Escin Activates Canonical Wnt/β-Catenin Signaling Pathway by Facilitating the Proteasomal Degradation of Glycogen Synthase Kinase-3β in Cultured Human Dermal Papilla Cells

Round 1

Reviewer 1 Report

The manuscript investigated the stimulatory effects of escin on the canonical Wnt/β- catenin signaling pathway in cultured human dermal papilla cells(hDPCs). The data suggest that escin likely stimulates Wnt signaling through direct interactions with frizzled receptors.The experimental data as a whole are reasonable. But, there are still some elaboration are not sufficient.

After careful review, some detailed review comments are listed below:

1. Figure 3（c）lack of quantitative analysis.

2. Whether the activity of GSK-3β is related to phosphorylation is recommended to supplement or discuss phosphorylation of GSK-3β.

3. This paper studied escin activated Wnt/β-catenin signaling in HEK293 cells, but can it be applied to other tissue cells?

4. HEK293 cells and human dermal papilla cells are used in this paper. What is the relevance of this for the study of escin activated on Wnt/β-catenin signaling?

5. The main idea of the article is not specific, and further discussion on the pharmacological effects of escin.

The syntax is reasonable, some minor errors need to be fixed

Author Response

Cover letter

Dear Editor of CIMB.

In the 1st revision of our manuscript, i have answered or added data as you suggest below.
Thank you for your kind comment and suggestion.

 

Yours sincerely, Nae-Gyu Kang

 

 

<Revision>

 

The manuscript investigated the stimulatory effects of escin on the canonical Wnt/β- catenin signaling pathway in cultured human dermal papilla cells(hDPCs). The data suggest that escin likely stimulates Wnt signaling through direct interactions with frizzled receptors.The experimental data as a whole are reasonable. But, there are still some elaboration are not sufficient.

After careful review, some detailed review comments are listed below:

1. Figure 3（c）lack of quantitative analysis.
   - We added quantitative analysis data in Figure 3 (c).
2. Whether the activity of GSK-3β is related to phosphorylation is recommended to supplement or discuss phosphorylation of GSK-3β.
   - It was reported that the inhibitory phosphorylation on GSK3-β regulate the wnt/β-catenin signaling pathway, we described in discussion section.
3. This paper studied escin activated Wnt/β-catenin signaling in HEK293 cells, but can it be applied to other tissue cells?
   - TOPFlash activity of wnt/β-catenin HEK293 cell were reported in previous hair follicle studies. As reviewer mentioned, we cross checked the activation of β-catenin and Lef-dependent transcription in HEK293 TOPFLASH reporter cell line and primary dermal papilla cells.
4. HEK293 cells and human dermal papilla cells are used in this paper. What is the relevance of this for the study of escin activated on Wnt/β-catenin signaling?
   - We screened over 400 candidates chemicals which could activate β-catenin and Lef-dependent transcription signaling in HEK293 TOPFLASH reporter cell line. Among the chemicals, escin showed the best efficacy. After screening, we confirmed the β-catenin activation effect in primary human dermal papilla cells to improve male/female pattern hair loss.
5. The main idea of the article is not specific, and further discussion on the pharmacological effects of escin.
   - described new discussion sentences.

 

Author Response File: Author Response.DOCX

Reviewer 2 Report

The findings appear to be interesting. Major points that the authors need to address are as follows:

1. The molecular mechanism by which Escin stimulated Wnt/β-catenin signalling should be investigated in detail

2. The rationale for focussing only on escin is not clear. How this compound was selected?

3. A limited in vivo study will greatly increase the impact of the findings.

4. Can escin also activate other signalling pathways in addition to Wnt/β-catenin in human dermal papilla cells.

5. The manuscript should be carefully checked for typographical errors.

Moderate editing of English is required.

Author Response

Cover letter

Dear Editor of CIMB.

In the 1st revision of our manuscript, i have answered or added data as you suggest below.
Thank you for your kind comment and suggestion.

 

Yours sincerely, Nae-Gyu Kang

 

<Revision>

The findings appear to be interesting. Major points that the authors need to address are as follows:

1. The molecular mechanism by which Escin stimulated Wnt/β-catenin signalling should be investigated in detail
- More detailed mechanism should be elucidated especially focused on upstream signal interaction between escin and Frizzled receptors, LRPs.

2. The rationale for focussing only on escin is not clear. How this compound was selected?
   - We screened over 400 candidates chemicals which could activate β-catenin and Lef-dependent transcription signaling in HEK293 TOPFLASH reporter cell line. Among the chemicals, escin showed best efficacy. After screening, we confirmed the β-catenin activation effect in primary human dermal papilla cells to improve the alopecia areata, male/female pattern hair loss.
3. A limited in vivo study will greatly increase the impact of the findings.
   - Ex-vivo hair follicle culture and human in-vivo test of escin are now in progress. Roughly anagen elongative effect were observed in escin treated samples. Specific data will be shown in next paper.
4. Can escin also activate other signalling pathways in addition to Wnt/β-catenin in human dermal papilla cells.
   - We also confirmed not only wnt/ β-catenin signal induction, but several growth factors which elongate anagen and abrogate telogen phase were significantly induced and reduced. As, reviewer mentioned, further study will be shown in next papar.
5. The manuscript should be carefully checked for typographical errors.
   - We checked typographical errors.

 

 

Author Response File: Author Response.DOCX

Round 2

Reviewer 2 Report

The authors have addressed all my concerns.

The revised manuscript can be accepted now.