Extracellular matrices regulate extravasation journey of leukocytes and inflammatory tissue fate

Reviewer #1 (Public review):

Summary:

In this review, the author covered several aspects of the inflammation response, mainly focusing on the mechanisms controlling leukocyte extravasation and inflammation resolution.

Strengths:

This review is based on an impressive number of sources, trying to comprehensively present a very broad and complex topic.

Weaknesses:

(1) This reviewer feels that, despite the title, this review is quite broad and not centred on the role of the extracellular matrix.

(2) The review will benefit from a stronger focus on the specific roles of matrix components and dynamics, with more informative subheadings.

(3) The macrophage phenotype section doesn't seem well integrated with the rest of the review (and is not linked to the ECM).

(4) Table 1 is difficult to follow. It could be reformatted to facilitate reading and understanding

(5) Figure 2 appears very complex and broad.

(6) Spelling and grammar should be thoroughly checked to improve the readability.

Reviewer #2 (Public review):

Summary:

The manuscript is a timely and comprehensive review of how the extracellular matrix (ECM), particularly the vascular basement membrane, regulates leukocyte extravasation, migration, and downstream immune function. It integrates molecular, mechanical, and spatial aspects of ECM biology in the context of inflammation, drawing from recent advances. The framing of ECM as an active instructor of immune cell fate is a conceptual strength.

Strengths:

(1) Comprehensive synthesis of ECM functions across leukocyte extravasation and post-transmigration activity.

(2) Incorporation of recent high-impact findings alongside classical literature.

(3) Conceptually novel framing of ECM as an active regulator of immune function.

(4) Effective integration of molecular, mechanical, and spatial perspectives.

Weaknesses:

(1) Insufficient narrative linkage between the vascular phase (Sections 2-6) and the in-tissue phase (Sections 7-10).

(2) Underrepresentation of lymphocyte biology despite mention in early sections.

(3) The MIKA macrophage identity framework is only loosely tied to ECM mechanisms.

(4) Limited discussion of translational implications and therapeutic strategies.

(5) Overly dense figure insets and underdeveloped links between ECM carryover and downstream immune phenotypes.

(6) Acronyms and some mechanistic details may limit accessibility for a broader readership.

Reviewer #3 (Public review):

Summary & Strengths:

This review by Yu-Tung Li sheds new light on the processes involved in leukocyte extravasation, with a focus on the interaction between leukocytes and the extracellular matrix. In doing so, it presents a fresh perspective on the topic of leukocyte extravasation, which has been extensively covered in numerous excellent reviews. Notably, the role of the extracellular matrix in leukocyte extravasation has received relatively little attention until recently, with a few exceptions, such as a study focusing on the central nervous system (J Inflamm 21, 53 (2024) doi.org/10.1186/s12950-024-00426-6) and another on transmigration hotspots (J Cell Sci (2025) 138 (11): jcs263862 doi.org/10.1242/jcs.263862). This review synthesizes the substantial knowledge accumulated over the past two decades in a novel and compelling manner.

The author dedicates two sections to discussing the relevant barriers, namely, endothelial cell-cell junctions and the basement membrane. The following three paragraphs address how leukocytes interact with and transmigrate through endothelial junctions, the mechanisms supporting extravasation, and how minimal plasma leakage is achieved during this process. The subsequent question of whether the extravasation process affects leukocyte differentiation and properties is original and thought-provoking, having received limited consideration thus far. The consequences of the interaction between leukocytes and the extracellular matrix, particularly regarding efferocytosis, macrophage polarization, and the outcome of inflammation, are explored in the subsequent three chapters. The review concludes by examining tissue-specific states of macrophage identity.

Weaknesses:

Firstly, the first ten sections provide a comprehensive overview of the topic, presenting logical and well-formulated arguments that are easily accessible to a general audience. In stark contrast, the final section (Chapter 11) fails to connect coherently with the preceding review and is nearly incomprehensible without prior knowledge of the author's recent publication in Cell. Mol. Life Sci. CMLS 772 82, 14 (2024). This chapter requires significantly more background information for the general reader, including an introduction to the Macrophage Identity Kinetics Archive (MIKA), which is not even introduced in this review, its basis (meta-analysis of published scRNA-seq data), its significance (identification of major populations), and the reasons behind the revision of the proposed macrophage states and their further development. Secondly, while the attempt to integrate a vast amount of information into fewer figures is commendable, it results in figures that resemble a complex puzzle. The author may consider increasing the number of figures and providing additional, larger "zoom-in" panels, particularly for the topics of clot formation at transmigration hotspots and the interaction between ECM/ECM fragments and integrins. Specifically, the color coding (purple for leukocyte α6-integrins, blue for interacting laminins, also blue for EC α6 integrins, and red for interacting 5-1-1 laminins) is confusing, and the structures are small and difficult to recognize.

Author response:

Public Reviews:

Reviewer #1 (Public review):

Summary:

In this review, the author covered several aspects of the inflammation response, mainly focusing on the mechanisms controlling leukocyte extravasation and inflammation resolution.

Strengths:

This review is based on an impressive number of sources, trying to comprehensively present a very broad and complex topic.

Weaknesses:

(1) This reviewer feels that, despite the title, this review is quite broad and not centred on the role of the extracellular matrix.

(2) The review will benefit from a stronger focus on the specific roles of matrix components and dynamics, with more informative subheadings.

(3) The macrophage phenotype section doesn't seem well integrated with the rest of the review (and is not linked to the ECM).

(4) Table 1 is difficult to follow. It could be reformatted to facilitate reading and understanding

(5) Figure 2 appears very complex and broad.

(6) Spelling and grammar should be thoroughly checked to improve the readability.

This review focuses on the whole extravasation journey of leukocyte and highlights involvement of extracellular matrix (ECM) in multiple phases of the process. ECM may exert their roles either as a collective structure or as individual components. In the revision, for those functions involving specific matrix components, we will emphasize the matrix components and incorporate this information to subheadings as suggested. The parts of macrophage phenotype (Section 10-11) are included for its pivotal roles on deciding the tissue fate following inflammation (ie. to resolve / to regenerate damages incurred or to sustain inflammation), which is an important aspect of this review. ECM could modify macrophage phenotypes either directly (section 10) or indirectly via modulations of tissue stiffness or other cell types like fibroblasts (section 9). However, as pointed out by other reviewers as well, we acknowledge that Section 11 does not integrate well enough to the rest of the review. We plan to reorganize this part and to emphasize its link to ECM during the revision for better integration. We will reformat Table 1 for easier comprehension. We will consider restructuring Figure 2, which outlines various events influencing tissue decision of resolution/inflammation, perhaps by breaking up into two separate figures, to better focus the message. We will also check the language to improve readability.

Reviewer #2 (Public review):

Summary:

The manuscript is a timely and comprehensive review of how the extracellular matrix (ECM), particularly the vascular basement membrane, regulates leukocyte extravasation, migration, and downstream immune function. It integrates molecular, mechanical, and spatial aspects of ECM biology in the context of inflammation, drawing from recent advances. The framing of ECM as an active instructor of immune cell fate is a conceptual strength.

Strengths:

(1) Comprehensive synthesis of ECM functions across leukocyte extravasation and post-transmigration activity.

(2) Incorporation of recent high-impact findings alongside classical literature.

(3) Conceptually novel framing of ECM as an active regulator of immune function.

(4) Effective integration of molecular, mechanical, and spatial perspectives.

Weaknesses:

(1) Insufficient narrative linkage between the vascular phase (Sections 2-6) and the in-tissue phase (Sections 7-10).

(2) Underrepresentation of lymphocyte biology despite mention in early sections.

(3) The MIKA macrophage identity framework is only loosely tied to ECM mechanisms.

(4) Limited discussion of translational implications and therapeutic strategies.

(5) Overly dense figure insets and underdeveloped links between ECM carryover and downstream immune phenotypes.

(6) Acronyms and some mechanistic details may limit accessibility for a broader readership.

We will add a transition paragraph between Section 6 and Section 7 to provide a narrative that the extravasation processes affect downstream leukocyte functions. While lymphocytes follow a similar extravasation principle, their in-tissue activities differ from innate leukocytes. We will thus include discussion of lymphocyte-ECM crosstalk to Section 8 and/or 9 in the revision. We will restructure Section 11 and Figure 3 to better integrate to the rest of the review: In the current manuscript, we merely describe the capability of the MIKA framework to describe identity of any tissue macrophages and thus the framework could serve as a roadmap to facilitate identity normalization of pathological macrophages. We plan, in the revision, by employing the MIKA framework, to discuss and demonstrate linkage between macrophage identities and expression/production of modulators to functional ECM effectors described in Section 8-9. Regarding the comment of limited discussion of translational implications / therapeutic strategies, we will try to enrich this aspect throughout the manuscript where appropriate, in addition to the existing ones (eg. line 293-297; 388-391; 460-463; 512-517) We will also revise figure structure in general to avoid too dense information and to improve clarity. We will consider to provide a glossary explaining specialized terms to expand readership accessibility.

Reviewer #3 (Public review):

Summary & Strengths:

This review by Yu-Tung Li sheds new light on the processes involved in leukocyte extravasation, with a focus on the interaction between leukocytes and the extracellular matrix. In doing so, it presents a fresh perspective on the topic of leukocyte extravasation, which has been extensively covered in numerous excellent reviews. Notably, the role of the extracellular matrix in leukocyte extravasation has received relatively little attention until recently, with a few exceptions, such as a study focusing on the central nervous system (J Inflamm 21, 53 (2024) doi.org/10.1186/s12950-024-00426-6) and another on transmigration hotspots (J Cell Sci (2025) 138 (11): jcs263862 doi.org/10.1242/jcs.263862). This review synthesizes the substantial knowledge accumulated over the past two decades in a novel and compelling manner.

The author dedicates two sections to discussing the relevant barriers, namely, endothelial cell-cell junctions and the basement membrane. The following three paragraphs address how leukocytes interact with and transmigrate through endothelial junctions, the mechanisms supporting extravasation, and how minimal plasma leakage is achieved during this process. The subsequent question of whether the extravasation process affects leukocyte differentiation and properties is original and thought-provoking, having received limited consideration thus far. The consequences of the interaction between leukocytes and the extracellular matrix, particularly regarding efferocytosis, macrophage polarization, and the outcome of inflammation, are explored in the subsequent three chapters. The review concludes by examining tissue-specific states of macrophage identity.

Weaknesses:

Firstly, the first ten sections provide a comprehensive overview of the topic, presenting logical and well-formulated arguments that are easily accessible to a general audience. In stark contrast, the final section (Chapter 11) fails to connect coherently with the preceding review and is nearly incomprehensible without prior knowledge of the author's recent publication in Cell. Mol. Life Sci. CMLS 772 82, 14 (2024). This chapter requires significantly more background information for the general reader, including an introduction to the Macrophage Identity Kinetics Archive (MIKA), which is not even introduced in this review, its basis (meta-analysis of published scRNA-seq data), its significance (identification of major populations), and the reasons behind the revision of the proposed macrophage states and their further development. Secondly, while the attempt to integrate a vast amount of information into fewer figures is commendable, it results in figures that resemble a complex puzzle. The author may consider increasing the number of figures and providing additional, larger "zoom-in" panels, particularly for the topics of clot formation at transmigration hotspots and the interaction between ECM/ECM fragments and integrins. Specifically, the color coding (purple for leukocyte α6-integrins, blue for interacting laminins, also blue for EC α6 integrins, and red for interacting 5-1-1 laminins) is confusing, and the structures are small and difficult to recognize.

We agree with and appreciate the specific and helpful suggestions by the reviewer. During the revision, we will provide the requested background description of MIKA to enhance accessibility of general readership. As pointed out by other reviewers, since this part (Section 11) is less well-integrated to the rest of the review, we will restructure this part by linking tissue macrophage identities under MIKA framework to modulation of functional ECM effectors described in previous sections (Section 8-9). We acknowledge the current figure organization might be overly information-dense and will consider breaking down the contents to multiple figures. The size and color-coding issues will also be addressed.