Title: Mechanism of NOTCH2 in Promoting Intrahepatic Bile Duct Development
Version: 1 Date: 23 May 2025
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
1. Lack of Novelty and Mechanistic Depth
The role of Notch2 in intrahepatic bile duct (IHBD) development is already well-documented.
The manuscript reiterates previous knowledge without providing significant new mechanistic
insights (e.g., downstream targets of Notch2 or how Notch2 precisely influences hepatoblast
differentiation beyond CK19 expression).
While the estradiol angle is mentioned, no molecular explanation is provided for estradiol
upregulating Notch2 or its ligands, limiting the translational impact.
2. Incomplete Characterization of the Model
The JAG1+/dDSL mutant mouse model, mentioned as a future direction, is not used in the
current study. Since this is the clinically relevant model of ALGS, excluding it undermines the
clinical relevance.
There is no evidence of direct Notch2 upregulation in ALGS models or human ALGS tissues,
which weakens the central hypothesis of Notch2 compensating for JAG1 mutations.
3. Over-reliance on In Vitro Data
The core findings seem primarily derived from in vitro hepatoblast cultures treated with estradiol
and siRNA. These models oversimplify in vivo bile duct development.
In vivo experiments are only superficially described, lacking robust quantitative data (e.g., bile
duct counts, morphometric analysis, liver histology).
4. Overstated Conclusions
The authors claim that Notch2 may "explain the clinical heterogeneity" in ALGS. This is
speculative and not supported by patient data or genotype-phenotype correlation analysis.
Statements like “compensatory activation of Notch2 with JAG2 or DLL1 may improve bile duct
development” are not experimentally tested, yet presented as near-certainties.
5. Language and grammar need polishing. Sentences are verbose and occasionally redundant
(e.g., repeating the definition of ALGS multiple times).
Title: Mechanism of NOTCH2 in Promoting Intrahepatic Bile Duct Development
Version: 1 Date: 23 May 2025
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
1. Lack of Novelty and Mechanistic Depth
The role of Notch2 in intrahepatic bile duct (IHBD) development is already well-documented.
The manuscript reiterates previous knowledge without providing significant new mechanistic
insights (e.g., downstream targets of Notch2 or how Notch2 precisely influences hepatoblast
differentiation beyond CK19 expression).
While the estradiol angle is mentioned, no molecular explanation is provided for estradiol
upregulating Notch2 or its ligands, limiting the translational impact.
2. Incomplete Characterization of the Model
The JAG1+/dDSL mutant mouse model, mentioned as a future direction, is not used in the
current study. Since this is the clinically relevant model of ALGS, excluding it undermines the
clinical relevance.
There is no evidence of direct Notch2 upregulation in ALGS models or human ALGS tissues,
which weakens the central hypothesis of Notch2 compensating for JAG1 mutations.
3. Over-reliance on In Vitro Data
The core findings seem primarily derived from in vitro hepatoblast cultures treated with estradiol
and siRNA. These models oversimplify in vivo bile duct development.
In vivo experiments are only superficially described, lacking robust quantitative data (e.g., bile
duct counts, morphometric analysis, liver histology).
4. Overstated Conclusions
The authors claim that Notch2 may "explain the clinical heterogeneity" in ALGS. This is
speculative and not supported by patient data or genotype-phenotype correlation analysis.
Statements like “compensatory activation of Notch2 with JAG2 or DLL1 may improve bile duct
development” are not experimentally tested, yet presented as near-certainties.
5. Language and grammar need polishing. Sentences are verbose and occasionally redundant
(e.g., repeating the definition of ALGS multiple times).
Please review this paper.
user
Title: Mechanism of NOTCH2 in Promoting Intrahepatic Bile Duct Development
Version: 1 Date: 23 May 2025
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
1. Lack of Novelty and Mechanistic Depth
The role of Notch2 in intrahepatic bile duct (IHBD) development is already well-documented.
The manuscript reiterates previous knowledge without providing significant new mechanistic
insights (e.g., downstream targets of Notch2 or how Notch2 precisely influences hepatoblast
differentiation beyond CK19 expression).
While the estradiol angle is mentioned, no molecular explanation is provided for estradiol
upregulating Notch2 or its ligands, limiting the translational impact.
2. Incomplete Characterization of the Model
The JAG1+/dDSL mutant mouse model, mentioned as a future direction, is not used in the
current study. Since this is the clinically relevant model of ALGS, excluding it undermines the
clinical relevance.
There is no evidence of direct Notch2 upregulation in ALGS models or human ALGS tissues,
which weakens the central hypothesis of Notch2 compensating for JAG1 mutations.
3. Over-reliance on In Vitro Data
The core findings seem primarily derived from in vitro hepatoblast cultures treated with estradiol
and siRNA. These models oversimplify in vivo bile duct development.
In vivo experiments are only superficially described, lacking robust quantitative data (e.g., bile
duct counts, morphometric analysis, liver histology).
4. Overstated Conclusions
The authors claim that Notch2 may "explain the clinical heterogeneity" in ALGS. This is
speculative and not supported by patient data or genotype-phenotype correlation analysis.
Statements like “compensatory activation of Notch2 with JAG2 or DLL1 may improve bile duct
development” are not experimentally tested, yet presented as near-certainties.
5. Language and grammar need polishing. Sentences are verbose and occasionally redundant
(e.g., repeating the definition of ALGS multiple times).
assistant