Designing an In Vivo Preclinical Research Study
, David Dworaczyk, Julia Whitehorn, Ke Li, Cristiano de Oliveira Cardoso and Abdelmotagaly Elgalad *
Round 1
Reviewer 1 Report
The manuscript, titled Designing an In Vivo Preclinical Research Study, provides a brief overview of the critical steps before and during preclinical studies and offers a short guide for scientists involved in preclinical research.
Although the overview is very brief, it covers important steps and aspects of preclinical research. This type of article can be important and useful for young scientists who lack experience in preclinical research, as it provides them with general guidelines and important checkpoints.
My objection relates to the overgenerality and lack of concrete examples and situations. However, considering that the authors had a word limit in mind, and I assume that they intended this review to be only a brief guide for future generations of investigators in preclinical research, I understand why they wrote the review in this style.
My only suggestion would be to absolutely include a section on ethical concerns that would cover the general ethical guidelines important for animal handling, surgical procedures, animal welfare, etc. I think that the data mentioned in the subsection on animal care is not sufficient and that this aspect should be expanded in a separate subsection.
Author Response
We have added this to the manuscript on lines 130-156.
Reviewer 2 Report
The review entitled 'Designing an In Vivo Preclinical Research Study' discusses about the three phases of effectively designing a preclinical research protocol: the research, preprocedural planning, and experimental phases. Here are my comments:
- Please discuss about statistics in details, i.e. power analysis, null hypothesis, parameteric vs non-parametric tests according to study design. This is of paramount importance for correct study design.
-Euthanasia and Necropsy in Fig 1 should be replaced by Euthanasia and 'Molecular/Histopathological Analyses'.
-Fig. 1 should also incorporate in vitro and in vivo under proof-of-concept. Please also discuss it in the text in detail as both are important for proof-of concept.
-Choice of euthanasia needs to be discussed according to study design, e.g. inhalational vs i.p. Decapitation may be required for certain tissues, e.g. brain.
-Modify this sentence 'produce the desired resultsand what evidence needs to be collected to support such an output'
-Authors write in table 'Rodents Adv:Drug therapy, skin wound healing, stroke'; however almost all preclinical studies are carried out in mice. They must include 'availability of reagents and knock-out/transgenic mice' in advantages. Also include other diseases such as asthma, multiple sclerosis, psoriasis, ect. This is very important.
minor
Author Response
- Please discuss about statistics in details, i.e. power analysis, null hypothesis, parameteric vs non-parametric tests according to study design. This is of paramount importance for correct study design.
RESPONSE: We have added this to the manuscript on lines 218-294.
-Euthanasia and Necropsy in Fig 1 should be replaced by Euthanasia and 'Molecular/Histopathological Analyses'.
RESPONSE: We made this change to Fig. 1.
-Fig. 1 should also incorporate in vitro and in vivo under proof-of-concept. Please also discuss it in the text in detail as both are important for proof-of concept.
RESPONSE: We aimed to describe the outline for an in vivo study, as described in the title. We believe that embarking on the design of an in vitro study is worthy of another review.
-Choice of euthanasia needs to be discussed according to study design, e.g. inhalational vs i.p. Decapitation may be required for certain tissues, e.g. brain.
RESPONSE: We have added this to the manuscript on lines 355-382.
-Modify this sentence 'produce the desired results and what evidence needs to be collected to support such an output'
RESPONSE: We have modified the sentence as suggested on lines 52-55.
-Authors write in table 'Rodents Adv:Drug therapy, skin wound healing, stroke'; however almost all preclinical studies are carried out in mice. They must include 'availability of reagents and knock-out/transgenic mice' in advantages. Also include other diseases such as asthma, multiple sclerosis, psoriasis, ect. This is very important.
RESPONSE: We have addressed this in Table 1.
Reviewer 3 Report
The manuscript entitled "Designing an In Vivo Preclinical Research Study" describes the main steps in the planning of a preclinical study. Mostly general descriptions were provided. The manuscript needs improvement.
Some examples from published research could be added.
The guidelines in preclinical research can be mentioned.
Especially 3R ethical rules in the use of experimental animals should be explained.
In the Table, the most used experimental animals mice and rats should be added. Also, monkeys are used in preclinical studies. The humanized mice could be mentioned, too.
Author Response
Some examples from published research could be added.
RESPONSE: We have cited some published examples throughout as suggested.
The guidelines in preclinical research can be mentioned.
RESPONSE: We have addressed this by adding mention of AVMA guidelines on lines 355-359.
Especially 3R ethical rules in the use of experimental animals should be explained.
RESPONSE: We have addressed this on lines 130-156.
In the Table, the most used experimental animals mice and rats should be added. Also, monkeys are used in preclinical studies. The humanized mice could be mentioned, too.
RESPONSE: We have addressed this in Table 1.
Reviewer 4 Report
In this review article, the authors reviewed and discussed the key points that preclinical researchers should consider when optimizing the design of an in vivo preclinical study to ensure that the findings will contribute to developing safe and effective treatments.
Comments
This is an interesting review article. The manuscript is well-writing. The reviewer has only some minor concerns as follows:
1. In line 50, “Figure. A phase-based diagram…” can be changed to “Figure 1. A phase-based diagram…”. Line 45 for “The Figure outlines…” can be changed to “The Figure 1 outlines…”.
2. In line 128, “Table. The significance….” can be changed to “Table 1. The significance….”. Line 115 for “The Table provides…” can be changed to “The Table 1 provides…”
3. In the Table, animal model for “Rodents” is too general and can be modified. It can be divided into “Rat” and “Mouse” for description.
Author Response
- In line 50, “Figure. A phase-based diagram…” can be changed to “Figure 1. A phase-based diagram…”. Line 45 for “The Figure outlines…” can be changed to “The Figure 1 outlines…”.
RESPONSE: We have addressed in the revised manuscript.
- In line 128, “Table. The significance….” can be changed to “Table 1. The significance….”. Line 115 for “The Table provides…” can be changed to “The Table 1 provides…”
RESPONSE: We have addressed this in the revised manuscript.
- In the Table, animal model for “Rodents” is too general and can be modified. It can be divided into “Rat” and “Mouse” for description.
RESPONSE: We have expanded the description of the best uses for rodents. However, we will keep them as a single category due to their significant overlap. Mice are the most common preclinical research animal model, with rats being their most similar alternative. We had initially divided them into two different categories but found ourselves with almost two identical rows.
Round 2
Reviewer 2 Report
Authors have addressed my concerns, no further comments
minor
