Antagonism of Estrogen Receptor α-Driven Transcription Mediated by AP-1 in Breast Cancer Therapy
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
In this manuscript the author revealed that ERα conformal changes reflected by a temperature dependence of ligand binding affinity is an index of the ability of a ligand (estrogen, SERM or SERD) to modulate ERE and AP-1 transcriptions in MCF7 cell line. The manuscript is well written, clear organized. The layout content is short and concise. The conclusion is well supported by the results. Also the author had a comprehensive discussion about this manuscript and the future prospective directions. I would recommend publish this manuscript.
Minor suggestions:
(1) Line 34-35, check the sentence structure.
(2) Line 98, missing period.
(3) Line 231-232, check the sentence structure.
(4) Line 331, there were double periods.
Author Response
Thank you for these " supporting "comments.
Minor suggestions were taken ito account
L34-35 ... transciption procedures induced by its ligands. / L98 ERE transcriptions / L231-232.. was observed in parallel / L331 ...proliferation of MCF7 cells ( ) indiceated their perfect relevance
Reviewer 2 Report
Comments and Suggestions for Authors
Guy Leclercq in ” Antagonism of AP-1-Assisted on ERE-directed Transcriptions of Estrogen Receptor α, a
Factor to be taken Into Account in the Therapy Against Breast Cancer Endocrine Resistance”, summarized
recent findings about a few estradiol derivates bearing substituents in its position 11β that might contribute
not solely the development of drugs to abrogate this unfortunate issue but that may be also helpful to identify
molecular aspects of this receptor resistance in order to elaborate other therapeutic approaches.
The article paper is well written and very original. It is an in-depth and very articulated study. Only minor
revision is suggested as follows:
in each graph the standard deviation bar must be indicated.
Comments on the Quality of English Language
Minor editing of English language required
Author Response
Thank you for your report stressing The interest of our investigation. Text has been reviewed and therafter submited to artificial intelligence .
Computer Drawing system as inoperative : SD were globary recorded in the legends of the figure to state their reprouctibility
Reviewer 3 Report
Comments and Suggestions for Authors
Dear Author,
The review article titled "Antagonism of AP-1-Assisted on ERE-directed Transcriptions of Estrogen Receptor α, a Factor to be taken Into Account in the Therapy Against Breast Cancer Endocrine Resistance" by Guy Leclercq was described well.
One crucial aspect of breast cancer development involves the intricate interplay between estrogen signaling and gene transcription, particularly mediated by the estrogen receptor alpha (ERα). Among the regulatory factors in this process, two significant players emerge: AP-1 (Activator Protein 1) and ERE (Estrogen Response Element). In this exploration, the author, aim to unravel the critical roles of AP-1-assisted and ERE-directed ERα transcription in breast cancer. These mechanisms hold the key to understanding how estrogen signaling, orchestrated by ERα, impacts the behavior and proliferation of breast cancer cells.
Minor corrections:
1. Figure 1 is not good color contrast.
2. Figure 2 (A, B, C), bar graph is too small, and whether it is significant or not significant should be mentioned in the figure itself.
3. Figure 4, some of the texts are in bold, some of them are not in bold and turnover spelling mistake ( r is missing).
Author Response
Thanks for your comments revealing your interest.
Fig 1 : colors were modified to improve contrast.
Unfortunately our Drawing systemfailed to include SD ( impossilility of visualistion especially in FIf . Hence, reproducibbility was recorded in the legends of all figures (SD <x).
fig 4: Remark will be notified to Editor / Request for a change ( as for the table)
Reviewer 4 Report
Comments and Suggestions for Authors
The main outstanding first impression is that the article requires extensive quality controls, throughout the entire manuscript. It appears like having been produced in a hurry, with completely unnecessary and avoidable typos that could be spotted upon a single round of proofreading.
There are also numerous simple formatting errors, like missing or too many spaces between words and marks, at the end of sentences, everywhere. It is also striking that commas are often completely missing in entire paragraphs, replaced by extra empty spaces. So one has a hunch that there should be commas there, maybe, just that we dont see them.
Next, there are terms that are - to say the last - unusual. Like what are " ERα-mediated transcriptions procedures" (line 34 and elsewhere). Everyone knows what is mant, but still, it somehow appears odd. There are many such expressions that sometimes appear unnecessarily convoluted and "posh", complicated wording that's not even strictly scientific but borders on poetry.
This even applies to the title: "Antagonism of AP-1-Assisted on ERE-directed Transcriptions 1 of Estrogen Receptor α, a Factor to be taken Into Account in the 2 Therapy Against Breast Cancer Endocrine Resistance". This is, to say the least, the most unusual title I have encountered in a while because it takes a philosophy masters to decipher what it really means. It can be possibly said much more directly and in an unconvoluted fashion: "Antagonism of Estrogen-receptor driven transcription mediated by AP-1 in the therapy of breast cancers" or something like that? The title should be REALLY a clear statement, followed by the abstract; but both are not simple to decipher.
Then, many of the things pointed out in the first 2 pages are almost ancient textbook knowledge, for example, everything outlined in lines 43 - 55; that can be found in every textbook on genetics in the section "transcriptional regulation", usually with a nice illustration (which is missing here); and I am simply not sure if its needed here? In addition, there is essentially no clearly defined hypothesis and no future direction outlines in large parts of the manuscript. The reader gets the feeling this might be a review article; but checking again, no it seems to be original work; but where is it? These details outlined in the intro, especially those that are required to understand the following sections, definitely would require a schematic drawing or any illustration that brings things into context. There are plenty of bar graphs in this article but nothing that holds it together.
Then, suddenly, the manuscript gets very chemical. Here, the reader realizes that this is not a review but original work.
There are some graphical illustrations in the supplemental data, which are, however, of extremely poor quality and appear to be taken from some really olf pdf files or textbooks; definitely NOT modern day illustration quality levels; and they cannot be accepted in this form.
The figure legends of the bar graphs shown, for example, in Fig. 1 lack necessary information, as illustrated below:
Figure 1. ERb conformational change : No direct influence on down-regulation. Estrogen - induced ERE Transcription ( ligands : E2 / 11β-Ε2CCH at 0.1nM ).
It remains completely unclear what are we looking at? What is down-regulated; is it transcription? What are the ligands? Are the graphs now showing conformational changes, or down-regulations of something? Many questions, few answers! This is not acceptable and not professional writing.
By now, the authors gets the impression that maybe we are lookinginto original work, not a review article. But the typical elements of original articles are missing: there is no materials & methods section, there is no results section.
There are also no clear leading hypotheses what is the author trying to show, which problem is he trying to solve, with this work?
There is also no section in "discussion" that wold take up these hypotheses again and discuss if and how they may have been validated or rejected; no classic scientific writing here in place.
Meanwhile, the illuastrations and especially, the figure legends get even more cryptic. An example here from Figure 3:
"Figure 3. 11β - fulvestrant : AP-1 Τranscriptional Activities (MTLN cells) Mean of very reproductible data ( SD < 10%)"
Again, an endless list of questions, and uncertainties: What are MTLN cells, why have they not been described earlier? What are "very reproductable" data? And since when is 10% standard deviation (SD?) considered very reproducible? If anything, there is a 1% or 5% cutoff usually in place, in most analyses and statistics. What exactly is done with fulvestrant? How long was it used, which concentration(s) ? (okay, log molarity may be accepted but still, HOW LONG?)
It may be possible to understand what the author wants to tell us; but then;I dont really have the time to decipher largely cryptic texts! I have better things to do...
Comments on the Quality of English Language
quite difficult to understand, needs a lot of work
Author Response
First at all, I wish to stress that I strongly regret the impression of "a hurry" submission that might be ascribed to a precceding trensient but strong cardiac arithmy related to a ge-related cordiac disease (soon 80 yo).
Concerning the title of the manuscript, I totally agree that the one already proposed failed to cover its multi facets. Morever, it is not a "review " ( mention selected by MDPI) but an original investigation . Hence , I sbstituted this title by the one proposed in your comments since it fits largely better.
Now, with regard to the presentation of manuscript and related data in the context of breast cancer biology and treatment it should be kwown that it resultsfrom a request fom Kyra Yan for a submission devoted the present Special Issue. She accepted the present topic of which a large part of informations / data had not been pubhished ; for me, in view of my age, it was a chance to deliver the underlying message of this study, largelly commened in the concluding remarks. The vairous chemical, endocrine, molecular , thermodynamical aspects of the proposed work fited for her, giving rise to the present" bi-facets" manuscrpt which contains a review as first chapter to inform investigors unfamiliar with all these discipines . A presentention appreciated by the 3 other reviewers.
As mentioned, title of the manuscript has been changed, it refers to "driven" transcription. This terminology was used in the text ( L34) instead of mediated transciption sbjected to criticism.
Systematic responses to all comments of the text, legends of ligures , qualtiy of drawings .. would be quite fastidious . I did my best ! I wish nevertheless notify that a Material an Method section including the origin and properties of cells ( especallly MTLN ) was already present in the original submission ( 4.1 Experimenta Procedures ) localized in a postion preceeding the description of the exerimental data. Drawings for the Suppl Section were obviousy of sufficient quality ; text provides the references of publications fom which they are taken .
Comparison of previous and present manuscipts would facilitate the undersaning of my objective, taking into account of some styillistic changes not requested . Text was overviewed by an artificial inteligence . Request for a revion of table was adressed to the managing Editor
Concering reproducibility of all experiments ,SD were in recorded in the legends . For he one specifically adressed : only 2 valeus<10, all others<5.
Round 2
Reviewer 4 Report
Comments and Suggestions for Authors
The manuscript has undergone some improvements, but the main concern is still the lack of recognizable structure. I assume the author aimed for a "hybrid manuscript" but then it should really be obvious that this is what it is. It may be considered a heroic deed by the author to provide a manuscript thats "something different" and "stands out from the crowd", but then it should also contain some outstanding content.
So maybe there is a solution to this, but I think the author has to come half way and show some willingness to standardize this manuscript a bit more so it gets a recognizable format, and as such, is then truly informative.
I also still find a large amount of grammar issues, especially now with the construction of sentences, which are not acceptable, and should be fixed. I assume an AI program could help; the author claims this has been done but I can clearly see that it wasnt (manuscripts modified by AI-language models like ChatGPT do not contain any typos or grammatical issues; and they are still there). But that can still be done.
Then, it should be clarified that it is (as the authorr states) a manuscript of hybrid nature - partly review, partly data. Its not mentioned anywhere.
Next, there should still be a recognizable structure; every document needs this, and there is no "classic" structure of any kind in any of the 2 parts of the manuscript. If there are results, they should be clearly in a "results" section - but there is no results section.
If experiments are done, we do need a "materials & methods" section. Give it a different name, if you must - but it has to be explained somehow which cells were used, how they were treated, which chemical compounds and modifications thereof were used, who synthesized them, what is their structure - all of this is still missing. Then, the figure legends would also be much less cryptic: if the experiments were described somewhere, it should be easy to understand whats actually shown in the figures. But it isnt.
My concerns about interpunctation, lack of commas, missing or too many spaces between words and sentences are still all valid, they have not been removed. These are basic quality issues I would assume are a basic standard for manuscript submission. I do miss them here.
I am probably repeating myself here, but in the 1st part, which reads like a review (and probably is a review), there are many complex issues addressed concerning the structure of compounds interacting with the receptor. This is difficult to grasp if there is no graphical scheme that helps the reader to understand what we are talking about. But there is no such figure here at all. For example, what is E2, what is E2CCH? How does the ER binding pocket look like, how do the ligands with differential activities look? This has all been described and published in the past, but it would still help here to get something like a "graphical abstract".
Next, we enter the "data zone", where the author still very suddenly shifts over to experimental results - out of the blue. There is still no materials & methods section, not even rudimentary; please add this and I will be satisfied. So it remains unclear (to me) are these data based on purely biochemical assays, are there cell-based assays involved? Pleeeeeease clarify.
There is a lot of talk about chemically modified ligands/compounds binding to ER with a different mode, or activity. I would find it essential to introduce chemical structure of these compounds in a figure. This figure is missing. It should be coordinated with table 1 which summarizes the modifications.
Section 4.2: this is where we enter experimental data, and it needs to be clarified: end of the review section, beginning of the experimental section.
What are these "preliminary ERalpha binding assays? Please explain. How are they performed, helpful would also be to put references in there that explain the assay format from existing publications.
Section 4.2.1.: many chemical modifications are manetioned? What is their nature, where were they synthesized, who characterized them, who used them in binding assays? What is the binding assay used? How is it quantitated? And WHO did the experiments? Arent there any co-authors who actually did the work?
Section 4.2.2.: to begin with, it would be good if terms and abbreviations used here are explained. What is RBA? (this applies also to other sections). Same as with previous section, which assays were used, where are they descibed in detail, if you dont want to describe them here? But they cannot stay in the dark like they are now.
This is not an informative manuscript, since it appears isolated and disconnected from the previous body of work of the same author, and his previous research publications. That shouldnt be too difficult to change.
Figure 2 has significantly grown in size; but I still dont really understand what it all means, because its not properly explained in any details. The figure legends are still not satisfactory. For a large and complicated figure like this, you need more than 2 lines of explanation.
This article is difficult to appreciate for the reader whos not very deeply into the subject matter.
Comments on the Quality of English Language
it hasnt improved much. There are still many quality issues... some are mentioned in the review above. There are still grammatical errors, interpunctation issues, and "little things" like missing spaces etc. that may be fixed in the editorial process; but its going to be a lot of work.
There are also issues with the structure of sentences. I think they are often convoluted and difficult to understand; a more concise language and fewer fill words would also help. A native English speaker would most definitely help here.
Author Response
Reviewer stresses two points of major importance to be taken into account .
- The "hybrid" nature of the manuscript : partly review, partly original experimentation.
- Gramar issues of the manuscrpt , concering constructions of sentences and typing ( lack of commas, espaces between words).
I comment them before to adress specific remarcks concering our original work.
I totally understand the unappropriate hybrid aspect , even it was not pointed by the 3 other reviewers, justifying my previous attitude. Hence, I separated the the introduction ( review, sections 1-3) from the experimental work including its " materials and methods" aspect ( origin of compounds and experimental procedures ) before the presentation of our results ( Sction 4) . The last sentence of section 3 plays a link role with second part of the publication . This is easily detectable ( yellow color ; a procedure used in the whole manuscipt to identify changesi ). A text management without total change of the MDPI form.
Typing and gramar aspcts seem to me of minor importance ( lower amounts yellow color ) since the text was already corrected by MDPI , as could be see by comparison with PDFs ( corrections that I appreiated !). I anticipate that MDPI will similarilly operate in this second editoral round.
Specific remarks concerning the oginal aspect of the manuscript was systematically adressed, even some queries were unjustified , apparently generated by to lack of knowlege in the ERa thematic (E2: symbol of estradiol; - ECCH and related formulas: nature of a substituent of Hydroben in 11b position , RBA: Relative Binding Affinity ...). This jargon iwidely used eliminates its excessve explanation; idem for formula drawings of ligands commercial origins as well as fluorinated compouds of which origin and synthesis were notified by references. A chart of drawing of investigated ligands appears also ot necessary in view of their well known structures reprodueced eventually in publications. Collaborators who performed experiments long time ago have been notified in the reference of their work ; they could not be included as authors of the present publication since they are totally ignorant of the present status of which the publication is devoted
Requirement of drawing relevant to the ligand binding pocket has been taken into account by stressing the prominent importance of publication 28 ( L127) that reportrs a large set of figures including a mulitude of ligands insertions.
" Preliminary ERa binding assays " was remplaced by complementary assays deriving fom our investigation that may orient future sophisticated structural investigations evoked in our concludions remarks, an attitude proposed by another inestigator ( ref. 40)
Finally, I stresss the importance of Fig 2 of wich understanding looks to be difficult for the reviewer , but not for the 3 other reviewers. Its large size is required since its concerns the transcriptional a activity of all SERMs ( fluorinated or not) either alone or in the resence of control transcrtions inducer ( E2 for ERE , left; tamoxifen for AP1, right) to assesss possibilities of antagonism or synergy. An topic explained in the global view of our investigaitons ' (4.3.1.1 ; L209-215) to justify th originality of our experimental approch . A procedure aso used in the sssements of fuorinated SERDs on AP-1 transcription Fig 3).
Finally , I wish to mention that I failed to provide a tructurate th presentation of the manuscript , considering that MDPI will do it again. Moreover The editor manager resquest the manuscrpt for end of this year.
Dear Reviewer, I Hope that you appreciated my crrections that I did within less than a week .
