A spheroid whole mount drug testing pipeline with machine-learning based image analysis identifies cell-type specific differences in drug efficacy on a single-cell level DOI Creative Commons
Mario Vitacolonna, Roman Bruch,

Richard Schneider

et al.

BMC Cancer, Journal Year: 2024, Volume and Issue: 24(1)

Published: Dec. 18, 2024

Abstract Background The growth and drug response of tumors are influenced by their stromal composition, both in vivo 3D-cell culture models. Cell-type inherent features as well mutual relationships between the different cell types a tumor might affect susceptibility whole and/or its populations. However, lack single-cell procedures with sufficient detail has hampered automated observation cell-type-specific effects three-dimensional stroma-tumor co-cultures. Methods Here, we developed high-content pipeline ranging from setup novel tumor-fibroblast spheroid co-cultures over optical tissue clearing, mount staining, 3D confocal microscopy to optimized 3D-image segmentation 3D-deep-learning model automate analysis range processes, such proliferation, apoptosis, necrosis, susceptibility, nuclear morphology, density. Results This demonstrated that KP-4 cells CCD-1137Sk fibroblasts exhibited advantage compared mono-cultures, resulting higher counts following cytostatic treatments paclitaxel doxorubicin. revealed this apparent benefit was due resilience against drugs did not indicate resistance cancer during co-culture. Conversely, were partially even more susceptible presence than mono-cultures. Conclusion In summary, underlines method can reveal critical insights regarding mechanism action substances

Language: Английский

The Role of Fibroblasts in Skin Homeostasis and Repair DOI Creative Commons
Federica Boraldi, Francesco Demetrio Lofaro,

Susanna Bonacorsi

et al.

Biomedicines, Journal Year: 2024, Volume and Issue: 12(7), P. 1586 - 1586

Published: July 17, 2024

Fibroblasts are typical mesenchymal cells widely distributed throughout the human body where they (1) synthesise and maintain extracellular matrix, ensuring structural role of soft connective tissues; (2) secrete cytokines growth factors; (3) communicate with each other cell types, acting as signalling source for stem niches; (4) involved in tissue remodelling, wound healing, fibrosis, cancer. This review focuses on developmental heterogeneity dermal fibroblasts, their ability to sense changes biomechanical properties surrounding aging, skin repair, pathologic conditions tumour development. Moreover, we describe use fibroblasts different models (e.g., vivo animal vitro systems from 2D 6D cultures) bioengineering informative potential high-throughput assays study under disease contexts personalized healthcare regenerative medicine applications.

Language: Английский

Citations

16
Post-Translational Modifications of Proteins Orchestrate All Hallmarks of Cancer DOI Creative Commons
Pathea Bruno,

Aneeta Arshad,

Maria-Raluca Gogu

et al.

Life, Journal Year: 2025, Volume and Issue: 15(1), P. 126 - 126

Published: Jan. 18, 2025

Post-translational modifications (PTMs) of proteins dynamically build the buffering and adapting interface between oncogenic mutations environmental stressors, on one hand, cancer cell structure, functioning, behavior. Aberrant PTMs can be considered as enabling characteristics long they orchestrate all malignant variability in proteome cells, cancer-associated tumor microenvironment (TME). On other enhance anticancer mechanisms tumoral ecosystem or sustain beneficial effects oncologic therapies through degradation inactivation carcinogenic or/and activation tumor-suppressor proteins. In this review, we summarized analyzed a wide spectrum involved regulatory that drive tumorigenesis, genetic instability, epigenetic reprogramming, events metastatic cascade, cytoskeleton extracellular matrix (ECM) remodeling, angiogenesis, immune response, tumor-associated microbiome, metabolism rewiring most important hallmarks cancer. All develop due to proteins, which modulate gene transcription, intracellular signaling, protein size, activity, stability localization, trafficking, secretion, half-life, protein–protein interactions (PPIs). associated with exploited better understand underlying molecular heterogeneous chameleonic disease, find new biomarkers progression prognosis, personalize oncotherapies, discover targets for drug development.

Language: Английский

Citations

2
ECM, integrins, and DDRs: A nexus of cancer progression, therapy, and future directions DOI
M A Azim, Julie S. Di Martino

Matrix Biology, Journal Year: 2025, Volume and Issue: 138, P. 27 - 43

Published: April 12, 2025

Language: Английский

Citations

0
Invasive cancer cells soften collagen networks and disrupt stress-stiffening via volume exclusion, contractility and adhesion DOI Creative Commons

Irène Nagle,

Margherita Tavasso, Ankur Bordoloi

et al.

Published: April 17, 2025

Abstract Collagen networks form the structural backbone of extracellular matrix in both healthy and cancerous tissues, exhibiting nonlinear mechanical properties that crucially regulate tissue mechanics cell behavior. Here, we investigate how presence invasive breast cancer cells (MDA-MB-231) influences polymerization kinetics collagen using bulk shear rheology rheo-confocal microscopy. We show embedded delay onset due to volume exclusion effects. During polymerization, (at 4% fraction) cause an unexpected time-dependent softening network. this effect arises from active remodeling via adhesion contractility rather than proteolytic degradation. At higher fractions, dominant shifts exclusion, causing a two-fold reduction network stiffness. Additionally, demonstrate suppress characteristic stress-stiffening response collagen. This (partially) disappears when are inhibited, it is absent replaced by passive hydrogel particles. These findings provide new insights into inclusions modify fibrous networks, contributing better understanding role diseased tissues like tumors. Graphical Highlights Using biomimetic model based on containing cells, these soften impair stress-stiffening. By comparing impact cell-sized beads, cell-matrix adhesion, have distinct contributions changes networks.

Language: Английский

Citations

0
Tumoroids, a valid preclinical screening platform for monitoring cancer angiogenesis DOI Creative Commons
Zahra Abbasi‐Malati,

Parisa Khanicheragh,

Maryam Taghavi Narmi

et al.

Stem Cell Research & Therapy, Journal Year: 2024, Volume and Issue: 15(1)

Published: Aug. 26, 2024

In recent years, biologists and clinicians have witnessed prominent advances in vitro 3D culture techniques related to biomimetic human/animal tissue analogs. Numerous data confirmed that unicellular multicellular (tumoroids) tumor spheroids with dense native cells certain matrices are sensitive valid analytical tools for drug screening, cancer cell dynamic growth, behavior, etc. laboratory settings. Angiogenesis/vascularization is a very critical biological phenomenon support oxygen nutrients within the deep layer of solid masses. It has been shown endothelial (EC)-incorporated or -free spheroid/tumoroid systems provide relatively reliable platform monitoring formation nascent blood vessels micron/micrometer scales. Besides, paracrine angiogenic activity can be monitored after being treated different therapeutic approaches. Here, we aimed collect findings angiogenesis using spheroids. Vascularized spheroids/tumoroids help us elucidation mechanisms formation, development, metastasis by main influencing factors.

Language: Английский

Citations

2
Recombinant humanized type III collagen inhibits ovarian cancer and induces protective anti-tumor immunity by regulating autophagy through GSTP1 DOI Creative Commons
Hui Zeng, Hu Li, Li Wang

et al.

Materials Today Bio, Journal Year: 2024, Volume and Issue: 28, P. 101220 - 101220

Published: Sept. 1, 2024

Ovarian cancer (OC) is one of the leading causes death from malignancy in women and lacks safe efficient treatment. The novel biomaterial, recombinant humanized collagen type III (rhCOLIII), has been reported to have various biological functions, but its role OC unclear. This study aimed reveal function mechanism action rhCOLIII OC. We developed an injectable human (rhCOL)-derived material with a molecular weight 45 kDa, stable triple helix structure, high biocompatibility, water solubility biosafety. anti-tumor activity was comprehensively evaluated through

Language: Английский

Citations

2
A Fibroblast-Derived Secretome Stimulates the Growth and Invasiveness of 3D Plexiform Neurofibroma Spheroids DOI Open Access
Kyungmin Ji,

George J. Schwenkel,

Raymond R. Mattingly

et al.

Cancers, Journal Year: 2024, Volume and Issue: 16(14), P. 2498 - 2498

Published: July 9, 2024

Plexiform neurofibromas (PNs) occur in about a half of neurofibromatosis type 1 (NF1) patients and have garnered significant research attention due to their capacity for growth potential malignant transformation. NF1 plexiform neurofibroma (pNF1) is complex tumor composed Schwann cell-derived cells (Nf1−/−) the microenvironment (TME). Although it has been widely demonstrated that TME involved formation neurofibromas, little known effects on subsequent progression human pNF1. Elucidating molecular interactions between may provide new therapeutic targets reduce In present study, we focused contributions fibroblasts, most abundant cell types TME, To simulate used three-dimensional (3D) coculture model immortalized pNF1 primary fibroblasts (Nf1+/−) derived from patients. We performed live-cell imaging 3D/4D (3D real-time) cultures through confocal microscopy followed by 3D quantitative analyses using advanced software. The spheroids cocultures with was significantly greater than monocultures. An increase also occurred when they were cultured conditioned media (CM) fibroblasts. Moreover, fibroblast-derived CM increased invasive outgrowth further local invasion spheroids. Interestingly, small extracellular vesicles (sEVs) depleted CM, stimulation lost. Our results suggest sEVs are target reducing

Language: Английский

Citations

1
Overexpression of Basonuclin Zinc Finger Protein 2 in stromal cell is related to mesenchymal phenotype and immunosuppression of mucinous colorectal adenocarcinoma DOI Creative Commons

Qing-Zhong Yin,

Yuan-Jie Liu,

Qian Zhang

et al.

International Immunopharmacology, Journal Year: 2024, Volume and Issue: 142, P. 113184 - 113184

Published: Sept. 21, 2024

Language: Английский

Citations

0
Identification of SLC22A17 DNA methylation hotspot as a potential biomarker in cutaneous melanoma DOI Creative Commons
Alessandro Lavoro, Luca Falzone, Giuseppe Gattuso

et al.

Journal of Translational Medicine, Journal Year: 2024, Volume and Issue: 22(1)

Published: Oct. 2, 2024

Language: Английский

Citations

0
Basement membrane hydrogels dampen CAR-T cell activation: nanofibrillar cellulose gels as alternative to preserve T cell function in 3D cell cultures. DOI Open Access

Sonia Aristín Revilla,

Alessandro Cutilli,

Dedeke Rockx-Brouwer

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 3, 2024

Abstract Background Hydrogel-based 3D culture systems are emerging as a valuable tool for preclinical screening of cell-based immunotherapies against solid and hematological malignancies, such chimeric antigen receptor T (CAR-T) cells. Hydrogels can influence cell function in non-desired manner due to their mechanical properties chemical composition, potentially skewing results testing novel immunotherapeutic compounds. Methods In this study, we assess CD4 + CAR-T activation proliferation chemically-undefined matrices (Matrigel basement membrane extract, BME) compare them synthetic nanofibrillar cellulose (NFC) hydrogel. Results Rheometric analyses show that NFC is more rigid than Matrigel BME. Murine cells acquire regulatory (Treg) phenotype BME, while not observed NFC. Proliferation human higher or Similarly, significantly impaired contrast Conclusions Our findings highlight the impact hydrogel choice on (CAR-)T behavior, with direct implications immunotherapy testing. offers chemically-defined environment where preserved. Key messages What already known topic (preclinical) tumor-killing assays evaluating engineered cytotoxicity, surrounding matrix immune function, activity. Basement hydrogels extract (BME), widely used scaffolds culture, inherently heterogeneous contain extracellular components lymphocyte function. study adds Here, reduced BME compared standard (2D) conditions. contrast, activity preserved gels. Importantly, murine spontaneously Treg cytokine secretion >10-fold lower survival expansion 10-fold How might affect research, practice policy We report intrinsic cytotoxic proliferative potential be underestimated when performing cultures based As an alternative, suggest use chemically defined gels, suitable preserving activation.

Language: Английский

Citations

0