Immune Pathway and Gene Database (IMPAGT) Revealed the Immune Dysregulation Dynamics and Overactivation of the PI3K/Akt Pathway in Tumor Buddings of Cervical Cancer

Round 1

Reviewer 1 Report

The article titled "Immune Pathway and Gene Database (IMPAGT) Revealed the Immune Dysregulation Dynamics and Overactivation of the PI3K/Akt Pathway in Tumor Buddings of Cervical Cancer" is a well written and detailed report of the molecular findings in cervical carcinoma tumour budding.

As a Histopathologist working in the subspeciality field of gynaecology oncology we have not incorporated the reporting of tumour budding in our cervical cancer reports.

Previous studies however have demonstrated that tumour budding is  associated with overall inferior survival rates in cervical cancer.  The definition of tumour budding and high tumour budding does however vary between studies.

I am assuming the cervical carcinomas that were selected for the study are all HPV associated and not HPV-independent eg gastric-type endocervical adenocarcinomas.  Gastric-type adenocarcinomas of the cervix are Grade 3 tumours and often present with advanced stage disease.  A sentence to clarify this distinction may be of value to the reader.

 

 

Author Response

Dear Reviewer,

Thank you very much for your comment. We analyzed HPV genotype and the prevalence of HPV-negative tumors following your comment. The most common HPV type was 16 (52.4%). HPV was not detected in 3 patients (14.3%) with all adenocarcinoma. Among 3 HPV-negative cases, only one patient had HPV-independent gastric-type endocervical adenocarcinoma. Details of prevalence of HPV-type was added in Table 1.

Sincerely yours,

Yeseul Choi

Reviewer 2 Report

Choi et. al. investigated the tumor immune microenvironment of tumor buddings in cervical cancer. They performed RNA sequencing on patients' samples with high or low tumor budding. They generated Immune Pathway and Gene Database (IMPAGT) for immune profiling and found that PI3K/Akt/mTOR signaling pathway is overexpressed in the tumor immune microenvironment of cervical cancer with high tumor budding. The work is preliminary and validations are needed to draw some conclusions.

1-   21 cervical cancer samples (15 with high tumor budding, and 6 with low tumor budding) are a very small number of samples to draw a conclusion. More samples are necessary.

2-   The authors should perform IHC in the tumor tissues to confirm the expression of PI3K/Akt/mTOR.

 

 

Author Response

Dear Reviewer,

Please see the attachment.

I have also copied the Response here for your convenience as well. 

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1-   21 cervical cancer samples (15 with high tumor budding, and 6 with low tumor budding) are a very small number of samples to draw a conclusion. More samples are necessary.

Thank you very much for your comments. As you mentioned, 21 samples may be relatively a small number to elucidate a conclusion of a study. However, considering the difficulty and longer time period required to collect large number of clinical samples, 21 Cervical Cancer samples were the optimal amount we were able to analyze by then. Aware of its importance, however, we did have been continuously collected samples to complement current limitation in our next study. Since we were able to reach an interesting result that provided more specific direction of future study, we plan to conduct more constructed study with larger clinical samples as our follow-up research.

2-   The authors should perform IHC in the tumor tissues to confirm the expression of PI3K/Akt/mTOR.

We have not performed protein level analysis such as Western blotting or Immunohistochemistry (IHC) in current study. However, those analyses are scheduled in our follow-up study that is soon to be initiated with larger number of samples. There are few references that presented involvement and enrichment of PI3K/Akt signaling pathway in Cervical Cancer. Zhang et al, for example, discovered that both PI3K/Akt/mTOR gene and protein expression levels were positively correlated with the malignancy degree in Cervical Cancer tissues with qPCR and IHC; higher in Cervical Intraepithelial Neoplasia (CIN) patients and markedly higher in Cervical Cancer patients compared to that in healthy control group [1]. Other references also validated involvement of PI3K/Akt/mTOR signaling pathway in relation to progression of Cervical Cancer with protein level analysis via other biological markers [2,3].

References

1. Zhang, W.; Zhou, Q.; Wei, Y.; Da, M.; Zhang, C.; Zhong, J.; Liu, J.; Shen, J. The Exosome-Mediated PI3k/Akt/MTOR Signaling Pathway in Cervical Cancer. Int. J. Clin. Exp. Pathol. 2019, 12, 2474–2484.
2. Hu, R.; Wang, M.Q.; Niu, W.B.; Wang, Y.J.; Liu, Y.Y.; Liu, L.Y.; Wang, M.; Zhong, J.; You, H.Y.; Wu, X.H.; et al. SKA3 Promotes Cell Proliferation and Migration in Cervical Cancer by Activating the PI3K/Akt Signaling Pathway 11 Medical and Health Sciences 1112 Oncology and Carcinogenesis. Cancer Cell Int. 2018, 18, 1–16, doi:10.1186/s12935-018-0670-4.
3. Rong, L.; Li, H.; Li, Z.; Ouyang, J.; Ma, Y.; Song, F.; Chen, Y. FAM83A as a Potential Biological Marker Is Regulated by MiR-206 to Promote Cervical Cancer Progression Through PI3K/AKT/MTOR Pathway. Front. Med. 2020, 7, 1–11, doi:10.3389/fmed.2020.608441

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Sincerely yours,

Yeseul Choi 

Author Response File: Author Response.pdf