Deciphering Glioblastoma: Fundamental and Novel Insights into the Biology and Therapeutic Strategies of Gliomas

Round 1

Reviewer 1 Report

The paper entitled “Deciphering Glioblastoma: A Deep Dive into the Biology and Therapeutic
Strategies of Gliomas” by Razvan Onciu and coworkers investigates in a very broad way the
epidemiological, biological and therapeutic characteristics of glioblastoma. In addition, to
examining the immunological context, this work analyzes what has been published on the
biochemical pathways involved in tumor progression and examines the different therapeutic
strategies based on the signaling pathways found altered in glioblastomas. Through the analysis of
published transcriptomic research, it highlights the variations in gene expression typical of this
tumor with a poor prognosis, evaluating the possible prognostic and therapeutic implications. The
role played by the microenvironment, not only immunological, and by epigenetic alterations is also
taken into consideration. We could therefore say that this review of literature data takes into
consideration all the salient biological aspects of glioblastoma.
General comments
The work as a whole is interesting, but many acronyms, especially gene names, are not explained
and this makes reading the manuscript difficult. Furthermore, not all readers, although interested in
the topic, know the genes involved and their biological role to understand the importance of their
mutations in the development and progression of glioblastoma. Therefore, I would recommend
making a table in which to insert all the genes whose mutation, or alteration of expression, etc. is
important in glioblastoma. This table should contain the name of the genes, their biological
function, and the relevant reference(s) reporting for the first time their involvement in glioblastoma.
Specific points
1. Lines 149/155. The authors could distinguish the three types of gliobastona by explaining
the molecular alterations that characterize them.
2. The meaning of what is written in lines 432/437 is unclear to me. Please, clarify
3. Speaking of metabolism, information on energy and lipid metabolism in glioblastoma
should be added.
4. The role of the nervous system in cancer regulation is increasingly appreciated. To this end,
the authors should include some sentences citing, for example, this interesting work
(Taylor KR, Barron T, Hui A, Spitzer A, Yalçin B, Ivec AE, Geraghty AC, Hartmann GG, Arzt M,
Gillespie SM, Kim YS, Maleki Jahan S, Zhang H, Shamardani K, Su M, Ni L, Du PP, Woo PJ,
Silva-Torres A, Venkatesh HS, Mancusi R, Ponnuswami A, Mulinyawe S, Keough MB, Chau I,
Aziz-Bose R, Tirosh I, Suvà ML, Monje M. Glioma synapses recruit mechanisms of adaptive
plasticity. Nature. 2023 Nov;623(7986):366-374. doi: 10.1038/s41586-023-06678-1.)
5. In the chapter dedicated to the microenvironment, the authors should include some
information regarding estrogens, as there are some interesting papers on the topic.
6. As regards treatments, the authors should also include some information on the possibility
of considering the therapeutic use of drugs used for psychiatric pathologies that have
shown therapeutic activity in glioblastoma with a view to a drug repositioning strategy.

I am not qualified to assess the quality of English in this paper

Author Response

We are thankful to Reviewer 1 for their appreciative remarks and constructive feedback on our manuscript.

1. Gene Names and Acronyms: We have included a detailed table summarizing most important genes involved in glioblastoma and their biological functions. This table is designed to aid readers in understanding the role of the most important genes in GBM

2. Molecular Alterations in Glioblastoma: We have elaborated on the molecular alterations characterizing the different types of glioblastoma by improving Figure 1

3. Clarification and Addition of Metabolic Information: We have clarified the ambiguous statements you highlighted and expanded our discussion on energy and lipid metabolism in glioblastoma

4. Role of the Nervous System: We have incorporated the work you have talked about in the importance of the CNS in GBM

5. Microenvironment and Estrogens: We have expanded our discussion on the tumor microenvironment to include the influence of estrogens- added estrogen receptors that are found in the GBM and their microenvironment, added the fact that they help in tumor growth and cellular proliferation and how targeting them can be a viable treatment option for GBM in the future.

6. Therapeutic Use of Psychiatric Drugs: In line with your suggestion, we have detailed the promising approach of drug repositioning, specifically highlighting the therapeutic potential of psychiatric drugs in glioblastoma treatment.

Reviewer 2 Report

Comments for author File: Comments.pdf

Author Response

We sincerely appreciate the constructive feedback and insightful suggestions provided by Reviewer 2 regarding our manuscript. Your comments have significantly contributed to enhancing the quality and depth of our review on glioblastoma biology and therapeutic strategies.

1. Abbreviations: We have meticulously reviewed and revised the abbreviations throughout the manuscript to ensure consistency and clarity, in line with your recommendation.

2. Glioma Complexity: Acknowledging the complexity of glioma subtypes and your suggestion to add a scheme, we have enhanced section 1.1 by adding more details to Figure 1

3. Mutations and Therapies Table: In response to your suggestion, we have included a comprehensive table outlining the key mutations in gliomas and corresponding therapies, both approved and under development. We have aimed to keep the information brief in the table since the manuscript is long, but if you feel like there should be more genes added we are happy to do so

4. Figure on Crosstalk in Gliomas: We have added a figure illustrating crosstalk in gliomas, Figure 3

5. Reference Update: We have updated our references, by incorporating a newer bibliography. However, we believe that even though some studies are older, they represent important studies when taking into account this field and therefore we believe they should be kept in the manuscript.

6. Therapeutic Strategies Detailing: We have expanded our discussion on therapeutic strategies, including: targeting estrogen receptors, the repositioning of antipsychotic drugs and ferroptosis. This enhancement aims to provide a more comprehensive overview of the innovative approaches being explored in glioblastoma treatment.

Your feedback has been invaluable in refining our manuscript, and we believe that by adressing the ammendments provided by your expertise we have enriched the quality of our manuscript.

Reviewer 3 Report

Comments for the manuscript by Onciu et al:

The manuscript is overwhelming providing a lot of information that is not in order and not linked, while there are parts that are repetitive and whole paragraphs that do not correspond to the given title.

There are sentences and used verbs that do not help with the understanding and the flow of the text. Such examples are:

-lines: 29-31, 149-151, 274-275

-verbs: believed (line38), signaling (line 40), manifest (line 52), precipitate (line 183-184)

Other comments that sound minor but collectively add to the difficulty of reading the text and sounds inaccurate:

-              Line 57-58: brain tumors are involved in the CNS tumors.

-              Line 62 and 84 are repetitions of the same info with different statistics.

-              What is the 2021 classification of WHO that is referred to in line 140? What is the immunoproteasome in line 155? There is no explanation about their role and why the authors refer to them.

-              Line 227: standard of care is the established care, no need for the repetition

-              Single subtitles is the paragraphs do not help. Replace the title and subtitle with a comprehensive title.

Grammar typos and other comments (a few examples):

-              with in line 56

-              line 94

-              in vitro and in vivo should by italics

-              There are a lot of cases that abbreviations are given again and again or are given at a later point

-              while LGG abbreviations are used a lot, high-grade gliomas are not used as abbreviations

Major comments

-It is really difficult to read paragraph 2.1.

-What is the connection of Fig2 with the text?

-In lines 219-226 there is a description of iron's importance but actually, the provided info is really poor. There is a lot of data about ferroptosis and the involvement of iron in GBM. Moreover, ref 33 that is being used in that case is data from neuroblastoma, which is not a brain tumor.

-Within the text, there is spread info about the recurrent state of GBM, without any consistency, while they could be nicely merged.

- Paragraph 5.1 doesn’t explain cell crosstalk. There is just some info about cancer stem cells and some other info about immune cells.

- Paragraph 5.2 just gives one example of the communication of EVs, one example of the role of junctions, and one example of the role of soluble factors. Without any relevant intro or conclusions. It feels that most examples are given without any conclusion or link between them.

- Paragraph 6.2 doesn’t match with its given title

- Paragraph 10 is very confusing and it includes different topics. Also, includes info that could be written earlier in the text.

- Plenty of info in paragraph 10 could be included in the immune paragraph 4 (which is the only paragraph that is well constructed and easy to follow)

- Paragraph 11 could be earlier in the text. There is spread info about different treatment options in the text. Moreover, the paragraph comments also on other treatments and not only on surgery.

- Paragraph 12 as the conclusion refers to the future of glioma research. Nevertheless, there is an extra last paragraph mentions the future of glioma research with a single subtitle. Paragraphs 12 and 13 could be merged.

- Last, but not least: the title. Is the manuscript a deep dive or an overwhelming statement of facts about brain tumors?

Overall, the manuscript needs major re-organization and critical thinking in merging the relevant information. Better introductory and conclusive sentences should be included. Tables could be helpful for data ex genetics ?

Some complexity could be avoided ex legend of figure 2 'the illustration presents a schematic of the' could be Schematic illustration of ...

Some minor grammar typos. A better choice of verbs and syntax could help the reader. 

Author Response

We express our gratitude to Reviewer 3 for the thorough review and valuable criticisms of our manuscript.

1. Organizational and Clarity Enhancements: We have reorganized the manuscript to ensure a logical flow of information and eliminate redundancy. Specific lines and verbs highlighted by you have been revised for better clarity and readability. We have also removed Figure 2 (because it truly didn’t provide the manuscript with any new clarity ) and added a new one in return

2. Accuracy and Repetition: We have corrected inaccuracies and removed repetitive content, like abreviations (for which we have provided a list), ensuring that each section is aligned with the manuscript's overall objectives.

3. We have clarified the roles and significance of the 2021 WHO classification and the immunoproteasome, removing ambiguous references and ensuring all discussed topics are directly relevant to glioblastoma. We have also added more information about ferroptosis.

4. We have removed the subtitles from the manuscript in order to provide a better flow of the text. We have also updated the title. Our manuscript is designed to discuss the key biology behind glioblastoma while taking into account recent discoveries. Moreover, we believe by briefly talking about the other diffuse gliomas, the reader can better understand differences between these diseases.

5. Thanks to your expertise, we have reorganised the manuscript and changed the title for the better. We have adhered to every suggestions you have done and we would like to thank you for taking the time to offer us such an insightful review of our article

Round 2

Reviewer 1 Report

The authors have addressed all issues raised. I have no further requests. 

Author Response

Dear Reviewer,

We appreciate your positive feedback and significant contribution in our manuscript improvement

With high regards,

The collective of authors

Reviewer 2 Report

Authors have replied to all the reviewers comments

Author Response

Dear Reviewer,

We appreciate your positive feedback and significant contribution in our manuscript improvement

With high regards,

The collective of authors

Reviewer 3 Report

Dear Authors,

This version of the manuscript seems structured with clear titles in the text and a clear main title in the manuscript. Table provides a gathered view of the genetic alterations.

Could you please double check that the title and description of the table is in the text.

Figure 3 summarize nicely the intercross of immune cells and GBM cells.

Author Response

Dear Reviewer,

Table title has been added. In the current form of our manuscript, table is now mentioned in the text 

Thank you for your positive feedback

With high regards,

The collective of authors