Modelling 3D Tumour Microenvironment In Vivo: A Tool to Predict Cancer Fate

Round 1

Reviewer 1 Report

In this brief report by Marines et al., the Authors present a methodology to visualize in 3D live imaging the interaction between tumor microenvironment and cancer cells in zebrafish model.

The report is well written and interesting, however the used tools have already been used to examine in 3D the behavior of cancer cells in zebrafish. Due to that, a major concern should be addressed by the Authors, before a publication on Current Issues in Molecular Biology can be granted:

MAJOR CONCERN:

1.    The methodology described in the report can not be proposed as a novelty because the are works in literature in which the same softwares have already been used to visualize zebrafish in 3D. Moreover, that workflow used to generate fluorescent lines and to prepared larvae has already been detailed. Thus, the authors must stress this limit of the study and not report it as a new methodology.

Overall, major revision is required.

Author Response

We thank our referee for these positive comments and for appreciating our experimental approaches.

Regarding the comments of our article :

We do not minimize the work of our colleagues; on the contrary, we build on their approaches, as cited in our manuscript:

Gajewski, T. F., Schreiber, H. & Fu, Y. X. Innate and adaptive immune cells in the tumor microenvironment. Nature Immunology 14, 1014–1022 (2013).

Huang, S. et al. Identification of Immune Cell Infiltration and Immune-Related Genes in the Tumor Microenvironment of Glioblastomas. Front. Immunol. 11, 2708 (2020).

Wang, J. et al. Novel mechanism of macrophage-mediated metastasis revealed in a zebrafish model of tumor development. Cancer Res. 75, 306–315 (2015).

Hong, S. M. et al. Three-dimensional visualization of cleared human pancreas cancer reveals that sustained epithelial-to-mesenchymal transition is not required for venous invasion. Mod. Pathol. 33, 639–647 (2020).

Hamilton, L., Astell, K. R., Velikova, G. & Sieger, D. A Zebrafish Live Imaging Model Reveals Differential Responses of Microglia Toward Glioblastoma Cells In Vivo. doi:10.1089/zeb.2016.1339.

We agree that Imaris has already been used to image the zebrafish embryo in 3D. Protocols for staining and cell transplantation in zebrafish and their imaging have also been described in the literature by our colleagues.

However, for the first time, we described in detail a protocol for simultaneous visualization and monitoring of different cell populations that make up the cancer tumor microenvironment. We illustrated for the first time the collaboration of macrophages and vessels in the progression of cancer cells, in vivo, in 3D and 4D.

Since our publication in BioRxiv, our article has been downloaded several hundred times, demonstrating the interest of our protocol and approach for the scientific community: https://www.biorxiv.org/content/10.1101/2021.09.23.461461v1

We have taken your comments into account by deleting the words "For the first time", and "state of the art", as you will read in our revised version.

We would like to thank you once again for the time you have devoted to reading and reviewing our article, and we hope that this moderation will enable our article to be accepted.

Yours sincerely,

Karima Kissa.

Reviewer 2 Report

The manuscript entitled "Modelling 3D tumour microenvironment in vivo: a tool to predict cancer fate," authored by Marines et al. showcases a state-of-the-art approach to visualizing the interactions between tumour microenvironment and cancer cells using 3D live imaging. The 3D model presented in the study, which was conducted in zebrafish, can potentially be a valuable tool for not just studying patient-derived xenografts, as the authors have mentioned, but also for assessing the efficacy of novel cancer drugs. I highly recommend that this manuscript be published in CIMB, given its innovative methodology and potential for advancing cancer research.

Author Response

We thank our referee for these positive comments and for appreciating our experimental approaches presented in this manuscript.

Round 2

Reviewer 1 Report

In my opinion, after the changes applied, the manuscript is now ready to be published.