Pictilisib-Induced Resistance Is Mediated through FOXO1-Dependent Activation of Receptor Tyrosine Kinases in Mucinous Colorectal Adenocarcinoma Cells
Round 1
Reviewer 1 Report
Phosphatidylinositol 3-kinase (PI3K) targeting drugs are creating great interest as therapeutics for human cancers. PI3Ks belong to the lipid kinase family that are divided into three classes. Commonly, PI3KCA gene mutations occur in the kinase domain (e.g. H1047R) or in the regulatory domain (e.g. E542K, E545K). These mutations diminish inhibitory interactions leading to constitutive activation of PI3K and downstream effectors (e.g. AKT, mTOR, and S6 kinase). The hyperactivation of these signaling nodes regulate the PI3K/AKT axis towards the growth and survival of the cancer cell.
The title and content of the article represent a topic of real interest worldwide.
The subject of the study is topical with real interest for the future because the PI3K/AKT/mTOR axis is a target for cancers, the current use of small molecule inhibitors against this axis is limited by the development of drug-resistance in patients.Pictilisib treatment led to an increased accumulation of nuclear FOXO1 compared to vehicle treated LS174T cells. FOXO transcription factors are highly competitive transcriptional binders involved in determining the fate of target cells. FOXO1 involvement is a critical event in PI3K/AKT axis inhibition that mediates resistance in MCA cells.
The introduction of the article presents originality by proposing a topic with a huge academic potential.
The bibliographic data inserted along the article presents a qualitative chronology. The subject of the article represents a true scientific revolution in its field.
The material and methods section of the article presents a quantitative and qualitative exposition of the research plan, respectively a good reproducibility in order to develop other studies with this theme.
The results of the article present a logical and chronological exposition outlining qualitative aspects of the benefit. The figures and tables keep a specific chronology throughout their exposition, presenting qualitative aspects related to the subject of the article.
The topic of the article is a real interest for the future with major importance in this field. I consider it necessary to develop new studies on this subject and implement them on a population scale. The article presents an important research point with an optimal linguistic exposition, having an exponential potential for the future. This present article is written in a clear and concise manner.
The article presents originality, with an optimal literary exposition, representing a topic of real interest for the future with objective results at the research level. The article represents a launching platform in its field and from the point of view of the characteristics it is included for publication.
Reviewer 2 Report
First of all, thank you so much for involving me in reviewing this manuscript.
A very interesting and topical topic given that today genetics and molecular biology are becoming more and more involved in modifying therapies making them personalised.
Complex but well-conducted and understandable statistical analysis with well-structured graphs and scientifically valid results.
Clear and easily understood English language with easy reading.
Adequate and recent bibliography with a number of valid references.
Clear and understandable tables and images.
For me there are no changes to be made and the manuscript can be published.
Reviewer 3 Report
The manuscript is an original paper focused on the study of drug resistance induced in mucinous colorectal adenocarcinoma cells (MCA) by Pictilisib – a small molecule acting as a PI3K-inhibitor.
The introduction is concise well focused.
The Materials and Methods section is presented in detail and is easily reproducible. It is based on methodology which answers adequately the posed questions underlaying the aim of the study.
The results are organized in six subsections illustrated by 6 informative figures. The authors prove that FOXO1 nuclear accumulation correlates with the acute resistance in MCA cells after pictilisib treatment. FOXO1 regulates RTK rebound activity in pictilisib drug-induced resistant cells. FOXO siRNA attenuates phospho-ERK1/2 and phospho-AKT levels in case pictilisib is used as a single agent. If the FOXO1 inhibitor (AS1842856) and pictilisib are applied together, combinational synergy increases efficacy through cell toxicity and apoptosis.
The discussion is balanced and based on the original results of the authors. It would sound more persuasive if the limitations of the study are outlined. Although this is an experimental in vitro investigation it opens new perspectives for eventual therapeutic combinatorial treatment of mucinous colorectal adenocarcinomas with FOXO1i/PI3Ki agents.
The references are correctly cited. The English language is fine.
