Understanding the ways that mechanical stimuli influence tumour progression is vital for discovering treatments to overcome the limitations of current options. In this paper, the mathematical models used to quantify and explore the mechanical forces applied to cells and tissues are reviewed. Experiments that attempt to quantify the effects of mechanical stress on different stages of cancer are explored. Lastly, the biological effect of compression on cancer is discussed.

Cancer immunotherapy, specifically Chimeric Antigen Receptor (CAR)-T cell therapy, represents a significant breakthrough in treating cancers. Despite its success in hematological cancers, CAR-T exhibits limited efficacy in solid tumors, which account for more than 90% of all cancers. Solid tumors commonly present unique challenges, including antigen heterogeneity and complex tumor microenvironment (TME). To address these, efforts are being made through improvements in CAR design and the development of advanced validation platforms. While efficacy is limited, some solid tumor types, such as neuroblastoma and gastrointestinal cancers, have shown responsiveness to CAR-T therapy in recent clinical trials. In this review, it is first examined both experimental and computational strategies, such as protein engineering coupled with machine learning, developed to enhance T cell specificity. The challenges and methods associated with T cell delivery and in vivo reprogramming in solid tumors is discussed. It is also explored the advancements in engineered organoid systems, which are emerging as high-fidelity in vitro models that closely mimic the complex human TME and serve as a validation platform for CAR discovery. Collectively, these innovative engineering strategies offer the potential to revolutionize the next generation of CAR-T therapy, ultimately paving the way for more effective treatments in solid tumors.

Skeletal muscle is essential for maintaining body shape and supporting physiological processes. Musculoskeletal function is influenced by various factors, including nutrition, infection, injury or trauma, and advanced glycation end-products (AGEs), which are known to contribute in tissue degeneration, particularly in aging and diabetic populations. This study utilized a skeletal muscle-on-a-chip system to develop three-dimensional in vitro musculoskeletal tissue, enabling a detailed investigation of the effects of AGEs on muscle function and structure. AGEs-induced alterations on muscle were verified by assessments of musculoskeletal contractility, myotube growth, and apoptosis markers. Furthermore, metabolic changes such as modifications in collagen and NADH lifetime changes were observed using fluorescence-lifetime imaging microscopy (FLIM) in the AGEs-treated group. Our result demonstrated a significant reduction in musculoskeletal contractility and structural disruptions in response to AGEs exposure. Overall, these findings provide a robust in vitro model for elucidating the mechanisms by which AGEs impair muscle functionality and integrity, with potential implications for therapeutic strategies aimed at preserving muscle health and enhancing the quality of life in affected populations.

BACKGROUND:Understanding the proteomic-level heterogeneity of the tumor microenvironment (TME) in colorectal cancer (CRC) is crucial due to its well-known heterogeneity. While heterogenous CRC has been extensively characterized at the molecular subtype level, research into the functional heterogeneity of fibroblasts, particularly their relationship with extracellular matrix (ECM) alterations, remains limited. Addressing this gap is essential for a comprehensive understanding of CRC progression and the development of targeted therapies. METHODS:24 tissue samples from 21 CRC patients, along with adjacent normal tissues (NAT), were collected and decellularized using a detergent-based method to enrich the ECM component. Proteomic analysis of ECM-enriched samples was performed using tandem mass tag (TMT) spectrometry, followed by statistical analysis including differential expression protein (DEP) analysis. Single-cell RNA sequencing (scRNA-Seq) data from public datasets were integrated and analyzed to delineate cell states within the TME. Bulk tissue RNA-Seq and bioinformatics analysis, including consensus molecular subtype (CMS) classification and single-cell level deconvolution of TCGA bulk RNA-seq data, were conducted to further explore gene expression patterns and TME composition. RESULTS:Differential cellular origin of the NAT and tumorous ECM proteins were identified, revealing 110 ECM proteins enriched in NAT and 28 ECM proteins in tumor tissues. Desmoplastic and WNT5A+ inflammatory fibroblasts were indicated as the sources of tumor-enriched ECM proteins, while ADAMDEC1+ expressing fibroblasts and PI16+ expressing fibroblast were identified as the sources of NAT-enriched ECM proteins. Deconvolution of bulk RNA-seq of CRC tissues discriminated CMS-specific fibroblast state, reflecting the biological traits of each CMS subtype. Specially, seven ECM genes specific to mesenchymal subtype (CMS4), including PI16+ fibroblast-related 4 genes (SFRP2, PRELP, OGN, SRPX) and desmoplastic fibroblast-related 3 genes (THBS2, CTHRC1, BGN), showed a significant association with poorer survival in patient with CRC. CONCLUSION:We conducted an extracellular matrix (ECM)-focused profiling of the TME by integrating quantitative proteomics with single-cell RNA sequencing (scRNA-seq) data from CRC patients. We identified the ECM proteins of NAT and tumor tissue, and established a cell-matrisome database. We defined mesenchymal subtype-specific molecules associated with specific fibroblast subtypes showing a significant association with poorer survival in patients with CRC. Our ECM-focused profiling of tumor stroma provides new insights as indicators for biological processes and clinical endpoints.

Epithelium layer stands on a membrane, called basement membrane (BM) which serves as a boundary with the underlying stroma. While most studies on morphogenesis have focused on the epithelium-BM complex, the role of the BM-stroma interface remains poorly understood. In this study, we demonstrate how forces originating from the stromal layer contribute to tissue morphogenesis. Folds focalization at the BM-stroma interface is driven by mechanical instability, which arises from the fluidity of the stroma and the polarized tractional forces acting on the rigid membrane of stromal cell condensates. Stromal cells move towards the folded region by topographic guidance, while the concentration of forces intensifies. Through this process, stromal cells and folds engage in recursive interactions, resulting in the formation of a cellular network. Our observation provides a rational mechanism for pattern formation in a multi-layered living tissue. STATEMENT OF SIGNIFICANCE: This study addresses a crucial gap in understanding how stromal cells interact with the basement membrane to lead tissue surface morphogenesis. By developing a collagen-based, nanometer-thick engineered basement membrane, we demonstrate that the stromal cells exert traction forces on the basement membrane to fold. The folding process guides stromal cell migration, which in turn induces further folding in a recursive manner. The direction of folding, invagination or evagination, is determined by the stiffness difference between the stroma and the basement membrane. This model offers better understanding about how the basement membrane interacts with stromal cells to make evaginated network structures on tissue surface.

Cancer metastasis, primarily driven by epithelial-to-mesenchymal transition (EMT), is responsible for most cancer-related mortalities. Traditional pre-clinical models fail to fully capture mesenchymal characteristics due to the loss of human stroma. The extracellular matrix (ECM) plays a crucial role in EMT, yet conventional in vitro models often rely on defined ECM components, which may not adequately replicate the human physiological ECM niche. To mimic the in situ dissemination of cancer cells, we employed a patient-derived extracellular matrix (pdECM). We transitioned the culture matrix for patient-derived colorectal cancer organoids from a basement membrane extract (BME) to a patient-derived ECM (pdECM). We performed single-cell multiomic analyses, integrating transcriptomic and epigenomic data, to investigate changes in organoid phenotypes and reconstruct the EMT trajectory. Organoids cultured in the pdECM exhibited increased tumor cell dissemination and motility, resembling in situ lesions without exogenous ligand treatment. Single-cell multiomic analysis revealed TNF-α signaling as an early metastatic event in the EMT trajectory. Epigenomic changes led to increased accessibility of AP-1 complex target genes, particularly MMP7, which promoted an invasive phenotype. Our multimodal computational approach distinguished early and late EMT states, demonstrating that pdECM-induced EMT occurs independently of traditional EMT master regulators. Notably, pdECM organoids exhibited a partial EMT phenotype, characterized by hybrid epithelial-mesenchymal states. This study presents an advanced in vitro model that closely recapitulates in situ tumorigenesis and provides novel insights into the metastatic cascade. The pdECM system enables the reconstruction of EMT dynamics, highlighting the critical role of ECM composition in metastasis and offering a physiologically relevant platform for the development of targeted therapies.

Gastric cancer (GC) is a highly heterogeneous disease regarding histologic features, genotypes, and molecular phenotypes. Here, we investigate extracellular matrix (ECM)-centric analysis, examining its association with histologic subtypes and patient prognosis in human GC. We performed quantitative proteomic analysis of decellularized GC tissues that characterizes tumorous ECM, highlighting proteomic heterogeneity in ECM components. We identified 20 tumor-enriched proteins including four glycoproteins, serpin family H member 1 (SERPINH1), annexin family (ANXA3/4/5/13), S100A family (S100A6/8/9), MMP14, and other matrisome-associated proteins. In addition, histopathological characteristics of GC reveals differential expression in ECM composition, with the poorly cohesive carcinoma-not otherwise specified (PCC-NOS) subtype being distinctly demarcated from other histologic subtypes. Integrating ECM proteomics with single-cell RNA sequencing, we identified crucial molecular markers in the PCC-NOS-specific stroma. PCC-NOS-enriched matrisome proteins and gene expression signatures of adipogenic cancer-associated fibroblasts (CAFadi) are closely linked, both associated with adverse outcomes in GC. Using tumor microarray analysis, we confirmed the CAFadi surface marker, ATP binding cassette subfamily A member 8 (ABCA8), predominantly present in PCC-NOS tumors. Our ECM-focused analysis paves the way for studies to determine their utility as biomarkers for patient stratification, offering valuable insights for linking molecular and histologic features in GC.

Conventional tumor models have critical shortcomings in that they lack the complexity of the human stroma. The heterogeneous stroma is a central compartment of the tumor microenvironment (TME) that must be addressed in cancer research and precision medicine. To fully model the human tumor stroma, the deconstruction and reconstruction of tumor tissues have been suggested as new approaches for in vitro tumor modeling. In this review, we summarize the heterogeneity of tumor-associated stromal cells and general deconstruction approaches used to isolate patient-specific stromal cells from tumor tissue; we also address the effect of the deconstruction procedure on the characteristics of primary cells. Finally, perspectives on the future of reconstructed tumor models are discussed, with an emphasis on the essential prerequisites for developing authentic humanized tumor models.

Advanced glycation end products (AGEs) play a pivotal role in the aging process, regarded as a hallmark of aging. Despite their significance, the absence of adequate monitoring tools has hindered the exploration of the relationship between AGEs and aging. Here, we present a novel AGE-selective probe, AGO, for the first time. AGO exhibited superior sensitivity in detecting AGEs compared to the conventional method of measuring autofluorescence from AGEs. Furthermore, we validated AGO's ability to detect AGEs based on kinetics, demonstrating a preference for ribose-derived AGEs. Lastly, AGO effectively visualized glycation products in a collagen-based mimicking model of glycation. We anticipate that this study will enhance the molecular tool sets available for comprehending the physiological processes of AGEs during aging.

No standardized in vitro cell culture models for glioblastoma (GBM) have yet been established, excluding the traditional two-dimensional culture. GBM tumorspheres (TSs) have been highlighted as a good model platform for testing drug effects and characterizing specific features of GBM, but a detailed evaluation of their suitability and comparative performance is lacking. Here, we isolated GBM TSs and extracellular matrices (ECM) from tissues obtained from newly diagnosed IDH1 wild-type GBM patients and cultured GBM TSs on five different culture platforms: (1) ordinary TS culture liquid media (LM), (2) collagen-based three-dimensional (3D) matrix, (3) patient typical ECM-based 3D matrix, (4) patient tumor ECM-based 3D matrix, and (5) mouse brain. For evaluation, we obtained transcriptome data from all cultured GBM TSs using microarrays. The LM platform exhibited the most similar transcriptional program to paired tissues based on GBM genes, stemness- and invasiveness-related genes, transcription factor activity, and canonical signaling pathways. GBM TSs can be cultured via an easy-to-handle and cost- and time-efficient LM platform while preserving the transcriptional program of the originating tissues without supplementing the ECM or embedding it into the mouse brain. In addition to applications in basic cancer research, GBM TSs cultured in LM may also serve as patient avatars in drug screening and pre-clinical evaluation of targeted therapy and as standardized and clinically relevant models for precision medicine.

The solid tumor microenvironment presents many challenges for immunotherapy that must be addressed for clinical relevance. This review will highlight research that has attempted to tackle these challenges and summarize recent therapeutic approaches to enhance both native and engineered T cell functions through hydrogel delivery. Different types of hydrogels are presented in this paper and organized based on purpose of the research: activation and expansion, delivery methods, co-delivery of immunostimulating agents, and overcoming tumor microenvironment barriers. Perspectives in the future of solid cancer immunotherapy are also discussed to share opinion on necessary research for the success in this field.

S1. Elevated c-MYC activity in MCF7-BrM promotes brain metastasis (related to Fig 1) S2. C-MYC regulates tumor sphere circularity and invasiveness in a HA-collagen matrix (related to Fig 2) S3. c-MYC-mediated recruitment of RAW cells (related to Fig 3) S4. qRT-PCR analysis of c-MYC-regulated genes (related to Fig 4) S5. Association of c-MYC and MYC-BrMFS with BrMFS in the MSK82 dataset (related to Fig 5) S6. BrM-BCCs exhibit elevated TRAIL sensitivity (related to Fig 6)

Collagen is a prominent target of nonenzymatic glycation, which is a hallmark of aging and causes functional alteration of the matrix. Here, we uncover glycation-mediated structural and functional changes in the collagen-enriched meningeal membrane of the human and mouse brain. Using an in vitro culture platform mimicking the meningeal membrane composed of fibrillar collagen, we showed that the accumulation of advanced glycation end products (AGEs) in the collagen membrane is responsible for glycation-mediated matrix remodeling. These changes influence fibroblast-matrix interactions, inducing cell-mediated ECM remodeling. The adherence of meningeal fibroblasts to the glycated collagen membrane was mediated by the discoidin domain-containing receptor 2 (DDR2), whereas integrin-mediated adhesion was inhibited. A-kinase anchoring protein 12 (AKAP12)-positive meningeal fibroblasts in the meningeal membrane of aged mice exhibited substantially increased expression of DDR2 and depletion of integrin beta-1 (ITGB1). In the glycated collagen membrane, meningeal fibroblasts increased the expression of matrix metalloproteinase 14 (MMP14) and less tissue inhibitor of metalloproteinase-1 (TIMP1). In contrast, the cells exhibited decreased expression of type I collagen (COL1A1). These results suggest that glycation modification by meningeal fibroblasts is intimately linked to aging-related structural and functional alterations in the meningeal membrane.

Cancer secretome is a reservoir for aberrant glycosylation. How therapies alter this post- translational cancer hallmark and the consequences thereof remain elusive. Here, we show that an elevated secretome fucosylation is a pan-cancer signature of both response and resistance to multiple targeted therapies. Large-scale pharmacogenomics revealed that fucosylation genes display widespread association with resistance to these therapies. In cancer cell cultures, xenograft mouse models, and patients, targeted kinase inhibitors distinctively induced core fucosylation of secreted proteins less than 60 kDa. Label-free proteomics of N-glycoproteomes identified fucosylation of the antioxidant PON1 as a critical component of the therapy-induced secretome (TIS). N-glycosylation of TIS and target core fucosylation of PON1 are mediated by the fucose salvage-FUT8-SLC35C1 axis with PON3 directly modulating GDP-Fuc transfer on PON1 scaffolds. Core fucosylation in the Golgi impacts PON1 stability and folding prior to secretion, promoting a more degradation-resistant PON1. Global and PON1-specific secretome de-N-glycosylation both limited the expansion of resistant clones in a tumor regression model. We defined the resistance-associated transcription factors (TFs) and genes modulated by the N-glycosylated TIS via a focused and transcriptome-wide analyses. These genes characterize the oxidative stress, inflammatory niche, and unfolded protein response as important factors for this modulation. Our findings demonstrate that core fucosylation is a common modification indirectly induced by targeted therapies that paradoxically promotes resistance.

Cell-generated mechanical forces drive many cellular and tissue-level movements and rearrangements required for the tissue or organ to develop its shape1, 2, 3, 4, 5. The prevalent view of tissue morphogenesis relies on epithelial folding resulting in compressed epithelial monolayers, overlooking the involvement of stroma in morphogenesis1, 4, 6, 7. Here, we report a giant web-like network formation of stromal cells in the epithelium-stroma interface, resulting from a multi-scale mechano-reciprocity between migrating cells and their extracellular environment. In multi-layered tissues, surface wrinkles form by a stromal cell-mediated tensional force exerted at the basement membrane. The topographical cue is transmitted to the stromal cell, directing its protrusion and migration along the wrinkles. This inductive movement of the cells conveys traction forces to its surrounding extracellular matrix, remodeling the local architectures of the stroma. In this manner, stromal cells and wrinkles communicate recursively to generate the cellular network. Our observation provides a rational mechanism for network formation in living tissues and a new understanding of the role of cellular-level tensional force in morphogenesis.

Biological systems are composed of hierarchical structures made of a large number of proteins. These structures are highly sophisticated and challenging to replicate using artificial synthesis methods. To exploit these structures in materials science, biotemplating is used to achieve biocomposites that accurately mimic biological structures and impart functionality of inorganic materials, including electrical conductivity. However, the biological scaffolds used in previous studies are limited to stereotypical and simple morphologies with little synthetic diversity because of a lack of control over their morphologies. This study proposes that the specific protein assemblies within the cell-derived extracellular matrix (ECM), whose morphological features are widely tailorable, can be employed as versatile biotemplates. In a typical procedure, a fibrillar assembly of fibronectin-a constituent protein of the ECM-is metalized through an antibody-guided biotemplating approach. Specifically, the antibody-bearing nanogold is attached to the fibronectin through antibody-antigen interactions, and then metals are grown on the nanogold acting as a seed. The biomimetic structure can be adapted for hydrogen production and sensing after improving its electrical conductivity through thermal sintering or additional metal growth. This study demonstrates that cell-derived ECM can be an attractive option for addressing the diversity limitation of a conventional biotemplate.

The extracellular matrix (ECM) of the tumor microenvironment undergoes constant remodeling that alters its biochemical and mechano-physical properties. Non-enzymatic glycation can induce the formation of advanced glycation end-products (AGEs), which may cause abnormal ECM turnover with excessively cross-linked collagen fibers. However, the subsequent effects of AGE-mediated matrix remodeling on the characteristics of stromal cells in tumor microenvironments remain unclear. Here, we demonstrate that AGEs accumulated in the ECM alter the fibroblast phenotype within a three-dimensional collagen matrix. Both the AGE interaction with its receptor (RAGE) and integrin-mediated mechanotransduction signaling were up-regulated in glycated collagen matrix, leading to fibroblast activation to acquire a cancer-associated fibroblast (CAF)-like phenotype. These effects were blocked with neutralizing antibodies against RAGE or the inhibition of focal adhesion (FA) signaling. An AGE cross-link breaker, phenyl-4,5dimethylthiazolium bromide (ALT 711), also reduced the transformation of fibroblasts into the CAF-like phenotype because of its dual inhibitory role in the AGE-modified matrix. Apart from targeting the AGE-RAGE interaction directly, the decreased matrix stiffness attenuated fibroblast activation by inhibiting the downstream cellular response to matrix stiffness. Our results suggest that indirect/direct targeting of accumulated AGEs in the ECM has potential for targeting the tumor stroma to improve cancer therapy. Statement of significance Advanced glycated end-products (AGEs)-modified extracellular matrix (ECM) is closely associated with pathological states and is recognized as a critical factor that precedes tumorigenesis. While increased matrix stiffness is known to induce fibroblast activation, less is known about how both biochemical and mechano-physical changes in AGE-mediated matrix-remodeling cooperate to produce a myofibroblastic cancer-associated fibroblast (CAF)-like phenotype. For the first time, we found that both the AGE interaction with its receptor (RAGE) and integrin-mediated mechanotransduction were up-regulated in glycated collagen matrix, leading to fibroblast activation. We further demonstrated that an AGE cross-link breaker, ALT-711, reduced the CAF-like transformation because of its dual inhibitory role in the AGE-modified matrix. Our findings offer promising extracellular-reversion strategies targeting the non-enzymatic ECM glycation, to regulate fibroblast activation. (C) 2022 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

When embedded into a three-dimensional (3D) matrix, cancer stem cells (or cancer-initiating cells) can grow into self-organizing organotypic structures called tumor organoids. During organoid formation, the matrix not only provides structural support but also delivers biochemical signals. Although increasing evidence indicates that the extracellular matrix (ECM) is an essential component of the tumor microenvironment during tumor development and progression, the influence of the ECM on organoid formation has been largely ignored; the ECM has only recently been recognized to play a role in the regulation of cancer cell phenotypes. We reviewed ECM-based hydrogels to tailoring tumor organoids and highlight the potential role of the ECM in the development of recapitulating malignant/invasive tumor organoids with enhanced capacity for in vitro representation of ECM-regulated tumor progression.