Therapies Targeted at Non-Coding RNAs in Prevention and Limitation of Myocardial Infarction and Subsequent Cardiac Remodeling—Current Experience and Perspectives

Round 1

Reviewer 1 Report

In the manuscript “Therapies targeted at non-coding RNAs in prevention and limitation of myocardial infarction and subsequent cardiac remodeling – current experience and perspectives” Kowara M and colleagues summarize the recent findings of the possible therapeutic applications in the field of targeting and regulating ncRNAs that are involved in the process of atherosclerotic plaque development, myocardial necrosis and cardiac remodeling after myocardial infarction. Moreover, the authors highlighted the importanceof the outcomes in preclinical studies with therapies targeted at ncRNAs. However, the reviewer has a number of issues and concerns that need to be addressed.

The reviewer would suggest some major and minor revisions.

Major Revisions:

The reviewer has found a lot of missing information about preclinical studies with miRNAs that are well known to be crucial in those pathologies, just to mention a few: miR-21 (the first study using miRNA therapeutics in a CVD model and demonstrated the role of miR-21 in regulating cardiac fibrosis), miR-199a-3p, miR-590-3p, miR-378, miR-17-3p, or HRCR and circ-FOXO3 (which are circular RNAs)…..etc Is there any reason why those miRNAs were not included? Maybe the reviewer is missing something, and if this is the case, the authors should explain this, if not this is really annoying when the authors are writing a review and they are missing such amount of information.

The section where the authors explain the agents that target ncRNAs need to be clarify and explain that there are different types of ASOs depending on the 2’sugar and backbone modifications: 2’-O-methoxyethyl (2’MOE), 2’-O-methyl (2’OMe), 2’-fluoro (2’F) and locked nucleic acid (LNA). Moreover, Table 1 should be revised, make it more clear. There are different kind of dots/squares that does not make any sense.

Redo Table 2. From the point of view of the reviewer need to be split it in two different tables and remove the red (miRNAs) and blue (lncRNAs) part of the table and put it without color, it will make it more clear. The legend figure is too long and difficult to understand. If there is no limit space, the authors may want to add a section explaining with more details Table2. The reviewer suggests to put another column with the reference/es.

In the Figure 2, the authors are missing a lot of information/papers from the miRNAs that they put as examples. I am not sure if it is only one example of each therapeutic agent that the authors wanted to point it out (so if this is the reason they have to state clear and explain the reason why). However, it might be confusing if they only put one, because for example in the case of miR-33 there is some controversy with the different studies that have been conducted. Which is the reason why the authors did not put all the different studies? I have just check the literature really quickly, and these are some articles that are missing:doi.org/10.1161/ATVBAHA.112.300639; doi.org/10.1161/ATVBAHA.113.301732; doi.org/10.1093/jmcb/mjz015;doi: 10.1097/SLA.0000000000001466;doi.org/10.1016/j.atherosclerosis.2017.04.011

 

At section 4, remove the example of the inhibitor of PCSK9, it is not targeting directly at ncRNA and may be confusing. Also in this section, the authors may want to add some information about HDLs or rHDLs that can be used as transporters of miRNAs or siRNAs. Although there are not many articles it might be interesting to add a little explanation as a possible future way to use HDL/rHDL as possible therapy tools. Here you have a couple of examples that use this technique 10.1016/j.jconrel.2019.07.007; 10.1016/j.jconrel.2018.05.041

 

Minor Revisions:

Double inverted commas in the introduction part should be check. As well in line 88.

Line 52 from the Figure 1, number 1 should be A, and number 2 should be B, right?, if not, the authors should explain what are these numbers from the figure.

Line 69, misspelling error in pathophysiological.

Line 117, misspelling error in lipoprotein.

Line 119, misspelling error in plaques.

Line 124, misspelling error in fragmentation

Line 138, missing a reference.

Line 133, misspelling error in stress.

Line 135, misspelling error in the sentence, my ncRNA? Should be by?

Line 168, there is no need to put the example of miR-155^-/-ApoE^-/-unless you add the reference, because there are dozens of papers that you could add there, so from the reviewer point of view, you should remove the example. The reviewer does not understand the point of the sentence that follow the miR-155^-/-ApoE^-/-, may the authors clarify it (MALAT and LincRNA-p21)

Line 273, should be miR-24

Line 294, without micro-RNA, control of miR-30.

Author Response

The manuscript describes very well the background of cardiovascular diseases and the implication of non-coding RNAs (miRNAs, circRNAs and lncRNAs) in the development and evolution of the processes associated to these human pathologies.

Major comments:

1. Line 37: I would like to know why authors consider miRs the most essential ncRNAs within short non-coding RNAs. Maybe they meant “currently better known” … Please, justify this affirmation. RESPONSE: I have changed to „currenlty best known group”.
   
   

2. Figure 1. The figure is called “Mechanism of action of miRNA and lncRNA”, however, the figure 1A) represents one of the pre-processing mechanisms of miRNAs and Figure 1B) represents a not very clear lncRNA structure, several lncRNAs functions (not all included) and the binding mechanism in a couple of the functions. This figure is very confused and does not clarify the text included in the manuscript. I recommend focussing the figure in one of the aspects (i.e. binding mechanisms of miRNAs and lncRNAs, or general functions of miRNAs and lncRNAs) in a clear manner. Recent recommended references for similar figures and/or tables: DOI: 10.3390/ijms21010302; DOI: 10.3390/ncrna7010017. RESPONSE: The Figure 1 has been revised according to the Reviever’s suggestions. Currently it concentrates upon principal mechanisms of action on both miRNA and lncRNA. The lncRNA shape has also been modified.
   
   

1. Line 57- 61. I do not consider as essential to explain in this kind of review the nomenclature of microRNAs and lncRNAs. I recommend eliminating these sentences. In fact, it does not have any connection with the following sentence in the paragraph. RESPONSE: the fragment about nomenclature has been removed, according to the suggestions.
   
   
2. Line 69-78. Please, rewrite this section. It is very difficult to understand the text included. RESPONSE: this section has been rewrited.
   
   
3. Table 1. There is not a clear distinction between different categories and subcategories within the concepts included in Inhibition and activation (circles, squares, black and white, etc).
   RESPONSE: The Table 1 has been corrected. I hope now it’s more clear and compatible with the text.
   
   
4. Line 157: I do not understand what authors mean when they write “virtually impossible”. I guess they tried to emphasize “the current difficulties of consecution” but this expression is redundant and not well expressed.
   RESPONSE: This phrase „virtually impossible” has been eliminated and the text has been corrected.
   
   
5. Line 159: Why did authors described as “exemplary studies” the representative works included in Figure 2? Please, justify this in the text if you consider that are better performed than other in the field. RESPONSE: The additional invesigations have been included into the Figure 2, according to the suggestions of the Reviewer 1. The phrase „exemplary studies” has been eliminated.
   
   
6. Figure 2. Several mistakes within the last line of boxes (C) of the figure (mice ApoE-/-, 18 weeks old): 1^st box below “therapeutic agent”, there is a “ ) ” missed at the end of the text; 3^rd box below “direct effect”, in the last line, the written text is “which makes” so it seems that there is some missing text. RESPONSE: The Figure 2 has been revised.
   
   
7. Please, rewrite the text between line 239-241, it is very difficult to understand. RESPONSE: Due to technical problems (extended HTML descriptions instead of normal reference numbers) I cannot precisely find the lines 239-241. However, I assume that these lines are located in 3.2 section, which has been widely reorganized.
   
   
   
8. Table 3. In row 1 (miR-144) column “Intervention”, include a reference “administration¹” and eliminate “(day 0, 1, 3 and later)”, because it is already described in the legend of the figure. Separate a little more the columns “Intervention” and “model”, because in the second row you can not distinguish the text belonging to each one.
   RESPONSE: I’m grateful for this comment, I’ll write a request to the Editor regarding Table 3
   
   
9. In the manuscript some references about human ongoing clinical trials related to ncRNAs are missed or the discussion about the stage of this kind of therapies applied to clinical practice. Authors only briefly describe the different types of therapies.
   RESPONSE: I’m grateful for this insightful comment. I’have managed to add a piece of information about ongoing clinical trial HERA, I have also mentioned completed trial with miravirsen.

Minor comments:

1. Line 15: write “circular RNA (circRNA)” instead of “circulatory RNA (circRNA)”.
2. Please, avoid the numerous contractions along the text, for example: it’s (line 27, 59, 80, 151, 153….), that’s (line 69), don’t (line 147), and some others.
3. Clarify the legends of the figures and tables. Please, organize them better and differentiate the points included because It is very difficult to understand.
4. There are some typos:
5. Line 117: write “lipoprotein” instead of “lipiprotein”
6. Line 119: “plaques” instead of “plauqes”
7. Line 132-133: duplication of the word “high laminar laminar” and correct the word “stress” (strss)
8. Line 137: “these networks have been excellently provided by…”
9. Line 273: Substitute “miR24” for “miR-24”
10. Please, review the text because there are some extra spaces, scores instead of commas, not clearly expressed concepts.

 

RESPONSE: The corrections according to the minor comments have been done

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript describes very well the background of cardiovascular diseases and the implication of non-coding RNAs (miRNAs, circRNAs and lncRNAs) in the development and evolution of the processes associated to these human pathologies.

Major comments:

1. Line 37: I would like to know why authors consider miRs the most essential ncRNAs within short non-coding RNAs. Maybe they meant “currently better known” … Please, justify this affirmation.

1. Figure 1. The figure is called “Mechanism of action of miRNA and lncRNA”, however, the figure 1A) represents one of the pre-processing mechanisms of miRNAs and Figure 1B) represents a not very clear lncRNA structure, several lncRNAs functions (not all included) and the binding mechanism in a couple of the functions. This figure is very confused and does not clarify the text included in the manuscript. I recommend focussing the figure in one of the aspects (i.e. binding mechanisms of miRNAs and lncRNAs, or general functions of miRNAs and lncRNAs) in a clear manner. Recent recommended references for similar figures and/or tables: DOI: 10.3390/ijms21010302; DOI: 10.3390/ncrna7010017.

3. Line 57- 61. I do not consider as essential to explain in this kind of review the nomenclature of microRNAs and lncRNAs. I recommend eliminating these sentences. In fact, it does not have any connection with the following sentence in the paragraph.
4. Line 69-78. Please, rewrite this section. It is very difficult to understand the text included.
5. Table 1. There is not a clear distinction between different categories and subcategories within the concepts included in Inhibition and activation (circles, squares, black and white, etc).
6. Line 157: I do not understand what authors mean when they write “virtually impossible”. I guess they tried to emphasize “the current difficulties of consecution” but this expression is redundant and not well expressed.
7. Line 159: Why did authors described as “exemplary studies” the representative works included in Figure 2? Please, justify this in the text if you consider that are better performed than other in the field.
8. Figure 2. Several mistakes within the last line of boxes (C) of the figure (mice ApoE-/-, 18 weeks old): 1^st box below “therapeutic agent”, there is a “ ) ” missed at the end of the text; 3^rd box below “direct effect”, in the last line, the written text is “which makes” so it seems that there is some missing text.
9. Please, rewrite the text between line 239-241, it is very difficult to understand.
10. Table 3. In row 1 (miR-144) column “Intervention”, include a reference “administration¹” and eliminate “(day 0, 1, 3 and later)”, because it is already described in the legend of the figure. Separate a little more the columns “Intervention” and “model”, because in the second row you can not distinguish the text belonging to each one.
11. In the manuscript some references about human ongoing clinical trials related to ncRNAs are missed or the discussion about the stage of this kind of therapies applied to clinical practice. Authors only briefly describe the different types of therapies.

Minor comments:

1. Line 15: write “circular RNA (circRNA)” instead of “circulatory RNA (circRNA)”.
2. Please, avoid the numerous contractions along the text, for example: it’s (line 27, 59, 80, 151, 153….), that’s (line 69), don’t (line 147), and some others.
3. Clarify the legends of the figures and tables. Please, organize them better and differentiate the points included because It is very difficult to understand.
4. There are some typos:
5. Line 117: write “lipoprotein” instead of “lipiprotein”
6. Line 119: “plaques” instead of “plauqes”
7. Line 132-133: duplication of the word “high laminar laminar” and correct the word “stress” (strss)
8. Line 137: “these networks have been excellently provided by…”
9. Line 273: Substitute “miR24” for “miR-24”
10. Please, review the text because there are some extra spaces, scores instead of commas, not clearly expressed concepts.

Author Response

The manuscript describes very well the background of cardiovascular diseases and the implication of non-coding RNAs (miRNAs, circRNAs and lncRNAs) in the development and evolution of the processes associated to these human pathologies.

Major comments:

1. Line 37: I would like to know why authors consider miRs the most essential ncRNAs within short non-coding RNAs. Maybe they meant “currently better known” … Please, justify this affirmation. RESPONSE: I have changed to „currenlty best known group”.
   
   

2. Figure 1. The figure is called “Mechanism of action of miRNA and lncRNA”, however, the figure 1A) represents one of the pre-processing mechanisms of miRNAs and Figure 1B) represents a not very clear lncRNA structure, several lncRNAs functions (not all included) and the binding mechanism in a couple of the functions. This figure is very confused and does not clarify the text included in the manuscript. I recommend focussing the figure in one of the aspects (i.e. binding mechanisms of miRNAs and lncRNAs, or general functions of miRNAs and lncRNAs) in a clear manner. Recent recommended references for similar figures and/or tables: DOI: 10.3390/ijms21010302; DOI: 10.3390/ncrna7010017. RESPONSE: The Figure 1 has been revised according to the Reviever’s suggestions. Currently it concentrates upon principal mechanisms of action on both miRNA and lncRNA. The lncRNA shape has also been modified.
   
   

1. Line 57- 61. I do not consider as essential to explain in this kind of review the nomenclature of microRNAs and lncRNAs. I recommend eliminating these sentences. In fact, it does not have any connection with the following sentence in the paragraph. RESPONSE: the fragment about nomenclature has been removed, according to the suggestions.
   
   
2. Line 69-78. Please, rewrite this section. It is very difficult to understand the text included. RESPONSE: this section has been rewrited.
   
   
3. Table 1. There is not a clear distinction between different categories and subcategories within the concepts included in Inhibition and activation (circles, squares, black and white, etc).
   RESPONSE: The Table 1 has been corrected. I hope now it’s more clear and compatible with the text.
   
   
4. Line 157: I do not understand what authors mean when they write “virtually impossible”. I guess they tried to emphasize “the current difficulties of consecution” but this expression is redundant and not well expressed.
   RESPONSE: This phrase „virtually impossible” has been eliminated and the text has been corrected.
   
   
5. Line 159: Why did authors described as “exemplary studies” the representative works included in Figure 2? Please, justify this in the text if you consider that are better performed than other in the field. RESPONSE: The additional invesigations have been included into the Figure 2, according to the suggestions of the Reviewer 1. The phrase „exemplary studies” has been eliminated.
   
   
6. Figure 2. Several mistakes within the last line of boxes (C) of the figure (mice ApoE-/-, 18 weeks old): 1^st box below “therapeutic agent”, there is a “ ) ” missed at the end of the text; 3^rd box below “direct effect”, in the last line, the written text is “which makes” so it seems that there is some missing text. RESPONSE: The Figure 2 has been revised.
   
   
7. Please, rewrite the text between line 239-241, it is very difficult to understand. RESPONSE: Due to technical problems (extended HTML descriptions instead of normal reference numbers) I cannot precisely find the lines 239-241. However, I assume that these lines are located in 3.2 section, which has been widely reorganized.
   
   
   
8. Table 3. In row 1 (miR-144) column “Intervention”, include a reference “administration¹” and eliminate “(day 0, 1, 3 and later)”, because it is already described in the legend of the figure. Separate a little more the columns “Intervention” and “model”, because in the second row you can not distinguish the text belonging to each one.
   RESPONSE: I’m grateful for this comment, I’ll write a request to the Editor regarding Table 3
   
   
9. In the manuscript some references about human ongoing clinical trials related to ncRNAs are missed or the discussion about the stage of this kind of therapies applied to clinical practice. Authors only briefly describe the different types of therapies.
   RESPONSE: I’m grateful for this insightful comment. I’have managed to add a piece of information about ongoing clinical trial HERA, I have also mentioned completed trial with miravirsen.

Minor comments:

1. Line 15: write “circular RNA (circRNA)” instead of “circulatory RNA (circRNA)”.
2. Please, avoid the numerous contractions along the text, for example: it’s (line 27, 59, 80, 151, 153….), that’s (line 69), don’t (line 147), and some others.
3. Clarify the legends of the figures and tables. Please, organize them better and differentiate the points included because It is very difficult to understand.
4. There are some typos:
5. Line 117: write “lipoprotein” instead of “lipiprotein”
6. Line 119: “plaques” instead of “plauqes”
7. Line 132-133: duplication of the word “high laminar laminar” and correct the word “stress” (strss)
8. Line 137: “these networks have been excellently provided by…”
9. Line 273: Substitute “miR24” for “miR-24”
10. Please, review the text because there are some extra spaces, scores instead of commas, not clearly expressed concepts.

 

RESPONSE: The corrections according to the minor comments have been done

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

In the revised version, the authors answered most of my concerns, but there are still some important points that need to be addressed

Major revisions:

Figure 2: references 41 and 42 did not check reverse cholesterol transplant (RCT), remove it from the direct effect in the figure.

Importantly, missing another paper about therapies using antimiR-33 and atherosclerosis mice model (doi: 10.1161/JAHA.112.003376). When the reviewer suggested to check the literature about those miRNAs that the authors put as examples in figure 2, the reviewer added few examples of some of the missing information. It is not the reviewer job to check for all of them, the authors have to do it. Check carefully if there is more missing information about the other examples of miRNAs in figure 2, please.

 

Still missing the justification about why the authors put in figure 2 those 4 miRNAs as example, are they the most important, the most well-known, the first ones to be describe???

Explain better why the conflicting results with antimiR-33 therapies in the different studies. The reviewer does not thing that are “little change in the design”, different therapeutics, different mice model, different approach to study atherosclerosis (regression vs progression), does not look like little changes. Please read more carefully those papers, it is important when you write a review that is not exactly your field to be more accurate.

Moreover, the reviewer has suggested to put the original references in table 2 and table 3. It is not acceptable that the authors use the references from different reviews, the authors have to put the originals (there is enough space to put it, like in Table 4)

Minor Revisions:

Is there any reason why the authors did not change the double inverted commas in the introduction? (As well in line 88) as I told in the first revision?

There are still some misspelling errors: in conseqence, barier,…..

Table 1, AAV-adenoviral vectors should be remove from the text

Missing a couple of “by” in Fasolo et al, and Pierce and Feinberg, should be by Fasolo, and by Pierce…Still missing a reference by Pierce and Feinberg

Author Response

Figure 2: references 41 and 42 did not check reverse cholesterol transplant (RCT), remove it from the direct effect in the figure.

Response: I have modified the Figure 2 and changed the tittle in the box to „DIRECT EFFECT  ON THE PLAQUE (INVESTIGATED IN A STUDY)”, then I have removed RCT from the mentioned boxes.

Importantly, missing another paper about therapies using antimiR-33 and atherosclerosis mice model (doi: 10.1161/JAHA.112.003376). When the reviewer suggested to check the literature about those miRNAs that the authors put as examples in figure 2, the reviewer added few examples of some of the missing information. It is not the reviewer job to check for all of them, the authors have to do it. Check carefully if there is more missing information about the other examples of miRNAs in figure 2, please.

Response: I’m grateful for this insightful comment. I have performed a literature review once again, in reference to miRNA and atherosclerosis. As a result, I have rearranged the Table 2, explained conflicting results and ambigous role of some miRNAs, described the role of miRNAs selected for the Figure 2 (miR-33, miR-145, miR-494, miR-495 and miR-98) and added information about other miRNA particles, which were studied with the use of their agonists or antagonists as therapeutic agents. Describing the role of miR-33, I have included suggested manuscript by Horie et al. which provided an evidence of atherogenic role of miR-33 (on double knock-out ApoE^-/- miR33^-/- mice). Moreover, I have also introduced an interesting study by Distel, which investigated the influence of anti-miR33 upon diabetic Reversa mice, a specially modified LDLr-/- mice in which hypercholesterolemia can be reversed by Mttp gene inactivation through polyinosinic polycytidylic RNA.  

Still missing the justification about why the authors put in figure 2 those 4 miRNAs as example, are they the most important, the most well-known, the first ones to be describe???

Response: I have explained, that these microRNA have been chosen because they regulate crucial elements of atherosclerotic plaque progression and were investigated by their agonists or antagonists (potential therapeutic agents). Moreover, miR-33 and miR-145 have been studied in more extensive way in the field of atherogenesis (which was also mentioned in the paper). I have also decided to add miR-98 to this figure, because it regulates LOX-1, an important factor in atheroslcerotic plaque development which contributes to macrophages conversion into foam cells.

Explain better why the conflicting results with antimiR-33 therapies in the different studies. The reviewer does not thing that are “little change in the design”, different therapeutics, different mice model, different approach to study atherosclerosis (regression vs progression), does not look like little changes. Please read more carefully those papers, it is important when you write a review that is not exactly your field to be more accurate.

Response: As I have mentioned in the response to the previous comment, I have rearranged the entire chapter refering to microRNA role and potential miRNA-targeted therapy. The information summarizing different results of studies investigating anti-miR-33 therapy has also been included.

Moreover, the reviewer has suggested to put the original references in table 2 and table 3. It is not acceptable that the authors use the references from different reviews, the authors have to put the originals (there is enough space to put it, like in Table 4)

Response: According to the Reviewer’s comment, I have rearranged Table 2 and 3 and included references from the original studies.  

Minor Revisions:

Is there any reason why the authors did not change the double inverted commas in the introduction? (As well in line 88) as I told in the first revision? – Response: I have checked the entire manuscript, now there is no double inverted commas.

There are still some misspelling errors: in conseqence, barier,…..Response: I have checked the entire manuscript, according to the comment.

Table 1, AAV-adenoviral vectors should be remove from the text Response: it has been removed.

Missing a couple of “by” in Fasolo et al, and Pierce and Feinberg, should be by Fasolo, and by Pierce…Still missing a reference by Pierce and Feinberg Response: I have corrected this error.

Reviewer 2 Report

The manuscript describes very well the background of cardiovascular diseases and the implication of non-coding RNAs (miRNAs, circRNAs and lncRNAs) in the development and evolution of the processes associated to these human pathologies. There is valuable information for the field related to the effects of targeting specific ncRNAs in preclinical studies.

The authors have completed the requests and added some valuable information and discussion about the pre-clinicial and ongoing clinical trials related to ncRNAs in cardiovascular diseases, improving considerably the quality of the manuscript. I am happy with the result of the changes and I would like to recommend this review for its publication, only with some minor corrections.

 

Just some minor comments:

1. Please, carefully check some typos and misspelling words in the text. For instance: “process” instead of “proces” (during the introduction and 3.1).
2. Section 3.1: “Myocardial infarction, in the vast majority of cases, is a …”
3. Table 3. There is a single asterisk (*) not correlating with anything in the table (indicates mRNAs which are targets for miRNAs…) and in fact not in the table.
4. Please, avoid the numerous contractions along the text, for example: it’s (still many of them there).
5. Clarify Figure 2 legend, is not readable. Please, organize them in a better way, for example: “A) target miR-33 [40]; B) target miR-33 [41]; C) target miR-33 [42]; D) target miR-145 [34] …”. Something consistent for all the sections.
6. Section 3.2.2.: “… inflammatory effect is caused by targeting [of] and degradation of…”; “Angiogenesis is a factor…”

Comments for author File: Comments.pdf

Author Response

The manuscript describes very well the background of cardiovascular diseases and the implication of non-coding RNAs (miRNAs, circRNAs and lncRNAs) in the development and evolution of the processes associated to these human pathologies. There is valuable information for the field related to the effects of targeting specific ncRNAs in preclinical studies.

The authors have completed the requests and added some valuable information and discussion about the pre-clinicial and ongoing clinical trials related to ncRNAs in cardiovascular diseases, improving considerably the quality of the manuscript. I am happy with the result of the changes and I would like to recommend this review for its publication, only with some minor corrections.

 Respose: I am very grateful for the Reviewer comment. I have corrected the mistakes indicated in the minor comments.

Just some minor comments:

1. Please, carefully check some typos and misspelling words in the text. For instance: “process” instead of “proces” (during the introduction and 3.1). Response: I have checked the entire manuscript once again.
2. Section 3.1: “Myocardial infarction, in the vast majority of cases, is a …Response: I have corrected this error.
3. Table 3. There is a single asterisk (*) not correlating with anything in the table (indicates mRNAs which are targets for miRNAs…) and in fact not in the table. Response: this error has been corrected.
4. Please, avoid the numerous contractions along the text, for example: it’s (still many of them there).Response: I have checked the entire text in order to eliminate excessive contractions.
5. Clarify Figure 2 legend, is not readable. Please, organize them in a better way, for example: “A) target miR-33 [40]; B) target miR-33 [41]; C) target miR-33 [42]; D) target miR-145 [34] …”. Something consistent for all the sections. Response: the Figure 2 legend has been corrected, according to the suggestion.
6. Section 3.2.2.: “… inflammatory effect is caused by targeting [of] and degradation of…”; “Angiogenesis is a factor…”Response: these errors have been corrected.

 

Round 3

Reviewer 1 Report

In the revised version, the authors answered most of my concerns, but there are still a couple of things that need to be addressed

 

Minor Revisions:

Put the same Table 2 that in the review-v2 that is more clear than the review-v3.

In Figure 2, the information in the TARGET column might be a little confuse. The reviewer thinks that with the information in the THERAPEUTIC AGENT and the DIRECT EFFECT columns are enough. For example, in number 1, if that study is using an anti-miRNA, it is clear that they are blocking the expression of that miRNA and some of the target genes that the authors check will be up-regulated. Please remove that information from TARGET column.   

Line 198- couple of didn’t, change it for did not

Line 205- misspelling error: Hovever

Author Response

Put the same Table 2 that in the review-v2 that is more clear than the review-v3.

Response: Although it was not possible to put the same version of Table 2 (Table 2 from review v2 has been rearranged and fulfilled with references from original studies) we have modified the Table 2 by elimination of some additional and unnecessary information. We hope that now Table 2 is more readable.

In Figure 2, the information in the TARGET column might be a little confuse. The reviewer thinks that with the information in the THERAPEUTIC AGENT and the DIRECT EFFECT columns are enough. For example, in number 1, if that study is using an anti-miRNA, it is clear that they are blocking the expression of that miRNA and some of the target genes that the authors check will be up-regulated. Please remove that information from TARGET column.   

RESPONSE: The TARGET column has been modified according to the suggestion in the comment.

Line 198- couple of didn’t, change it for did not - RESPONSE: Corrected.

Line 205- misspelling error: Hovever - RESPONSE: Corrected.