Title: CD63-positive extracellular vesicles are potential diagnostic biomarkers of pancreatic
ductal adenocarcinoma
Version: 2 Date: 02 Dec 2021
Reviewer's report:
In this Manuscript, Odaka et.al. reported the quantitative measurement of serum level CD63+
EVs and platelet derived EVs (CD41+and CD61+) in PDAC patients and healthy controls and
evaluated their potential use as biomarkers for PDAC.
This is an interesting study however the analyses have been done using small cohorts.
Specifically, the small number of PDAC patients in the cohort 2 and cohort 3 are likely to limit
analysis necessary to reach conclusions from the observed changes. Overall, to further strengthen
the manuscript, authors need to address some comments herein.
Comments:
1. It is important to determine the purities of the EVs in general. In the manuscript, patient serum-
and platelet-derived EV characterizations have not been done to detect their morphology and
sizes by transmission electron microscopy and by nano sight analysis respectively.
2. Authors described in the text as ‘universal CD63+ EVs’. EVs carrying tetraspanin CD9 or
CD81 form in the endosome but not CD63. Therefore, ‘universal EV’s are those which carries
CD63 together with at least one other tetraspanin (CD9 or CD81) that may represent to endosome
derived EVs (PMID: 26858453). Did authors detect other EV-associated tetraspanins e.g, CD9
or CD81 in the patient serum? It is suggested that either remove the ‘universal’ word from the
text or screen for another tetraspanin in addition to CD63.
3. In the result section page 13, line 154, authors described the combinational use of CD63+EVs
and CA19-9 as there were low correlation coefficient between the two. According to the
description in the method section, the detection assays of CD63+EVs and CA19-9 were
different. Therefore, it is unclear that how the values were combined and what was the rationale
behind it. Authors should explain that in details.
4. Authors examined the tumor burden on the serum levels EVs in two cohorts after surgery.
The number of the patients were too few to draw a strong conclusion that universal and platelet
derived EVs ‘reflect the tumor burden before and after surgery’.
Additional examples are necessary.
5. Page 13, line 151: Please correct the value from 0.853 to 0.858
6. Fig. 1: Y-axes of a, b and c should be in the same scale.
7. Fig. 4: Y-axes of b, c and d should be in the same scale.
Title: CD63-positive extracellular vesicles are potential diagnostic biomarkers of pancreatic
ductal adenocarcinoma
Version: 2 Date: 02 Dec 2021
Reviewer's report:
In this Manuscript, Odaka et.al. reported the quantitative measurement of serum level CD63+
EVs and platelet derived EVs (CD41+and CD61+) in PDAC patients and healthy controls and
evaluated their potential use as biomarkers for PDAC.
This is an interesting study however the analyses have been done using small cohorts.
Specifically, the small number of PDAC patients in the cohort 2 and cohort 3 are likely to limit
analysis necessary to reach conclusions from the observed changes. Overall, to further strengthen
the manuscript, authors need to address some comments herein.
Comments:
1. It is important to determine the purities of the EVs in general. In the manuscript, patient serum-
and platelet-derived EV characterizations have not been done to detect their morphology and
sizes by transmission electron microscopy and by nano sight analysis respectively.
2. Authors described in the text as ‘universal CD63+ EVs’. EVs carrying tetraspanin CD9 or
CD81 form in the endosome but not CD63. Therefore, ‘universal EV’s are those which carries
CD63 together with at least one other tetraspanin (CD9 or CD81) that may represent to endosome
derived EVs (PMID: 26858453). Did authors detect other EV-associated tetraspanins e.g, CD9
or CD81 in the patient serum? It is suggested that either remove the ‘universal’ word from the
text or screen for another tetraspanin in addition to CD63.
3. In the result section page 13, line 154, authors described the combinational use of CD63+EVs
and CA19-9 as there were low correlation coefficient between the two. According to the
description in the method section, the detection assays of CD63+EVs and CA19-9 were
different. Therefore, it is unclear that how the values were combined and what was the rationale
behind it. Authors should explain that in details.
4. Authors examined the tumor burden on the serum levels EVs in two cohorts after surgery.
The number of the patients were too few to draw a strong conclusion that universal and platelet
derived EVs ‘reflect the tumor burden before and after surgery’.
Additional examples are necessary.
5. Page 13, line 151: Please correct the value from 0.853 to 0.858
6. Fig. 1: Y-axes of a, b and c should be in the same scale.
7. Fig. 4: Y-axes of b, c and d should be in the same scale.
Please review this paper.
user
Title: CD63-positive extracellular vesicles are potential diagnostic biomarkers of pancreatic
ductal adenocarcinoma
Version: 2 Date: 02 Dec 2021
Reviewer's report:
In this Manuscript, Odaka et.al. reported the quantitative measurement of serum level CD63+
EVs and platelet derived EVs (CD41+and CD61+) in PDAC patients and healthy controls and
evaluated their potential use as biomarkers for PDAC.
This is an interesting study however the analyses have been done using small cohorts.
Specifically, the small number of PDAC patients in the cohort 2 and cohort 3 are likely to limit
analysis necessary to reach conclusions from the observed changes. Overall, to further strengthen
the manuscript, authors need to address some comments herein.
Comments:
1. It is important to determine the purities of the EVs in general. In the manuscript, patient serum-
and platelet-derived EV characterizations have not been done to detect their morphology and
sizes by transmission electron microscopy and by nano sight analysis respectively.
2. Authors described in the text as ‘universal CD63+ EVs’. EVs carrying tetraspanin CD9 or
CD81 form in the endosome but not CD63. Therefore, ‘universal EV’s are those which carries
CD63 together with at least one other tetraspanin (CD9 or CD81) that may represent to endosome
derived EVs (PMID: 26858453). Did authors detect other EV-associated tetraspanins e.g, CD9
or CD81 in the patient serum? It is suggested that either remove the ‘universal’ word from the
text or screen for another tetraspanin in addition to CD63.
3. In the result section page 13, line 154, authors described the combinational use of CD63+EVs
and CA19-9 as there were low correlation coefficient between the two. According to the
description in the method section, the detection assays of CD63+EVs and CA19-9 were
different. Therefore, it is unclear that how the values were combined and what was the rationale
behind it. Authors should explain that in details.
4. Authors examined the tumor burden on the serum levels EVs in two cohorts after surgery.
The number of the patients were too few to draw a strong conclusion that universal and platelet
derived EVs ‘reflect the tumor burden before and after surgery’.
Additional examples are necessary.
5. Page 13, line 151: Please correct the value from 0.853 to 0.858
6. Fig. 1: Y-axes of a, b and c should be in the same scale.
7. Fig. 4: Y-axes of b, c and d should be in the same scale.
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