Peer review of: Chronic activation of endothelial MAPK disrupts hematopoiesis via NFKB dependent (2020)


 
 

REVIEWERS' COMMENTS: 

Reviewer #1 (Remarks to the Author): 

With a great enthusiasm I read the revised manuscript and the response to reviewers’ critiques of 
Ramalingam et al. While, undoubtedly, the authors made a serious attempt to revise the manuscript 
according to the reviewers suggestions, I am disappointed to see that the major concerns of reviewer 
# 1 have not been appropriately addressed with new data, as requested ( see below). 

1. It still remains unclear if the justifications provided (such as “useful tool for scientific community” 
and “SCGF as a novel factor that enhances vascular and hematopoietic regeneration”) regarding the 
novelty of their study is convincing enough to publish in Nature communications. 

2. Reviewer # 1 requested the authors to provide data on distinct HSPC subsets; LT-HSC, ST-HSC, 
MPP2, MPP3 & MPP4, as these would be critical in understanding the physiological consequences of 
MAPK signaling defects in HSCs. However, the authors provided data following a HSC 
immunophenotyping strategy that is not accepted or followed by the field. Unfortunately, the 
immunophenotyping scheme (HPC-1, HPC-2 & MPP) presented by the authors is not consistent and 
contradicts with the HSC immunophenotyping strategies established by the pioneering labs, including 
Passegue lab ( Pietras, Cell Stem Cell, 2015; Yamashita, Cell Stem Cell, 2019) and Trumpp Lab 
(Wilson, Cell, 2008). For example, the authors show data on MPP without including Flt3 in their 
panels. This is unacceptable, as the “classical” MPPs established by the Weissman Lab are 
CD34+Flt3+LSK. Moreover, the authors did not provide data on relative numbers of any of these 
subsets. Finally, the rigor of the new data on HSPC is questionable, as they have shown data based on 
only n=5 mice. 

3. The rationale provided by the authors regarding Hypoxia is not at all convincing. Simply relying on 
the observations from some other Laboratory (of Tsvee Lapidot) is unacceptable, because, as we all 
know each experimental setting is different and data obtained from different mouse models can be 
completely different. The authors should have put some efforts in describing the downstream 
consequences of ROS in their model. 

4. Reviewer # 1 requested the authors to provide data on myeloid progenitors with a more refined 
immunophenotyping strategy (based on the strong published reports). Instead of showing these data 
with new experiments, the authors justify their immunophenotyping scheme by citing others (who still 
follow a controversial immunophenotyping scheme for defining myeloid progenitors), even though 
there are enough evidences to prove that this approach is incorrect. I do not believe that they 
provided a scientific justification for not showing the requested data. 

Reviewer #2 (Remarks to the Author): 

I commend the authors for the additional detailed analyses performed to address my concerns. Their 
revised manuscript has satisfactorily addressed the vast majority of my concerns. 

Reviewer #3 (Remarks to the Author): 

Endothelial MAPK activation disrupts hematopoiesis by including NFkappaB dependent inflammatory 
stress by Ramalingam et al. 
The authors addressed the concerns of this reviewer and almost all concerns of the other two 
reviewers sufficiently and convincingly, also with a large amount of novel data.