Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The topic of the article, as well as the research itself and the results obtained, is relevant and important.
Fucosylation is a key driver and marker of colorectal cancer (CRC) progression at several levels: tumour growth, invasion–metastasis, stemness, immune escape, and therapy response.
Attachment of fucose to glycans changes during colorectal carcinogenesis and influences many steps of tumour development and progression. Multiple fucosyltransferases (FUT3, 4, 6, 7, 8, 9) are upregulated in CRC and modify receptors and adhesion molecules that control proliferation, migration, and survival.
However there are some essential comments.
1.     There is no description of the initial and final datasets, criteria for inclusion and exclusion, number of observations analyzed, methods for processing data gaps. Not specified in materials and methods, repository access path.
2.     Despite the considerable amount of information provided, there is no overall graphical representation of the generalized results of the study, the value of the fucosylation for colorectal cancer progression, tabular or graphical generalization of the dependencies of studied indicators on the degree of fucosylation.
3.    Need to check the literature list. Links 57, 58, 64 could not be found; such DOI does not exist. Link 58 may contain a misprint as a similar article with the same authors was found Okeley NM, Alley SC, Anderson ME, Boursalian TE, Burke PJ, Emmerton KM, Jeffrey SC, Klussman K, Law CL, Sussman D, Toki BE, Westendorf L, Zeng W, Zhang X, Benjamin DR, Senter PD. Development of orally active inhibitors of protein and cellular fucosylation. Proc Natl Acad Sci U S A. 2013 Apr 2;110(14):5404-9. doi: 10.1073/pnas.1222263110. Epub 2013 Mar 14. PMID: 23493549; PMCID: PMC3619284.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This article looks at the fucosylation dynamics as a critical determinant of cancer cell fate in colorectal carcinoma. Role of fucosylation as an integrative determinant of cancer cell fate in colorectal cancer (CRC) remains undefined. Here the authors investigated how sugar modifications on cancer cell surfaces, specifically fucosylation, influences tumor behavior and patient survival. The authors analysed transcriptomic and clinicopathological data from nearly 1,000 colorectal cancer patients and discovered that tumours fall into two distinct groups based on their fucosylation levels. The authors established tumor fucosylation as an integrative determinant of cancer cell fate in colorectal carcinoma, defining two fundamentally distinct tumour cell states with mutually exclusive engagement of invasion programmes, RTK signalling architectures, metabolic pathways, immune phenotypes, and drug resistance mechanisms. The findings from this article enable the independent prognostic value of fucosylation status, coupled with its integration of diverse biological programmes, and positions it as a powerful biomarker for risk stratification and personalised treatment decisions in colorectal cancer. Measuring fucosylation levels could help doctors predict which patients will have more aggressive disease and might guide treatment decisions, potentially matching patients with therapies most likely to work for their specific tumour type. The findings from this article lay a solid base to expand into more mechanistic and biochemical studies, to prove the links between fucosylation and the phenotypic features identified in this article and clinical studies to prove the utility of this biomarker. The authors have done a good job of highlighting the potential of fucosylation to be used as a biomarker in the field of cancer biology. However, I have some questions and concerns regarding some of the data.  I would recommend this article for publication after major revision.

 

One of the major issues with this article in general is the figure legends. In each figure I would recommend the author provide a panel number for each figure: For example: Figure 1A, 1B, 1C and so on! The authors refer to the specific figures as top left, top right etc. However, when there are multiple figure panels (example: figure 4), it becomes confusing. Also include these figure panel numbers in the figure legend.

For most part the figure and tables are referred at the very end of the paragraph, I would recommend the authors to refer to the specific figure at the end of the statement that talks about that data.

 

In the introduction section, Line 68-70, I believe that there is a reference missing for that statement. Include the right reference.

Results section: 3.1.1: Is the data in the first paragraph present in the form of table, if yes which table? There is no reference to any table or figure.

Line 276: Table1 should be Table 1.

In Figure 2: Mucinous tumors showed elevated fucosylation score, can this be also due to the # of samples. There are clearly less data points for the mucinous vs non-mucinous tumor.

In figure 4: In the figure legend the authors mention that “BRAF-mutant tumours also demonstrated reduced fucosylation relative to BRAF wild-type cases.” However, in the figure it seems it is the opposite and BRAF mutations demonstrate enrichment in high fucosylation tumors.

Figure 6: In figure 6 lowest right panel, some data points are not aligned and are confusing. For example, for metabolism of lipids, the high and low fucosylation datapoints are not next to each other. I would change it, so that it is consistent with all the other figures.

Did the authors look at the expression of various fucosylation genes in their dataset to see how these correlates with low and high fucosylation tumors and if they can identify any specific fucosylation genes that enriched in low vs high fucosylation?

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Thank you to the authors for making adjustments to the publication in line with my comments and adding informative illustrations.
However, there are still questions about the sample description. The authors only specified the total number of observations but did not specify their distribution among source databases. The initial number of observations is also not sufficiently described.

Author Response

Comment 1: Thank you to the authors for making adjustments to the publication in line with my comments and adding informative illustrations.
However, there are still questions about the sample description. The authors only specified the total number of observations but did not specify their distribution among source databases. The initial number of observations is also not sufficiently described.

Response 1: The statements in bold fonts have been added to sub-section 2.1 to clarify the distribution of cases across the datasets, including the initial numbers retrieved and those retained after filtering. These changes are highlighted in green in the revised manuscript.

2.1. Study Cohorts and Data Harmonisation‎

This study utilised transcriptomic and clinicopathological data from three independent colorectal ‎cancer cohorts. The Cancer Genome Atlas (TCGA-CRC) and Clinical Proteomic Tumor Analysis ‎Consortium (CPTAC2-CRC) datasets were accessed through publicly available repositories as ‎previously described [1,12]. An additional cohort, the Sidra–LUMC CRC cohort [13], was also included. ‎Only primary colorectal adenocarcinoma specimens with available transcriptomic and clinical ‎annotations were retained for analysis. A total of 995 primary tumour samples were initially retrieved across the three datasets, ‎comprising TCGA (n = 537), Sidra–LUMC (n = 348), and CPTAC2 (n = 110). Inclusion criteria comprised primary colorectal adenocarcinoma samples with available transcriptomic ‎profiles and corresponding clinicopathological annotations. Samples lacking key clinical variables or ‎with incomplete transcriptomic data were excluded from the analysis. Following quality control and application of inclusion and exclusion criteria, the final analytical ‎cohort consisted of 988 tumours, distributed as follows: TCGA (n = 534), Sidra–LUMC (n = ‎‎348), and CPTAC2 (n = 106).‎ Publicly available ‎datasets were accessed via the Genomic Data Commons (TCGA) and CPTAC data portals, while the ‎Sidra–LUMC dataset was obtained from previously published sources. Missing data were handled by ‎case-wise exclusion for analyses requiring complete variables.‎

Reviewer 2 Report

Comments and Suggestions for Authors

This review article looks at the fucosylation dynamics as a critical determinant of cancer cell fate in colorectal carcinoma. Role of fucosylation as an integrative determinant of cancer cell fate in colorectal cancer (CRC) remains undefined. Here the authors investigated how sugar modifications on cancer cell surfaces, specifically fucosylation, influences tumor behavior and patient survival. The authors analysed transcriptomic and clinicopathological data from nearly 1,000 colorectal cancer patients and discovered that tumours fall into two distinct groups based on their fucosylation levels. The authors established tumor fucosylation as an integrative determinant of cancer cell fate in colorectal carcinoma, defining two fundamentally distinct tumour cell states with mutually exclusive engagement of invasion programmes, RTK signalling architectures, metabolic pathways, immune phenotypes, and drug resistance mechanisms. The findings from this article enable the independent prognostic value of fucosylation status, coupled with its integration of diverse biological programmes, and positions it as a powerful biomarker for risk stratification and personalised treatment decisions in colorectal cancer. Measuring fucosylation levels could help doctors predict which patients will have more aggressive disease and might guide treatment decisions, potentially matching patients with therapies most likely to work for their specific tumour type. The findings from this article lay a solid base to expand into more mechanistic and biochemical studies, to prove the links between fucosylation and the phenotypic features identified in this article and clinical studies to prove the utility of this biomarker. The authors have done a good job of highlighting the potential of fucosylation to be used as a biomarker in the field of cancer biology. I thank the authors for making the recommended changes and answering the questions and hopefully these changes will strengthen the paper. I understand that not all of the analysis can be done for this paper and could be a future thing to do. I would recommend this article for publication in its current form.

Author Response

Final Comment: This review article looks at the fucosylation dynamics as a critical determinant of cancer cell fate in colorectal carcinoma. Role of fucosylation as an integrative determinant of cancer cell fate in colorectal cancer (CRC) remains undefined. Here the authors investigated how sugar modifications on cancer cell surfaces, specifically fucosylation, influences tumor behavior and patient survival. The authors analysed transcriptomic and clinicopathological data from nearly 1,000 colorectal cancer patients and discovered that tumours fall into two distinct groups based on their fucosylation levels. The authors established tumor fucosylation as an integrative determinant of cancer cell fate in colorectal carcinoma, defining two fundamentally distinct tumour cell states with mutually exclusive engagement of invasion programmes, RTK signalling architectures, metabolic pathways, immune phenotypes, and drug resistance mechanisms. The findings from this article enable the independent prognostic value of fucosylation status, coupled with its integration of diverse biological programmes, and positions it as a powerful biomarker for risk stratification and personalised treatment decisions in colorectal cancer. Measuring fucosylation levels could help doctors predict which patients will have more aggressive disease and might guide treatment decisions, potentially matching patients with therapies most likely to work for their specific tumour type. The findings from this article lay a solid base to expand into more mechanistic and biochemical studies, to prove the links between fucosylation and the phenotypic features identified in this article and clinical studies to prove the utility of this biomarker. The authors have done a good job of highlighting the potential of fucosylation to be used as a biomarker in the field of cancer biology. I thank the authors for making the recommended changes and answering the questions and hopefully these changes will strengthen the paper. I understand that not all of the analysis can be done for this paper and could be a future thing to do. I would recommend this article for publication in its current form.

 

Final Response: 

We sincerely thank the reviewer for the thorough evaluation of our manuscript and for the constructive feedback provided throughout the review process. We greatly appreciate the reviewer’s recognition of the novelty and significance of our work in establishing tumour fucosylation as an integrative determinant of cancer cell fate in colorectal cancer, as well as the acknowledgment of its potential clinical utility as a prognostic biomarker for risk stratification and therapeutic decision-making. We also appreciate the reviewer’s understanding regarding the scope of the current study and the need for future mechanistic and biochemical investigations. As highlighted, we agree that further studies will be essential to experimentally validate the links between fucosylation and the diverse phenotypic programmes identified, as well as to establish its clinical applicability. We are grateful for the reviewer’s recommendation for publication and for the insightful comments that have helped strengthen the manuscript.