The AKT1E17K Allele Promotes Breast Cancer in Mice
, Luca Roberto 4, Carmela De Marco 1, Marzia Scarfò 5, Donatella Montanaro 6, Orlando Paciello 7, Serenella Papparella 7, Chiara Mignogna 2,8, Alfonso Baldi 6,9 and Giuseppe Viglietto 1,2,*Round 1
Reviewer 1 Report
The authors have investigated the role of gain of function mutation E17K in AKT1 in driving mammary tumors using MMTV-CRE;R26-AKT1E17K mice models. They observed spontaneous mammary tumors in mice expressing AKT1E17 as compared to WT. Immunohistochemical characterization suggested basal like marker expression as they were PR-/HER2-/ER +, basal-like and CK8-/CK10-/CK5+ /CK14+. Further, GSEA of tumors expressing mutant AKT1E17K presented activation of the GSK3/cyclin D1 pathway in the mammary epithelium and clustering with the human basal-like tumors. They conclude that AKT1E17K is a bona fide oncogene that is initiates tumors at high efficiency in murine mammary epithelium.
Overall, it's an interesting research topic, exploring subtype formation from AKT mutant in breast cancer. Some concerns are as follows:
Figure 1C and 2A, please includes images with better resolution for both 20X and 40X magnification. ER expression is not clearly visible in the current figure.
Does the differences in dysplasia or proliferative index varied with or depend on parity of mice?
What percentage of tumors derived from AKTE17K were ER positive?
Can you include bar diagrams to depict IHC data in terms of percentage among low grade and high grade tumors.
Given the heterogenity in differentiation in tumors from AKTE17K, does the low differentiated tumors (that were not included in the analysis) express the markers or gene signatures derived from Gene array analysis or other basal like markers?
For some reason the supplementary files were not properly opening. Please make sure the correct version/format of files were uploaded.
Minor:
Please review the manuscript for spelling and space errors to maintain uniformity.
In methods, mice euthanasia was performed by inhaling carbon monoxide. Please make sure you follow the recommended guidelines for euthanasia, as Carbon dioxide is the most commonly used method for this purpose. Please correct if that was wrongly written.
Author Response
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Author Response File: 
Author Response.docx
Reviewer 2 Report
The aim of the present study was to evaluate the role of AKT1E17K in the transformation of breast epithelial cells in vivo. Two suggestions is as following:
In the Introduction, the authors mention that "Mutant AKT1E17K protein shows increased affinity for PI(4,5)P2, which allows its recruitment to plasma membrane and constitutive activation (15, 18)." It is better to evaluate PI(4,5)P2 binding affinity in the animal model.
Materials and methods: some more sentences to explain "mouse mammary tumor virus (MMTV)" is necessary (better).
Author Response
Please see the attachment
Author Response File: Author Response.docx
