ITIH4 in Rheumatoid Arthritis Pathogenesis: Network Pharmacology and Molecular Docking Analysis Identify CXCR4 as a Potential Receptor
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
The research article titled "Integrating Network Pharmacology and Molecular Docking to Reveal the Mechanistic Role of Inter-alpha Trypsin Inhibitor Heavy Chain 4 (ITIH4) in Rheumatoid Arthritis Pathogenesis" investigates the potential role of ITIH4 in the development of Rheumatoid Arthritis (RA). Through the use of network pharmacology and molecular docking techniques, the study examines the interactions between ITIH4 and proteins associated with RA, with a particular focus on the chemokine signaling pathway. The findings suggest that ITIH4 may interact with CXCR4, a key chemokine receptor, potentially exacerbating RA by promoting inflammation via this pathway. The study proposes that targeting the ITIH4-CXCR4 interaction could offer a promising therapeutic strategy for managing RA.
However, a significant limitation of the paper is that it relies solely on computational methods, such as network pharmacology and molecular docking. While these methods provide valuable preliminary insights, they lack experimental validation. The findings must be confirmed through in vitro and in vivo studies to establish their biological relevance and potential clinical applications.
Moreover, the authors should carefully review the manuscript to address any typographical errors, labeling inconsistencies, and formatting issues to enhance the clarity and professionalism of the presentation.
Author Response
Comment: The research article titled "Integrating Network Pharmacology and Molecular Docking to Reveal the Mechanistic Role of Inter-alpha Trypsin Inhibitor Heavy Chain 4 (ITIH4) in Rheumatoid Arthritis Pathogenesis" investigates the potential role of ITIH4 in the development of Rheumatoid Arthritis (RA). Through the use of network pharmacology and molecular docking techniques, the study examines the interactions between ITIH4 and proteins associated with RA, with a particular focus on the chemokine signaling pathway. The findings suggest that ITIH4 may interact with CXCR4, a key chemokine receptor, potentially exacerbating RA by promoting inflammation via this pathway. The study proposes that targeting the ITIH4-CXCR4 interaction could offer a promising therapeutic strategy for managing RA.
However, a significant limitation of the paper is that it relies solely on computational methods, such as network pharmacology and molecular docking. While these methods provide valuable preliminary insights, they lack experimental validation. The findings must be confirmed through in vitro and in vivo studies to establish their biological relevance and potential clinical applications.
Response: Thank you very much for appreciating our work.
We also deeply appreciate your kind effort to review our manuscript and provide insightful suggestions; as per your kind suggestion, we validated our in-silico result through in-vitro studies in fibroblast-like synoviocytes (FLS) isolated from the synovium of RA patients (line number 155-211). Our data suggested a link between ITIH4 and CXCR4 expression, depicted in Figure 6 (line number 311-332), discussed in line number 411-417.
We have now revised our manuscript and highlighted the same in yellow color.
Comment: Moreover, the authors should carefully review the manuscript to address any typographical errors, labeling inconsistencies, and formatting issues to enhance the clarity and professionalism of the presentation.
Response: Thank you very much for your kind suggestion. We have improvised our manuscript according to your kind suggestion (Highlighted yellow in the revised manuscript). We now believe that our manuscript is improved and achieved standard of your esteemed journal “pathophysiology”.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors
The authors utilized network pharmacology and molecular docking to identify ITIH4's interacting proteins and their potential involvement in RA's molecular pathways. The approach is commendable and the computational findings are of interest.
However, the manuscript's findings are predicated on in silico analysis, which, while suggestive, require empirical substantiation. The identified molecules and pathways implicated in RA's pathogenesis through ITIH4 interaction lack the support of experimental validation. It is essential for the scientific community and for the advancement of the study's impact that these computational hypotheses are rigorously tested through appropriate biological assays. I recommend that the authors complement their work with either cell-based experiments or animal models to validate the interaction of ITIH4 with the identified proteins and their relevance to RA. This validation is critical to affirm the study's conclusions and to ensure that the proposed molecular mechanisms have a solid experimental foundation.
Additionally, the manuscript presents a hypothetical model in Figure 6, depicting ITIH4's mechanistic pathway via CXCR4 activation in RA synoviocytes. This model includes several key elements that are not discussed in the results. For the manuscript to meet the standards of evidence-based research, the authors should either provide the necessary experimental data to support the model or revise it to reflect only the pathways and molecules that have been empirically addressed within the study's scope.
Addressing these concerns will greatly enhance the manuscript's scientific rigor and its contribution to the field of rheumatology.
Comments on the Quality of English Language
The manuscript is generally well-written, with clear and concise language that effectively communicates the study's findings. The authors have made a commendable effort to present their research in a coherent and organized manner. The use of technical terms is appropriate, and the language is suitable for a scientific audience.
Author Response
Comment: The authors utilized network pharmacology and molecular docking to identify ITIH4's interacting proteins and their potential involvement in RA's molecular pathways. The approach is commendable and the computational findings are of interest.
However, the manuscript's findings are predicated on in silico analysis, which, while suggestive, require empirical substantiation. The identified molecules and pathways implicated in RA's pathogenesis through ITIH4 interaction lack the support of experimental validation. It is essential for the scientific community and for the advancement of the study's impact that these computational hypotheses are rigorously tested through appropriate biological assays. I recommend that the authors complement their work with either cell-based experiments or animal models to validate the interaction of ITIH4 with the identified proteins and their relevance to RA. This validation is critical to affirm the study's conclusions and to ensure that the proposed molecular mechanisms have a solid experimental foundation.
Response: Thank you very much for appreciating our work. We also thank and deeply appreciate for your effort to review our manuscript and to provide your insightful suggestion. Accordingly, we have undertaken in-vitro experiments to validate our in-silico findings. Specifically, we conducted studies using fibroblast-like synoviocytes (FLS) isolated from the synovial tissue of rheumatoid arthritis (RA) patients (see lines 155-211 in the revised manuscript). Our experimental data revealed a significant association between ITIH4 and CXCR4 expression, which is now illustrated in the updated Figure 6 (lines 311-332). Moreover, we have significantly expanded the result section to integrate this crucial experimental validation (lines 334-345), further supporting the proposed mechanistic pathway in updated Figure 7. We believe these additional data significantly strengthen our manuscript by providing a robust empirical foundation for our computational predictions.
Additionally, the manuscript presents a hypothetical model in Figure 6, depicting ITIH4's mechanistic pathway via CXCR4 activation in RA synoviocytes. This model includes several key elements that are not discussed in the results. For the manuscript to meet the standards of evidence-based research, the authors should either provide the necessary experimental data to support the model or revise it to reflect only the pathways and molecules that have been empirically addressed within the study's scope.
Addressing these concerns will greatly enhance the manuscript's scientific rigor and its contribution to the field of rheumatology.
Response: Thank you for your invaluable suggestion. In Figure 6 (Updated as Figure 7 in the revised manuscript), we have presented a hypothetical model depicting ITIH4's mechanistic pathway via CXCR4 activation in RA synoviocytes. As per your suggestion, now we have introduced in the result section a paragraph discussing the hypothetical model elaborating signaling pathways regulated by CXCR4 activation by ITIH4 in RA-FLS (lines 334-345).
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Comments and Suggestions for Authors
The revised manuscript has been substantially improved.
The incorporation of experimental data, particularly the association between ITIH4 and CXCR4 expression in FLS from RA patients, provides a solid empirical foundation for your computational predictions.
The discussion of the signaling pathways regulated by CXCR4 activation by ITIH4 in RA-FLS is a valuable addition that aligns with the empirical data presented.
Comments on the Quality of English Language
