Review for 10.1186/s12876-024-03174-2 (2024)

Title: miR-541 is associated with the prognosis of liver cirrhosis and directly targets JAG2 to
inhibit the activation of hepatic stellate cells
Version: 0 Date: 19 Oct 2023
Reviewer's report:
Comments to authors
The study investigates the role of miR-541 in liver cirrhosis, an area with unclear pathogenesis
and a need for non-invasive prognostic markers. The research reveals downregulation of
miR-541 in cirrhotic tissues and sera, correlating with disease severity and clinical outcomes.
Functional assays demonstrate miR-541's impact on hepatic stellate cell activation. JAG2, a
Notch signaling ligand, is identified as a direct target of miR-541. The study suggests miR-541
as a potential non-invasive biomarker and therapeutic target for liver cirrhosis.
Liver cirrhosis remains a challenging clinical condition with a complex and incompletely
understood pathogenesis. While current prognostic markers have limitations, identifying novel
non-invasive predictors of prognosis in cirrhotic patients is of paramount importance. The study
at hand focuses on miR-541, previously recognized as a tumor suppressor in hepatocellular
carcinoma, yet its role in liver cirrhosis remains unexplored. Through a combination of
molecular techniques, the authors aim to elucidate the potential significance of miR-541 in
this context. The findings could potentially introduce a paradigm shift in our approach to liver
cirrhosis prognosis and therapy.
This study provides valuable insights into the role of miR-541 in liver cirrhosis, shedding light on
its potential as a non-invasive prognostic biomarker and therapeutic target. The comprehensive
approach, combining real-time PCR, functional assays, and bioinformatics analysis, strengthens
the validity of the findings. However, further studies could benefit from exploring the clinical
applicability of miR-541 in larger patient cohorts and assessing its potential in combination
with existing prognostic markers. Additionally, mechanistic details underlying the interaction
between miR-541 and JAG2 in the context of Notch signaling could be elucidated for a more
comprehensive understanding. Overall, this work significantly advances our understanding
of liver cirrhosis pathogenesis and offers promising avenues for clinical intervention. My
comments are as follows:
Major Comments:
1. Clinical Correlation: While the study establishes an association between miR-541 expression
and liver cirrhosis prognosis, it would greatly enhance the clinical relevance if additional
covariates (such as age, gender, etiology of cirrhosis, etc.) were considered in the survival
analysis. This would help in evaluating the independent prognostic value of miR-541.
2. Functional Validation: The functional assays performed to evaluate the effects of miR-541 on
LX-2 cells are critical for supporting the mechanistic insights. However, it might be beneficial
to include additional functional assays or complementary experiments to further validate the
observed effects, potentially using alternative models or cell lines.
3. Limitations of the Study Design: The study primarily focuses on miR-541 as a potential
biomarker and therapeutic target for liver cirrhosis. While this is valuable, it might be beneficial
to acknowledge the limitations of the study, such as the relatively small sample size in cohort
1 and the single-center nature of the study. Addressing these limitations would strengthen the
generalizability of the findings.
4. Exploration of Potential Confounders: The study could benefit from a more comprehensive
analysis of potential confounding factors that may influence miR-541 expression, both in
liver tissues and sera. This could include factors like medication usage, comorbidities, or
lifestyle variables. Accounting for these factors could provide a more nuanced understanding
of miR-541's role.
5. Therapeutic Implications: While the study identifies miR-541 and JAG2 as potential
therapeutic targets, it would be valuable to discuss the feasibility and challenges associated
with targeting these molecules in a clinical setting. Additionally, considering potential off-target
effects or compensatory mechanisms is crucial for the development of effective therapies.
Overall, the study significantly advances our understanding of miR-541 in liver cirrhosis.
Addressing these major comments would further strengthen the scientific rigor and clinical
relevance of the findings.
Minor Comments:
# Sample Size and Power Calculation: Providing a brief rationale or referencing previous
studies to justify the sample sizes in both cohorts would enhance the robustness of the study.
Additionally, including a power calculation could validate that the sample size is adequate to
detect meaningful effects.
# Blinding and Randomization: It would be helpful to mention if any blinding or randomization
procedures were employed during sample processing, data collection, or analysis to minimize
potential biases.
# Quality Control Measures: Detailing any quality control steps taken during RNA extraction
and real-time PCR, such as assessing RNA integrity or using appropriate controls, would provide
confidence in the reliability of the results.
# Normalization Strategy: Elaborating on the choice of reference genes or normalization method
for real-time PCR data would be valuable, as this step is crucial for accurate quantification of
miRNA expression.
# Statistical Methods: Providing more specific details on the statistical tests used (e.g., exact
tests, assumptions made) and addressing any adjustments for multiple comparisons can enhance
the transparency and rigor of the statistical analysis.
# Ethical Considerations: While informed consent and ethical approval are mentioned, it might
be beneficial to briefly outline any additional ethical considerations, such as patient anonymity
and data protection measures.
# Supplementary Methods: If there are additional methods or procedures not mentioned in the
main text but detailed in the supplementary materials, briefly referring to them in the main text
could help ensure completeness.
Overall, the methodology appears thorough and well-executed. However, providing additional
information on certain aspects, as suggested, would further strengthen the scientific rigor of the
study.