Review Reports - Glioblastoma Stem Cells as Targets for Emerging Precision Immunotherapies and Molecular Treatments

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

To the Authors

Thank you for submitting this commentary entitled “Glioblastoma and Other Cancer Stem Cells: Targets for Emerging Precision Immuno- and Molecular Therapies.” This article presents a broad overview of the development of the neural stem cell concept, its relationship to glioma and glioblastoma stem-like cells, and the therapeutic implications of these ideas in the era of precision oncology. The manuscript also highlights recent advances in single-cell and spatial technologies, immunotherapy, molecular targeting, and artificial intelligence as promising tools for improving our understanding and treatment of glioblastoma. The overall theme is timely and relevant, and the article reflects the author’s long-standing engagement with this field.

Overall, I found this to be a thoughtful and interesting commentary, and I would support acceptance after minor revision. The manuscript succeeds in conveying a broad conceptual perspective and brings together historical and emerging aspects of cancer stem cell biology in an engaging manner. It is likely to be of interest to readers in neuro-oncology, especially those interested in translational and future-oriented research directions.

I would suggest three minor points that may help strengthen the manuscript further. First, it would be helpful to clarify the scope of the article more explicitly. Although the title refers to “glioblastoma and other cancer stem cells,” the discussion is predominantly focused on glioma and glioblastoma. A brief clarification of whether the broader cancer stem cell framework is intended mainly as context, rather than as an equal focus of the article, would improve alignment between the title and the text.

Second, the article would benefit from a slightly more balanced discussion of remaining uncertainties. The commentary appropriately mentions ongoing controversy regarding the precise cells of origin of certain glioma subtypes and acknowledges that major translational challenges remain. Expanding this balanced perspective just a little would make the manuscript even stronger, particularly in sections that discuss therapeutic promise.

Third, the concluding discussion of AI, machine learning, and future therapeutic strategies is stimulating, but it could be tightened to improve focus. A more direct connection between the biological concepts reviewed earlier and the practical clinical or translational implications in the final section would help the paper finish on an even stronger note.

In summary, this is a well-conceived commentary on a timely topic. With minor revision to improve scope clarity, balance, and focus, I believe the manuscript would be suitable for publication.

Author Response

Reviewer 1 comment:
I would suggest three minor points that may help strengthen the manuscript further. First, it would be helpful to clarify the scope of the article more explicitly. Although the title refers to “glioblastoma and other cancer stem cells,” the discussion is predominantly focused on glioma and glioblastoma. A brief clarification of whether the broader cancer stem cell framework is intended mainly as context, rather than as an equal focus of the article, would improve alignment between the title and the text.

Response:

We thank the reviewer for these helpful and constructive suggestions, which we believe have improved the clarity and balance of the commentary.

1. Clarification of scope and title alignment:
We agree that the manuscript is predominantly focused on glioblastoma and glioblastoma stem‑like cellular states, with references to other cancer stem cell systems included primarily for historical and conceptual context. In response to this comment and similar feedback from other reviewers, we have revised the title to explicitly emphasize glioblastoma stem cells, thereby removing ambiguity regarding the scope of the article. In addition, we have ensured that mentions of broader cancer stem cell frameworks are clearly framed as contextual background rather than as an equal focus of the commentary.

Reviewer 1 comment:
Second, the article would benefit from a slightly more balanced discussion of remaining uncertainties. The commentary appropriately mentions ongoing controversy regarding the precise cells of origin of certain glioma subtypes and acknowledges that major translational challenges remain. Expanding this balanced perspective just a little would make the manuscript even stronger, particularly in sections that discuss therapeutic promise.

2. Balanced discussion of remaining uncertainties and translational challenges:
We agree that a balanced perspective is important, particularly when discussing emerging therapeutic strategies. In the revised manuscript, we have further emphasized ongoing uncertainties, including the debated cells of origin of specific glioma subtypes, the dynamic and plastic nature of stem‑like cellular states and persistent barriers to successful clinical translation. These considerations are now more clearly integrated into sections discussing therapeutic promise, reinforcing that precision molecular and immunotherapeutic advances must contend with tumor heterogeneity, microenvironmental influences, and evolutionary adaptation.

We believe these revisions strengthen the manuscript by clarifying its scope and maintaining an appropriately critical and balanced tone while highlighting future therapeutic opportunities. We thank the reviewer for these thoughtful suggestions.

Reviewer 1 comment:
The concluding discussion of AI, machine learning, and future therapeutic strategies is stimulating, but it could be tightened to improve focus. A more direct connection between the biological concepts reviewed earlier and the practical clinical or translational implications in the final section would help the paper finish on an even stronger note.

Response:

We thank the reviewer for this helpful comment and positive assessment of the concluding section. In response, we have refined the final section of the manuscript to make more explicit the connection between the biological concepts discussed earlier—particularly glioblastoma stem‑like cellular states, lineage plasticity, spatial heterogeneity, and microenvironmental interactions—and the role of AI‑driven approaches in future translational and clinical applications.

Specifically, we have added a clarifying sentence at the beginning of Section 5 emphasizing that the adaptive, spatially structured nature of glioblastoma stem‑like states generates biological complexity that exceeds the capacity of linear biomarker‑based approaches, thereby providing a clear rationale for the use of AI‑driven integrative models. This addition directly links the stem cell–centric and microenvironmental biology reviewed in earlier sections to the need for machine‑learning frameworks capable of integrating multiscale, longitudinal and spatial data in support of precision molecular and immunotherapeutic strategies.

We believe this revision tightens the focus of the concluding discussion and more clearly demonstrates how the biological principles developed throughout the commentary inform practical translational directions.

Reviewer 2 Report

Comments and Suggestions for Authors

This manuscript presents an interesting commentary on glioblastoma stem cells and emerging precision therapies. However, a few issues should be addressed before publication:

The manuscript focuses almost entirely on glioblastoma, with only a brief mention of breast, osteosarcoma, colon, and prostate cancers in section 2. Please revise the title to accurately focus on glioblastoma stem cells.
The title emphasizes cancer stem cells as targets for emerging molecular and immunotherapies; however, the discussion of these approaches in section 3 is too generalized. Please explain exactly how these therapies specifically target CSCs. Moreover, please include a summary diagram illustrating how the discussed molecular and immunotherapies target glioblastoma stem cells.
In line 124, please provide the appropriate reference for the SeekRNA technology.
Please check the reference list carefully for formatting consistency and typos. For example, specific formatting issues were found in ref. 9, 13, 43, and 45.

Author Response

Reviewer 2 comment:
The manuscript focuses almost entirely on glioblastoma, with only a brief mention of breast, osteosarcoma, colon, and prostate cancers in Section 2. Please revise the title to accurately focus on glioblastoma stem cells.

Response:

We thank the reviewer for this helpful suggestion. We agree that the primary focus of the commentary is glioblastoma and that other tumor types were mentioned only briefly to provide historical context for the cancer stem cell concept.

In direct response to this comment, we have revised the title from:

“Glioblastoma and Other Cancer Stem Cells: Targets for Emerging Precision Immuno‑ and Molecular Therapies”

to:

“Glioblastoma Stem Cells as Targets for Emerging Precision Immuno‑therapies and Molecular Treatments.”

This revised title more accurately reflects the scope and emphasis of the manuscript and removes any potential ambiguity regarding coverage of non‑glioblastoma tumor types. We have also carefully reviewed the text to ensure that references to other cancers remain clearly contextual and do not distract from the glioblastoma‑focused narrative.

Reviewer 2 comment:
Please explain how these therapies specifically target CSCs and include a summary diagram.

Response:

To address this request, we have clarified how emerging molecular and immunotherapeutic strategies are specifically informed by glioblastoma stem cell (GSC) biology. In the revised manuscript, GSCs are framed as stem‑like cellular states sustained by intrinsic signaling pathways, immune‑evasive mechanisms, and supportive microenvironmental niches.

To further improve clarity, we have added Figure 2, which schematically illustrates how contemporary precision molecular therapies, immune‑based strategies, and niche‑ and system‑level modulators converge on glioblastoma stem‑like cellular states. Together with the revised text in Section 4, this figure explicitly demonstrates how these therapies are designed to target GSC‑associated vulnerabilities rather than bulk tumor populations.

Reviewer 2 comment:
In line 124, please provide the appropriate reference for the seekRNA technology. Please check the reference list carefully for formatting consistency and typos.

Response:

We have updated the reference list for formatting consistency and typos and have included the reference for the seekRNA technology (ref 44, line 362)

Reviewer 3 Report

Comments and Suggestions for Authors

This report is a short commentary about GBM and other cancer stem cells as a target for precision immuno- and molecular therapies. The themes must be important when we think about the future of GBM-related therapies. I would like to raise the following comments about it.

<Major>

The main theme of this commentary is comprised of GBM (cancer) stem cells and precision immuno- and molecular therapies. In a general sense, the issue of GBM (cancer) stem cells, which covers the former half of the MS, is an old one while that of precision immuno- and molecular therapies, which covers the latter half of it, is a new one. As a result, the former half of the reference is dominated by old references, and the latter half of it, by new references. This may be okay, but we know that the concept of GBM (cancer) stem cells have ever been changing since its introduction. There are many recent papers addressing, for example, de-differentiated cancer stem cells from non-cancer stem cells, cancer stem cells mimicking normal stem cells to try to escape from therapies, or a bunch of active interactions between GBM (cancer) stem cells and tumor microenvironment. Particularly the third one may be important to understand the latter theme of this paper, ‘precision immuno- and molecular therapies.’ But I could not find a concise and yet a sufficient description about the third one. I think a certain volume of gap, content-wise, is present between the Chapter 2 and Chapter 3, because of which it is difficult to collectively understand GBM (cancer) stem cells and precision immuno- and molecular therapies. After all, how are they related? I would like to suggest the author to comment on the latest data or knowledge about GBM (cancer) stem cells, make the references updated, and make the issue of how GBM (cancer) stem cells are related to precision immuno- and molecular therapies more understandable.

<Minor>

line 52, ‘a stage 4 glioma’: This word is understandable, but strictly speaking, this should be ‘a (WHO) grade 4 glioma.’

Author Response

Reviewer 3 comment:

Response:

We thank the reviewer for this insightful and constructive comment. We fully agree that the concept of glioblastoma (GBM) stem cells has evolved substantially since its initial introduction and that contemporary views emphasize cellular plasticity, de‑differentiation, and dynamic interactions with the tumor microenvironment rather than a fixed cancer stem cell hierarchy.

In response, we have revised the manuscript to more explicitly reflect these advances and to clarify how evolving concepts of GBM stem cell biology directly inform precision immuno‑ and molecular therapeutic strategies. Specifically:

Updated conceptual framework of GBM stem cells:
Throughout the revised manuscript, GBM stem cells are now framed as stem‑like cellular states rather than a static population. We emphasize lineage plasticity, clonal evolution and the capacity of non‑stem tumor cells to reacquire stem‑like features under microenvironmental and therapeutic pressures, incorporating recent single‑cell and spatial transcriptomic evidence.
Expanded discussion of tumor microenvironment interactions:
We have strengthened discussion of active, bidirectional interactions between GBM stem‑like states and the tumor microenvironment, including immune, vascular, myeloid, neuronal, and systemic niches. These interactions are now integrated into both the biological narrative (Sections 3–4) and the therapeutic framework, highlighting their role in therapeutic resistance and disease progression.
Clarified linkage between GBM stem cell biology and precision therapies:
To address the perceived content gap between early sections on GBM stem cells and later sections on precision therapies, we introduce a unifying conceptual framework linking ontogeny, molecular biography, and therapeutic vulnerability (Figure 1). In addition, Figure 2 explicitly illustrates how contemporary precision molecular, immune‑based, and niche‑modulating therapies converge on GBM stem‑like cellular states. These additions are intended to make the relationship between GBM stem cell biology and precision immuno‑ and molecular therapies more explicit and accessible.
Updated and expanded references:
We have updated the reference list to include recent studies addressing GBM stem cell plasticity, microenvironmental regulation, single‑cell and spatial profiling, and therapeutic modulation of stem‑like states, ensuring that both foundational and emerging concepts are appropriately represented.

We believe these revisions resolve the content gap noted by the reviewer and clarify how modern interpretations of GBM stem cell biology provide the conceptual basis for emerging precision immuno‑ and molecular therapeutic approaches. We thank the reviewer again for their valuable suggestions, which have strengthened the clarity and impact of the commentary.

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The authors addressed all my comments. However, I still have one minor suggestion: Figure 2 is a little too simple. It would be great to recreate this figure using a tool like BioRender, which will make the manuscript look more engaging and professional.

Author Response

Reviewer 2 minor comment:

We thank the reviewer for this helpful suggestion. Figure 2 has been substantially revised to increase its visual and conceptual richness. The updated schematic now illustrates the convergence of intrinsic targeting, immune‑mediated killing, and niche‑ and system‑level modulation on glioblastoma stem‑like cellular states, incorporating clearer mechanistic categories and refined graphical elements. We believe the revised Figure 2 now more effectively conveys the complexity and integration of contemporary precision strategies targeting glioblastoma stem‑like states. Changes are highlighted in the revised Figure 2 and accompanying legend.