RHO Family GTPases in the Biology of Lymphoma
Round 1
Reviewer 1 Report
Abstract
The Small GTPases are divided into Ras Ran Rho Arf and Rab GTPases . Depending on some of the classification this could be 6 including Miro GTPases.
The first sentence of the abstracted in misleading and incorrect.
What included frequently mutated genes? This is usually from the Ras molecules of the Ras Superfamily
Introduction
Paragraph 1
The classification given here is unclear. Reference Number 1 actually gives the correct subfamilies of Rho with appropriate figures. This is not reproduced well in the manuscript.
The subfamilies of Rho include Rac, Cdc42, Rho, Rnd, RhoT and RhoBTB. RhoBTB can taken out as an atypical GTPase But the other families are clearly classified for Years.
Most part of paragraph 2 lacks appropriate references. Original references on Oncogenic processes including transformation and Cytoskeletal remodelling implicated in Cancer needs to be provided.
Line 50/51 : The specific gain and loss-of function mutations in specific Rho species needs to be added.
Again Line 53/54 : What are the tumour supressors?
Line 54/55: Its has been established how Rho GTPase pathway regulation through down stream kinases lead to changes in focal adhesion and oncogenic processes. Please clarify why you state this.
Regulation of RHO family GTPases and physiological functions
The first paragraph is quite general and does not add value to the review in terms of adding value to the topic of interest.
It would be helpful if the ‘Aberrant expression levels or mutations of GEFs, GAPs or GDIs lead to increased activation of RHO GTPase signalling cascades that in turn promote cancer initiation and progression’ could have been explained in terms of general loss of the GEF/GAP/GDI regulation that leads to Lymphoma with specific Rho EXAMPLES.
What are the specific atypical GTPases we are thinking about when describing them in paragraph 98-105? Its quite nonspecific and loses impact.
Some of this addressed in the final parts of the review with a new introduction to the subtopic but this loses coherence.
Functional and genetic alterations of RHO family GTPases in lymphoma
Rho GTPases are a group of molecules of about 6 different sub-families. The content in most of these section largely generalise them as a single group with common behaviour.
This is not the case and emerging literature shows this quite clearly for the last decade.
RHOA in the pathogenesis of lymphoma
Having given the mutation of RhoA in table 1 the focus here needs to be on what were the biological significance and which of those were driver mutations and overall the specific facts that points towards the compelling role of RhoA in Lymphoma Pathogenesis.
Adult T-cell leukemia/lymphoma
Most part of the content here completely lacks references.
Signalling networks or established pathways that lead to pathogenesis of Lymphoma through specific Rho GTPases such as RhoA, RhoH will be informative.
Author Response
Point-by-point response to Referees
Dear Editor,
Thank you for your interest and consideration of our manuscript Cells-508087 entitled “RHO Family GTPases in the Biology of Lymphoma” by Voena Claudia and Chiarle Roberto.
We appreciated the comments of the Referees that thoughtfully reviewed our manuscript and provided significant and helpful comments to further improve it in a revised form.
Based on this evaluation and on your editorial decision, we are now submitting a revised version of the manuscript in which we carefully evaluated and addressed all the issues raised by the Reviewers.
Reviewer #1
We thank the Reviewer for his/her suggestions. Here is a list of corrections and edits we made to address the Reviewer’s comments:
Abstract
The Small GTPases are divided into Ras Ran Rho Arf and Rab GTPases. Depending on some of the classification this could be 6 including Miro GTPases. The first sentence of the abstract is misleading and incorrect.
We agree that the first sentence was misleading and not properly stated. Thus, we corrected as follows: RHO GTPases are a class of small molecules involved in the regulation of several cellular processes and belong to the RAS GTPase superfamily.
What included frequently mutated genes? This is usually from the Ras molecules of the Ras Superfamily
We specified which genes belonging to the RAS family are frequently mutated in human cancer, i.e. K-RAS, N-RAS and H-RAS.
Introduction
Paragraph 1
- The classification given here is unclear. Reference Number 1 actually gives the correct subfamilies of Rho with appropriate figures. This is not reproduced well in the manuscript. The subfamilies of Rho include Rac, Cdc42, Rho, Rnd, RhoT and RhoBTB. RhoBTB can taken out as an atypical GTPase But the other families are clearly classified for Years.
The classification of Rho GTPases is still slightly variable depending on the different reviews published. Since this classification is not a topic of our review, we provided the classification accordingly to different recent reviews that were to the References.
- Most part of paragraph 2 lacks appropriate references. Original references on Oncogenic processes including transformation and Cytoskeletal remodelling implicated in Cancer needs to be provided.
We included original references to paragraph 2 as suggested.
- Line 50/51: The specific gain and loss-of function mutations in specific Rho species needs to be added.
We included more information regarding some examples of gain and loss of function mutations described in human cancers. See page 2 from line 51 to 70 of the revised manuscript.
- Again Line 53/54: What are the tumour suppressors?
We added which Rho GTPase is considered a tumor suppressor on page 2 lines 72-77 of the revised manuscript.
- Line 54/55: Its has been established how Rho GTPase pathway regulation through down stream kinases lead to changes in focal adhesion and oncogenic processes. Please clarify why you state this.
The statement on line 54/55 was referred to RHOA because most of the mutations identified in human lymphomas still lack a definitive mechanism of tumorigenesis. In fact, mouse models clearly show that the RHOA G17V mutation in T lymphocytes is not sufficient to induce lymphoma per se, in contrast to other known oncogenes. The RHOA G17V mutation induces T cell lymphoma only when combined with the loss of the epigenetic regulator TET2. This cooperation is still very poorly understood. Thus, alteration of RHO GTPase downstream kinases is not a sufficient mechanism of lymphomagenesis. We commented on this point in the revised manuscript.
Regulation of RHO family GTPases and physiological functions
- The first paragraph is quite general and does not add value to the review in terms of adding value to the topic of interest.
We included a general paragraph about RHO GTPases functions and regulation in order to help the readers not specialized in the field.
- It would be helpful if the ‘Aberrant expression levels or mutations of GEFs, GAPs or GDIs lead to increased activation of RHO GTPase signalling cascades that in turn promote cancer initiation and progression’ could have been explained in terms of general loss of the GEF/GAP/GDI regulation that leads to Lymphoma with specific Rho EXAMPLES.
We make detailed examples as requested in the revised manuscript.
- What are the specific atypical GTPases we are thinking about when describing them in paragraph 98-105? Its quite nonspecific and loses impact.
We specifically indicated which atypical GTPases we were referring to in paragraph corresponding to lines 122-127 in the revised manuscript.
Functional and genetic alterations of RHO family GTPases in lymphoma
- Rho GTPases are a group of molecules of about 6 different sub-families. The content in most of these section largely generalise them as a single group with common behaviour. This is not the case and emerging literature shows this quite clearly for the last decade.
This section was meant to provide a brief general introduction to set the tone for the following paragraphs in which the role of each RHO GTPase in human lymphomas has been described in details. We revised the paragraph to make clear that individual GTPases have unique properties in different cancers and we will describe in details only their role in lymphoma.
RHOA in the pathogenesis of lymphoma
- Having given the mutation of RhoA in table 1 the focus here needs to be on what were the biological significance and which of those were driver mutations and overall the specific facts that points towards the compelling role of RhoA in Lymphoma Pathogenesis.
We explained more in details the biology of RhoA alterations in T cell lymphoma.
Adult T-cell leukemia/lymphoma
- Most part of the content here completely lacks references.
To date only Nagata et al., Blood 2016 have reported and described mutations affecting RHOA in adult T-cell leukemia/lymphoma (ATL). We added another reference (Kataoka K et al., Nature Genetics 2015) to this section that reports the identifications of RHOA mutations in ATL in an integrated molecular study in total of 426 ATL cases. To our knowledge there are not other references relevant to this type of lymphoma.
- Signalling networks or established pathways that lead to pathogenesis of Lymphoma through specific Rho GTPases such as RhoA, RhoH will be informative.
We provided additional information about the regulation by RhoAof the mTOR signaling in T cell lymphoma and by RHOH of BCL6 expression in B cell lymphoma.
Reviewer 2 Report
The review, about the role of the small GTPases of the Rho family in the biology of lymphoma developed by Voena and Chairle, addresses a very important issue in the molecular biology of cancer. The review is well structured, well written and despite the amount of updated information presented, it is easy to read it, in fact, any reader not specialized in the field will be able to follow the review without problems
But before accepting it for publication and as minor points:
I would suggest that the authors add something about the type of post-translational modifications that these “typical” GTPases present under physiological conditions and what aberrant post-translational modifications can contribute to tumorigenesis.
And finally, on Page 5, lane 2, Rhoa G17V has to change to RhoA G17V.
Author Response
Reviewer #2
We deeply appreciate the comments and the overall evaluation of Reviewer #2.
We added a paragraph “on the type of post-translational modifications that “typical” RHO GTPases present under physiological conditions and what aberrant post-translational modifications can contribute to tumorigenesis” as suggested by the reviewer. A paragraph entitled “Post-transcriptional and post-translational regulation of RHO GTPases” has been added on page 3.
We corrected “Rhoa” into “RhoA” as indicated by the reviewer, on page 6 line 251 of the revised manuscript.
Round 2
Reviewer 1 Report
The current revision to the manuscript makes this more accurate and informative for the researchers in Rho GTPase and monomeric GTPase in general.
