint molecules is observed. Could you check the correlation between immune
checkpoint molecules such as PD-1 and TIM-3 (or microsatellite instability) and SPOCK1
(POSTN1) in immunohistological staining?
Also, the statement in Result 3.9, "These suggested that high expression of SPOCK1 and
POSTN might potentially regulate tumor immune evasion in CRC”, seems to be a slight over-
interpretation.
4. To support the claims made in this manuscript, it would be desirable to examine the prognosis
and response rates of patients treated with immune checkpoint inhibitors by SPOCK1 and
POSTN expression, if possible.
Title: SPOCK1 and POSTN are valuable prognostic biomarkers and correlate with tumor
immune infiltrates in colorectal cancer
Version: 1 Date: 03 Aug 2022
Reviewer's report:
This manuscript  discusses the possibility that SPOCK1 and POSTN expression may be
associated with tumor-infiltrating immune cells. However, I think that there are several problems
with the validation method and its interpretation.
Major points
1. in section Result 3.4, univariate Cox regression analysis for OS is performed; please also
show the results of multivariate COX regression analysis for OS.
2.In Fig. 9B, immunohistological staining is performed on cancerous and normal tissues with
target markers and blood cell markers. For cancer tissue, sections stained with target and
macrophage markers should show similar sections of the same specimen. The reader should
be interested in the location of POSTN and SPOCK1 expressing cells and tumor infiltrating
macrophages. This is also true for immunofluorescence assays.
Also, this figure alone does not address the question in section Result 3.7, "Above results, it is
not difficult to find that SPOCK1 and POSTN are co-expressed with CD68 and CD206 in colon
cancer, which premised speculated that SPOCK1 and POSTN may promote the poor prognosis
of colon cancer by promoting the polarization of M2-type macrophages”. It seems to be a slight
over-interpretation.
3. In Fig. 10, the correlation between SPOCK1 and POSTN expression and the expression of
immune checkpoint molecules is observed. Could you check the correlation between immune
checkpoint molecules such as PD-1 and TIM-3 (or microsatellite instability) and SPOCK1
(POSTN1) in immunohistological staining?
Also, the statement in Result 3.9, "These suggested that high expression of SPOCK1 and
POSTN might potentially regulate tumor immune evasion in CRC”, seems to be a slight over-
interpretation.
4. To support the claims made in this manuscript, it would be desirable to examine the prognosis
and response rates of patients treated with immune checkpoint inhibitors by SPOCK1 and
POSTN expression, if possible.
Please review this paper.
user
Title: SPOCK1 and POSTN are valuable prognostic biomarkers and correlate with tumor
immune infiltrates in colorectal cancer
Version: 1 Date: 03 Aug 2022
Reviewer's report:
This manuscript  discusses the possibility that SPOCK1 and POSTN expression may be
associated with tumor-infiltrating immune cells. However, I think that there are several problems
with the validation method and its interpretation.
Major points
1. in section Result 3.4, univariate Cox regression analysis for OS is performed; please also
show the results of multivariate COX regression analysis for OS.
2.In Fig. 9B, immunohistological staining is performed on cancerous and normal tissues with
target markers and blood cell markers. For cancer tissue, sections stained with target and
macrophage markers should show similar sections of the same specimen. The reader should
be interested in the location of POSTN and SPOCK1 expressing cells and tumor infiltrating
macrophages. This is also true for immunofluorescence assays.
Also, this figure alone does not address the question in section Result 3.7, "Above results, it is
not difficult to find that SPOCK1 and POSTN are co-expressed with CD68 and CD206 in colon
cancer, which premised speculated that SPOCK1 and POSTN may promote the poor prognosis
of colon cancer by promoting the polarization of M2-type macrophages”. It seems to be a slight
over-interpretation.
3. In Fig. 10, the correlation between SPOCK1 and POSTN expression and the expression of
immune checkpoint molecules is observed. Could you check the correlation between immune
checkpoint molecules such as PD-1 and TIM-3 (or microsatellite instability) and SPOCK1
(POSTN1) in immunohistological staining?
Also, the statement in Result 3.9, "These suggested that high expression of SPOCK1 and
POSTN might potentially regulate tumor immune evasion in CRC”, seems to be a slight over-
interpretation.
4. To support the claims made in this manuscript, it would be desirable to examine the prognosis
and response rates of patients treated with immune checkpoint inhibitors by SPOCK1 and
POSTN expression, if possible.
assistant