3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs

Cancers, Journal Year: 2021, Volume and Issue: 14(1), P. 190 - 190

Published: Dec. 31, 2021

Today, innovative three-dimensional (3D) cell culture models have been proposed as viable and biomimetic alternatives for initial drug screening, allowing the improvement of efficiency development. These are gaining popularity, given their ability to reproduce key aspects tumor microenvironment, concerning 3D architecture well interactions cells with extracellular matrix surrounding non-tumor cells. The development accurate may become beneficial decrease use laboratory animals in scientific research, accordance European Union's regulation on 3R rule (Replacement, Reduction, Refinement). This review focuses impact cancer discussing advantages, limitations, compatibility high-throughput screenings automated systems. An insight is also adequacy available readouts interpretation data obtained from models. Importantly, we emphasize need incorporation additional complementary microenvironment elements design models, towards improved predictive value efficacy.

Language: Английский

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Cell–3D matrix interactions: recent advances and opportunities

Trends in Cell Biology, Journal Year: 2022, Volume and Issue: 32(10), P. 883 - 895

Published: April 8, 2022

The diversity of hundreds extracellular matrix (ECM) molecules in different tissues and their interactions are now being documented 'matrisome' databases.Physical properties the 3D ECM, including viscoelasticity microarchitecture, can govern cell adhesion, mechanotransduction, multiple modes migration.New advances ECM biology identifying mechanisms cancer progression fibrosis, as well potential therapeutic targets.Characterizations cell–ECM feedback loops computational modeling providing new insights opportunities for intervention diseases disorders. Tissues consist cells surrounding (ECM). Cell–ECM play crucial roles embryonic development, differentiation, tissue remodeling, fibrosis cancer. Recent research characterizing cell–matrix include detailed descriptions associated molecules, complex intermolecular development disease, identification distinctive migration ECMs, into organ formation. Exploring physical features microenvironments bidirectional regulation signaling organization emphasize dynamic nature these interactions, which that exacerbate disease. Understanding potentially lead to targeted interventions. New on interactionsThe with (see Glossary) during formation adult homeostasis, pathogenesis such This field has expanded explosively after discovery many surface receptors. Our goal this brief review is highlight recent conceptual experimental should provide exciting future cell–3D interactions.Diversity interactionsA starting ~2019 been widespread adoption term – is, comprising it changes disease pathogenesis. holistic concept matrisomes moves beyond classical studies focusing a single protein or family not only structural proteins, collagens, elastin, proteoglycans, fibronectin, but also matrix-associated enzymes inhibitors, matrix-bound growth factors, some cases receptors [1.Karamanos N.K. et al.A guide composition functions matrix.FEBS J. 2021; 288: 6850-6912Crossref PubMed Scopus (65) Google Scholar,2.Izzi V. al.Pan-cancer analysis genomic alterations mutations matrisome.Cancers (Basel). 2020; 12: 2046Crossref (27) Scholar]. Among examples, have used matrisome analyses characterize basement membranes (https://bmbase.manchester.ac.uk/), discover ECM-associated genes more than other [2.Izzi Scholar], identify thrombospondin tenascin links collagen alignment breast [3.Tomko L.A. al.Targeted identifies thrombospondin-2 tenascin-C aligned stroma from invasive carcinoma.Sci. Rep. 2018; 8: 12941Crossref (37) bioengineered models human pancreatic [4.Osuna de la Pena D. al.Bioengineered recapitulate vivo tumour biology.Nat. Commun. 5623Crossref (11) Scholar].Many publications still at descriptive level. There considerable overlaps between components 'adhesome,' comprises adhesions, especially focal adhesions (e.g., see www.adhesome.org). Both 'omics' approaches major applying increasingly sophisticated methods understand involving networks rather just few selected proteins past. Unexpected findings may arise terms groups regulating components. Exemplifying crosstalk, planar membrane two its biochemically unrelated constituents, laminin IV, strongly regulate assembly fibrillar component, variety types [5.Lu al.Basement regulates fibronectin using sliding driven by contractile winch.Dev. Cell. 52: 631-646 e634Abstract Full Text PDF (24) Scholar].We know vary widely depending type biochemical mechanical (Figure 1). Multiple characterized recently range lamellipodial characteristic mesenchymal fibroblast-like rounded amoeboid immune certain [6.Bodor D.L. al.Of shapes motion: basis animal migration.Dev. 550-562Abstract (45) Scholar,7.Yamada K.M. Sixt M. Mechanisms migration.Nat. Rev. Mol. Cell Biol. 2019; 20: 738-752Crossref (262) lobopodial cross-linked linearly elastic spatially confined intracellular pressure, cortical actin flow, ion fluxes, [8.Zhao R. al.Cell sensing decision-making confinement: role TRPM7 tug war hydraulic pressure cross-sectional area.Sci. Adv. 5eaaw7243Crossref (35) Scholar, 9.Patel S. al.Myosin II Arp2/3 cross-talk governs lamellipodia formation.Mol. 32: 579-589Crossref 10.Ullo M.F. Logue J.S. ADF cofilin-1 collaborate promote flow leader bleb-based cells.eLife. 10e67856Crossref (6) 11.Reversat A. al.Cellular locomotion environmental topography.Nature. 582: 582-585Crossref (69) 12.Yolland L. al.Persistent polarized global essential directionality 21: 1370-1381Crossref (22) Scholar].The microarchitecture affect differentiation [13.Doyle A.D. al.Local microenvironment through spatiotemporal dynamics contractility-dependent adhesions.Nat. 2015; 6: 8720Crossref (268) Scholar,14.Seo B.R. al.Collagen mechanically controls myofibroblast differentiation.Proc. Natl. Acad. Sci. U. 117: 11387-11398Crossref (58) For example, networks, fiber thickness pore size, adipose stromal toward process independent overall stiffness previously known stem [14.Seo Scholar,15.Chaudhuri O. al.Effects cellular behaviour.Nature. 584: 535-546Crossref (427) Scholar].Matrix propertiesThe local migratory speed show differences molecular elasticity [15.Chaudhuri Examining whether an soft stiff numerous previous [16.Hayward M.K. al.Tissue mechanics fate, cancer.Dev. 56: 1833-1847Abstract (20) Scholar,17.Xue B. al.Engineering hydrogels homogeneous controlling lineage specification.Proc. 118e2110961118Crossref (13) Scholar] ideally be complemented behavior environments differing viscoelasticity. reason biological matrices often viscoelastic, they display combination viscosity thick fluid attempts return material original form deforming force released. results plastic deformation slipping, creep 'stress relaxation' response deformed, without returning form. Viscoelasticity modulated extent crosslinking property effects behavior, although underlying yet clear. altered rheology filopodial versus protrusions leading edge cell, rates spreading migration, processes morphogenesis, epithelial-mesenchymal transition, invasion, [18.Adebowale K. al.Enhanced substrate stress relaxation promotes filopodia-mediated Mater. 1290-1299Crossref (36) 19.Gong Z. al.Matching timescales maximizes viscoelastic substrates.Proc. 115: E2686-E2695Crossref (129) 20.Wisdom al.Matrix plasticity confining microenvironments.Nat. 9: 4144Crossref (162) 21.Yang mechanosensing synthetic controlled biophysical dynamics.Nat. 3514Crossref 22.Indana al.Viscoelasticity adhesion biomaterials control pluripotent morphogenesis culture.Adv. 33e2101966Crossref (17) 23.Chang A.C. al.Precise tuning characterization interfaces study early transition behaviors.Langmuir. 2022; (Published online February 10, 2022)https://doi.org/10.1021/acs.langmuir.1c03048Crossref (3) 24.Hui E. al.The combined influence adhesive cues fibroblast organization.Cell. Bioeng. 14: 427-440Crossref (9) fact chemical substantially alter mode, speed, means one cannot simply 'work 3D' due factors An ongoing challenge will generate accurately mimic specific use ex tissues. Development physiological simple gels current valuable accurate platforms testing translational approaches.MechanotransductionDifferent elicit distinct responses cells. 2D revealed test repetitively probing [25.Plotnikov S.V. al.Force fluctuations within mediate ECM-rigidity directed migration.Cell. 2012; 151: 1513-1527Abstract (559) analogous hikers footing when crossing unstable terrain. Cells sense respond ECM-transmitted forces tension involves integrin-based where actomyosin-mediated transmitted substrata [26.Zuidema al.Crosstalk complexes mechanotransduction.Bioessays. 42e2000119Crossref (31) Scholar,27.Doyle al.Cell-extracellular dynamics.Phys. 19https://doi.org/10.1088/1478-3975/ac4390Crossref (8) In environments, mechanotransduction level resembles under conditions stiffer substrates stabilize while softer, flexible shorter lifetimes faster Scholar].Mechanotransduction becomes particularly concerning cycle. evidence fibroblasts migrating reveals prior translocation, initially deform (prestrain) fibrils, increasing self-generated contracting transmitting essentially first 'pulling up slack rope' (i.e., fibrils [28.Doyle al.3D anterior contraction generates prestrain.Dev. 826-841.e4Abstract Scholar]). Interestingly, epithelial relatively equal-and-opposite strain transmission posterior directions 2A ) [29.Hall M.S. al.Fibrous nonlinear enables positive ECMs.Proc. 2016; 113: 14043-14048Crossref (174) (nonepithelial) cancers constant prestrains twofold greater front rear suggesting disconnect propagation 2A) prestrain likely genetically primed contractility-centric mode higher expression myosin II) enhanced microenvironment. sequence events distinctive, actomyosin contractions preceding leading-edge protrusive activity, helps establish unique cycle 2B–C) Discrepancies cycles importance conditions.Figure 2Mechanotransduction migration.Show full caption(A) Schematic showing directionally gels. Mesenchymal (left) high extensive integrin ligation large strains larger rear. Similar attributes seen fibrosarcoma cells, majority (right) smaller, transient rear, lower graph summarizing latter concept, requires further testing). (B C) schematic shows matrix. Yellow arrows depict directional applied matrix, magenta indicate relative summed given region. During cycle, (C) retrograde pull stabilizes (gray ovals) edge. A contralateral anterograde (increased direction migration) leads pinching followed increase protrusion (broken white line). Abbreviation: matrix.View Large Image Figure ViewerDownload Hi-res image Download (PPT)In addition, (elastic etc.) alters mechanotransduction. suggest ECMs Viscoelastic effective eliciting compared Future evaluate interacting properties, vitro microenvironments.The nucleus migrationSome 'nuclear piston' pulled anteriorly contractility pressurize drive 'lobopodial' forward [30.Petrie R.J. al.Activating nuclear piston mechanism tumor cells.J. 2017; 216: 93-100Crossref (63) (plastic), mechanosensitive channels generated elevated hydrostatic triggering channel activation; resulting influx sodium calcium ions enhances osmotic provides additional extending promoting efficient [31.Lee H.P. activates paths microenvironments.Sci. 7eabd4058Google Scholar].Another intriguing finding stiff, bulky ruler help choice wider, readily traversed passageway [32.Renkawitz al.Nuclear positioning facilitates along path least resistance.Nature. 568: 546-550Crossref (125) Scholar,33.Lomakin A.J. acts tailoring spatial constraints.Science. 370eaba2894Crossref (110) Besides serving ruler, conjunction cytoskeleton function, gauge activate epigenetic pathways 3) Scholar,34.Venturini measures shape proprioception behavior.Science. 370eaba2644Crossref (102) 35.Maurer Lammerding driving force: disease.Annu. Biomed. Eng. 443-468Crossref (80) 36.Alisafaei F. al.Regulation architecture, mechanics, nucleocytoplasmic shuttling geometric constraints.Proc. 116: 13200-13209Crossref (82) Scholar].Figure 3Multiple mechanosensing, mechanotransduction.Show captionThe nucleus, largest organelle, function entry narrow spaces channels. contraction, thereby termed lobopodia. serve sensor responding confinement. Finally, signal transducer initiating gene expression.View (PPT)Cancer dynamicsCancer invasion continues another very active investigation various ECM. Before malignant invade interstitial tissues, must usually breach barrier surrounds 4). Although proteases degrade protease-independent breaching occur. Physical extension penetrate expand holes ATP production Caenorhabditis elegans [37.Kelley L.C. al.Adaptive F-actin polymerization localized absence MMPs.Dev. 48: 313-328.e8Abstract Human nonproteolytic repetitive microspikes widen filopodia enlarging perforations; protruding subsequently probe [38.Chang Chaudhuri Beyond proteases: invasion.J. 218: 2456-2469Crossref 39.Eschenbruch al.From fibers: remodeling acini drives II-mediated invasion.Cells. 10: 1979Crossref 40.Gong al.Recursive dissipation chemo-mechanical oscillatory invadopodia.Cell 35109047Abstract Cellular metabolic activation important successful crosstalk Scholar,41.Zanotelli M.R. al.Mechanoresponsive metabolism metastasis.Cell Metab. 33: 1307-1321Abstract 42.Romani P. metabolism.Nat. 22: 22-38Crossref (94) 43.Torrino al.Mechano-induced microtubule glutamylation 1342-1357.e10Abstract (18) 4Different membrane.Show captionCancer transiently barriers mechanisms. proteases, metalloproteinases (MMPs), locally, MMPs tips invadopodia. Even if inhibited, tiny perforations requiring energy (orange shading) (yellow arrows) push laterally ridge around expanding hole (red mounds). both processes, proteolysis perforations.View (PPT)Although generation thought involve contractility, relationship levels nonmuscle invasiveness complex. Some isoforms actomyosin-associated predicted facilitate others decreased consistent suppressor Scholar,44.Parajon mechanobiome: goldmine therapeutics.Am. Physiol. 320: C306-C323Crossref Scholar,45.Picariello H.S. IIA suppresses glioblastoma sensitive manner.Proc. 15550-15559Crossref (25) bifunctional complexity myosin-X enhancing suppressing modulating [46.Peuhu al.Myosin-X-dependent limits invasion.bioRxiv. October 22, 2021)https://doi.org/10.1101/2021.10.22.464987Google Scholar].Although obvious source disrupting structure breaching, contributor lateral branches density [47.Papalazarou Maches

Language: Английский

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Synthetic extracellular matrices with function-encoding peptides

Nature Reviews Bioengineering, Journal Year: 2023, Volume and Issue: 1(7), P. 518 - 536

Published: April 25, 2023

Language: Английский

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Patient-specific modeling of stroma-mediated chemoresistance of pancreatic cancer using a three-dimensional organoid-fibroblast co-culture system

Journal of Experimental & Clinical Cancer Research, Journal Year: 2022, Volume and Issue: 41(1)

Published: Oct. 22, 2022

Cancer-associated fibroblasts (CAFs) are considered to play a fundamental role in pancreatic ductal adenocarcinoma (PDAC) progression and chemoresistance. Patient-derived organoids have demonstrated great potential as tumor avatars for drug response prediction PDAC, yet they disregard the influence of stromal components on chemosensitivity.We established direct three-dimensional (3D) co-cultures primary PDAC patient-matched CAFs investigate effect fibroblastic compartment sensitivity gemcitabine, 5-fluorouracil paclitaxel treatments using an image-based assay. Single-cell RNA sequencing was performed three organoid/CAF pairs mono- co-culture uncover transcriptional changes induced by tumor-stroma interaction.Upon with CAFs, we observed increased proliferation reduced chemotherapy-induced cell death organoids. data evidenced induction pro-inflammatory phenotype co-cultures. Organoids showed expression genes associated epithelial-to-mesenchymal transition (EMT) several receptor-ligand interactions related EMT were identified, supporting key CAF-driven chemoresistance.Our results demonstrate personalized models not only profiling but also unraveling molecular mechanisms involved chemoresistance-supporting stroma.

Language: Английский

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3D Biomimetic Models to Reconstitute Tumor Microenvironment In Vitro: Spheroids, Organoids, and Tumor‐on‐a‐Chip

Advanced Healthcare Materials, Journal Year: 2023, Volume and Issue: 12(18)

Published: March 14, 2023

Abstract Decades of efforts in engineering vitro cancer models have advanced drug discovery and the insight into biology. However, establishment preclinical that enable fully recapitulating tumor microenvironment remains challenging owing to its intrinsic complexity. Recent progress techniques has allowed development a new generation can recreate complex vivo microenvironments accurately predict responses, including spheroids, organoids, tumor‐on‐a‐chip. These biomimetic 3D are particular interest as they pave way for better understanding biology accelerating anticancer therapeutics with reducing animal use. Here, recent advances developing these platforms modeling screening, focusing on incorporating hydrogels reviewed reconstitute physiologically relevant microenvironments. The combination spheroids/organoids microfluidic technologies is also highlighted mimic tumors discuss challenges future directions clinical translation such screening personalized medicine.

Language: Английский

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Drug screening on digital microfluidics for cancer precision medicine

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: May 22, 2024

Abstract Drug screening based on in-vitro primary tumor cell culture has demonstrated potential in personalized cancer diagnosis. However, the limited number of cells, especially from patients with early stage cancer, hindered widespread application this technique. Hence, we developed a digital microfluidic system for drug using cells and established working protocol precision medicine. Smart control logic was to increase throughput decrease its footprint parallelly screen three drugs 4 × cm 2 chip device measuring 23 16 3.5 3 . We validated method an MDA-MB-231 breast xenograft mouse model liver specimens patients, demonstrating suppression mice/patients treated that were screened be effective individual cells. Mice on-chip as ineffective exhibited similar results those groups. The identified through consistency absence mutations their related genes determined via exome sequencing tumors, further validating protocol. Therefore, technique may promote advances medicine treatment and, eventually, any disease.

Language: Английский

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A patient-specific lung cancer assembloid model with heterogeneous tumor microenvironments

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: April 20, 2024

Abstract Cancer models play critical roles in basic cancer research and precision medicine. However, current vitro are limited by their inability to mimic the three-dimensional architecture heterogeneous tumor microenvironments (TME) of vivo tumors. Here, we develop an innovative patient-specific lung assembloid (LCA) model using droplet microfluidic technology based on a microinjection strategy. This method enables precise manipulation clinical microsamples rapid generation LCAs with good intra-batch consistency size cell composition evenly encapsulating patient tumor-derived TME cells organoids inside microgels. recapitulate inter- intratumoral heterogeneity, cellular diversity, genomic transcriptomic landscape parental LCA could reconstruct functional heterogeneity cancer-associated fibroblasts reflect influence drug responses compared organoids. Notably, accurately replicate outcomes patients, suggesting potential predict personalized treatments. Collectively, our studies provide valuable for precisely fabricating assembloids promising

Language: Английский

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Heterotrophic bacterial diazotrophs are more abundant than their cyanobacterial counterparts in metagenomes covering most of the sunlit ocean

The ISME Journal, Journal Year: 2021, Volume and Issue: 16(4), P. 927 - 936

Published: Oct. 25, 2021

Language: Английский

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Programmable DNA Hydrogels as Artificial Extracellular Matrix

Small, Journal Year: 2022, Volume and Issue: 18(36)

Published: Feb. 4, 2022

Abstract The cell microenvironment plays a crucial role in regulating behavior and fate physiological pathological processes. As the fundamental component of microenvironment, extracellular matrix (ECM) typically possesses complex ordered structures provides essential physical chemical cues to cells. Hydrogels have attracted much attention recapitulating ECM. Compared natural synthetic polymer hydrogels, DNA hydrogels unique programmable capability, which endows material precise structural customization tunable properties. This review focuses on recent advances as artificial matrix, particularly pure hydrogels. It introduces classification, design, assembly then summarizes state‐of‐the‐art achievements encapsulation, culture, tissue engineering with Ultimately, challenges prospects for cellular applications are delivered.

Language: Английский

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Biomaterial-based platforms for tumour tissue engineering

Nature Reviews Materials, Journal Year: 2023, Volume and Issue: 8(5), P. 314 - 330

Published: Feb. 14, 2023

Language: Английский

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