Molecular Biological Research on the Pathogenic Mechanism of Retinoblastoma
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript presents current knowledge on the molecular biology of retinoblastoma. The authors summarized recent studies about genetic mutations and epigenetic modifications involved in the development of retinoblastoma. Additionally, the authors provide some insights into targeted therapies for retinoblastoma. However, the manuscript suffers from confusing English language writing in several sentences, impeding its clarity. Very critically, the absence of several citations or inaccuracies in the citations undermines the scientific rigor and coherence of the manuscript.
Major concerns:
1. The storytelling in multiple sections is not logically focused, therefore it is challenging for the reviewer to follow the narrative thread and understand the progression of ideas. For instance, on line 36, the author initially mentions that "a small percentage of non-hereditary retinoblastomas are caused by…", but then introduces the two types of retinoblastomas: "RB is divided into hereditary and non-hereditary depending on the heredity." It would be clearer if the author first introduced the two types of RB, followed by providing genetic information about each type. Additionally, the authors discuss Claudin-1 in section 3.2.2; however, no relevant information about the gene is provided, rendering the paragraph difficult to comprehend.
2. The novelty of the current manuscript is limited. The genetics of retinoblastoma have been discussed in several recently published articles, and the current manuscript does not offer additional information beyond what is already available in the literature.
a. Marković, L., Bukovac, A., Varošanec, A.M. et al. Genetics in ophthalmology: molecular blueprints of retinoblastoma. Hum Genomics 17, 82 (2023).
b. Thériault, B. L., Dimaras, H., Gallie, B. L., & Corson, T. W. (2014). The genomic landscape of retinoblastoma: a review. Clinical & experimental ophthalmology, 42(1), 33–52.
3. The ‘Conclusion’ section of the manuscript is very weak. The conclusion section is not deep enough, to get adequate information for readers to understand the authors' implications. Besides highly conclusive words to summarize what the authors reviewed, the authors should also foresee the advantages and challenges of new treatment modalities, such as targeted therapies proposed in the manuscript.
4. As a review article, the authors should provide accurate and strong citations to support their statement. However, in the current manuscript, multiple references are missing, and some of the cited articles do not convey the information the author intends to express. The following are just small pieces of evaluation. For example, the citation is missing on lines 22-24. On line 33, the authors claimed that ‘RB is the first tumor caused by genetic change’, but this statement is not supported by any citation. RB is not the first identified cancer caused by genetic change, please provide the source of the statement. Additionally, on line 227, reference [51] is irrelevant to RB. On lines 48, 69, 79-81, 85-87, 141, 210 the citations are missing.
5. Sections 4.1, 4.2, and 4.3 seem unnecessary. The three sections are just basic definitions of epigenetic modifications. The authors should directly discuss the epigenetic characterizations of RB, without showing any unrelated information.
Minor concerns:
1. In this manuscript, the use of many punctuation marks is not appropriate. For instance, on line 281, ‘、’ should be ‘,’. In figure1 legend, ‘;’ use is wrong.
2. Figure2 spelling error, ‘Rtinoblastoma’ should be ‘Retinoblastoma’.
Comments on the Quality of English Language1. While the content of this manuscript is generally understandable, it requires intensive editing of the English language to enhance clarity, coherence, and overall readability. The following are just a few examples of areas needing improvement. For instance, on lines 33-35, the RB1 gene should be referred to as a "tumor suppressor gene" instead of "the suppressor one." Additionally, the sentence "The RB1 gene of genital cells…" on line 51 is overly long and hampers precise comprehension. Similarly, the sentence on lines 42-45 is excessively lengthy and difficult to comprehend. Furthermore, a grammatical error on line 134 undermines the clarity of the meaning. Lines 255-256 are also confusing and warrant clarification. The reviewer recommends that the authors collaborate with a writing coach or copy editor to enhance the flow and readability of the text.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors
Summary
This review article describes the molecular biology research of retinoblastoma with a focus on molecular biology. In contrast to other eye tumors such as uveal melanoma, retinoblastoma is due to a loss of the Rb1 gene in the retina that develops in children. Classical clinical treatment options are limited. Therefore, genomic studies, for example, are important to provide a deeper understanding of the pathogenesis of retinoblastoma and thus enable a more precise diagnosis and treatment of this disease. Epigenetic regulation also plays an important role in the development of retinoblastoma. They include non-coding RNAs, DNA methylation and histone modifications (epigenetics of retinoblastoma). This has implications for the diagnosis, treatment and prognosis of retinoblastoma (influencing epigenetic modifications). Clinical genetics (retinoblastoma gene RB1) is particularly relevant with regard to the different characteristics of hereditary and non-hereditary retinoblastoma. The pathogenesis of retinoblastoma involves abnormalities in several genes. The inactivation of two alleles of the tumor suppressor gene is necessary for the development of cancer ("two-hit theory") and leads to uncontrolled cell differentiation and tumor formation. The silencing of Rb1 leads (in the mouse model) to widespread growth of new retinal vessels with autophagy and dissolution of photoreceptors. The authors conclude, among other things, that biomarkers and gene expression characteristics have been identified that are associated with the prognosis of retinoblastoma and that there is currently (still) a lack of reliable prediction models for clinical diagnosis.
General Comments
This review is generally well written, easy to understand (no linguistic abnormalities). The introduction provides a good overview of the figures known from studies on retinoblastoma and the classic treatment approaches are limited (e.g. side effects after injection of medication into the vitreous). The structure of this manuscript is also easy to understand. In my opinion, the overview of the molecular biological pathogenesis of retinoblastoma and the insights into research into targeted therapies for retinoblastoma are well done. However, it would also be helpful to think outside the box in order to consider not only molecular biological aspects but also at least a few (important) cell biological approaches, which are naturally also related to molecular biology. Cell biology also concerns pathogenic mechanisms and can contribute to gaining knowledge. Otherwise, the title of the manuscript might have to be changed to "molecular genetic research" or something like that. However, molecular biology research is fine if the aspects mentioned above are also taken into account. Here, for example, apoptosis by calcium overload could also be mentioned (PMID: 12838338), which is relevant in tumor cell research. Clinically, the role of cytostatic drugs (e.g. etoposide) and the problem of resistance formation are also rarely addressed in this manuscript. There are numerous studies with well-established cell models (e.g. WERI-Rb1) that can lead to further insights and are mentioned here (only) with regard to the role of miRNA (as well as lncRNAs and circRNAs) in retinoblastoma (see e.g. PMID: 35409416 - protein profiling or PMID: 38339011 - etoposide insensitivity). I think the illustration in Fig. 1 (Genetics and Epigenetics of human Retinoblastoma) is well done. However, a few more illustrations (e.g. "two-hit theory" or pathological phenotypes) would have an additional positive effect.
Specific comments
1. Introduction
Which drugs are injected into the vitreous body? In addition, what about the side effects? What retinal damage is meant? Citations should be included.
3. Retinoblastoma genomics
Are there any contradictory studies on the "two-hit theory"? (See PMID: 20445799).
The same headings "Pathogenesis" in different chapters 4.1.1. 4.2.1. should be avoided.
5 Pathology of the RB
An illustration (table or graph) of the pathological phenotypes and the degree of tumor differentiation would be very useful in an overview (high versus low degree of tumor differentiation).
Minor
Fig. 2: Retinoblastoma (Typo).
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsWe thank the authors for their response. All the questions have been addressed, we have no further questions.
Comments on the Quality of English LanguageWe thank the authors for their response. All the questions have been addressed, we have no further questions.
