Basement Membrane Invasion Assays: Native Basement Membrane and Chemoinvasion Assay

Schoumacher, Marie; Glentis, Alexandros; Gurchenkov, Vasily; Vignjević, Danijela

doi:10.1007/978-1-62703-538-5_8

Cited by 19 publications

(16 citation statements)

References 12 publications

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“…The transient nature of carcinoma in situ makes the onset of cell invasion and BM breakage laborious to study in vivo. We therefore developed a 3D in vitro model that recapitulates the complexity of carcinoma in situ 19 . We isolated human fibroblasts from primary colon tumor specimens of eight patients (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Cancer-associated fibroblasts induce metalloprotease-independent cancer cell invasion of the basement membrane

et al. 2017

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At the stage of carcinoma in situ, the basement membrane (BM) segregates tumor cells from the stroma. This barrier must be breached to allow dissemination of the tumor cells to adjacent tissues. Cancer cells can perforate the BM using proteolysis; however, whether stromal cells play a role in this process remains unknown. Here we show that an abundant stromal cell population, cancer-associated fibroblasts (CAFs), promote cancer cell invasion through the BM. CAFs facilitate the breaching of the BM in a matrix metalloproteinase-independent manner. Instead, CAFs pull, stretch, and soften the BM leading to the formation of gaps through which cancer cells can migrate. By exerting contractile forces, CAFs alter the organization and the physical properties of the BM, making it permissive for cancer cell invasion. Blocking the ability of stromal cells to exert mechanical forces on the BM could therefore represent a new therapeutic strategy against aggressive tumors.

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Section: Resultsmentioning

confidence: 99%

Cancer-associated fibroblasts induce metalloprotease-independent cancer cell invasion of the basement membrane

et al. 2017

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“…Mice were kept in the Specific Pathogen Free (SPF) animal house of Institut Curie for breeding. Mesentery was isolated from a two-months old female C57BL/6 mouse according to previously described protocol 76,77 . Prior to mesentery isolation, the polyester membrane of a 6.4 mm diameter transwell insert (Corning) was removed using a scalpel.…”

Section: Methodsmentioning

confidence: 99%

Adhesion to nanofibers drives cell membrane remodeling through 1D wetting

Charles-Orszag

Tsai

Bonazzi

et al. 2018

Preprint

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28The shape of cellular membranes is highly regulated by a set of conserved mechanisms. These 29 mechanisms can be manipulated by bacterial pathogens to infect cells. Human endothelial cell 30 plasma membrane remodeling by the bacterium Neisseria meningitidis is thought to be 31 essential during the blood phase of meningococcal infection, but the underlying mechanisms 32 are unknown. Here we show that plasma membrane remodeling occurs independently of F-33 actin, along meningococcal type IV pili fibers, by a novel physical mechanism we term "one-34 dimensional" membrane wetting. We provide a theoretical model that gives the physical basis 35 of 1D wetting and show that this mechanism occurs in model membranes interacting with 36 model nanofibers, and in human cells interacting with model extracellular matrices. It is thus 37 a new general principle driving the interaction of cells with their environment at the nanoscale 38 that is diverted by meningococcus during infection. 39Main text 40 Control of the shape of biological membranes is fundamental for the maintenance of 41 multiple functions in the eukaryotic cell 1 . Acting as the interface of the cell with its 42 surrounding environment, the plasma membrane is a particularly important compartment that 43 is subject to a precise control of its shape and dynamics. Plasma membrane remodeling occurs 44 at very small scales, for example in the biogenesis of caveolae 2 or during the formation of 45 clathrin coated pits 3 . At larger scales, remodeling of the plasma membrane plays an important 46 role in a wide variety of biological processes, such as the uptake of large particles by 47 phagocytosis 4 or in the formation of actin-based membrane structures that support cell 48 migration and probing of the extracellular environment, such as filopodia or lamellipodia 5 . In 49 the context of pathological conditions, especially in bacterial, viral and fungal infections, 50 pathogens manipulate the shape of the plasma membrane to enter host cells. This is often 51 achieved by diverting the actin cytoskeleton 6-8 . Other pathogens remain extracellular and must 52 then resist mechanical strains such as those generated by flow 9 . The bacterium Neisseria 53 meningitidis (or meningococcus) is a human pathogen that, while remaining extracellular 10 , 54 massively remodels the host cell plasma membrane to form filopodia-like protrusions that 55 intercalate between aggregated bacteria upon adhesion to the host cell surface. It was shown 56 in vitro that plasma membrane remodeling allows N. meningitidis to proliferate on the outside 57 of the host cell while mechanically resisting high shear stress levels 11 , suggesting a central role 58 for plasma membrane remodeling in the blood phase of N. meningitidis pathogenesis where 59 bacteria are subject to high shear. Colonization of the blood vessels by N. meningitidis 60 eventually leads to a loss of vascular function that translates into hemorrhagic lesions in 61 organs throughout the body, including the skin where it presents...

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“…Since Chen’s initial description, invadopodia have been observed in many metastatic cancer cell lines (Hoshino et al, 2013) and emerged as one of the key subcellular structures that invasive cells use to breach basement membrane barriers (Lohmer et al, 2014; Schoumacher et al, 2013; Schoumacher et al, 2010). Invadopodia also appear to mediate invasion through the more porous type I collagen rich interstitial matrices that reside between cells and tissues.…”

Section: Background: Invadopodia Are Specialized Subcellular Structurmentioning

confidence: 99%

A new front in cell invasion: The invadopodial membrane

Hastie

Sherwood

2016

European Journal of Cell Biology

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Invadopodia are F-actin-rich membrane protrusions that breach basement membrane barriers during cell invasion. Since their discovery more than 30 years ago, invadopodia have been extensively studied in cancer cells in vitro, where great advances in understanding their composition, formation, cytoskeletal regulation, and control of the matrix metalloproteinase MT1-MMP trafficking have been made. In contrast, few studies examining invadopodia have been conducted in vivo, leaving their physiological regulation unclear. Recent live-cell imaging and gene perturbation studies in C. elegans have revealed that invadopodia are formed with a unique invadopodial membrane, defined by its specialized lipid and associated protein composition, which is rapidly recycled through the endolysosome. Here, we provide evidence that the invadopodial membrane is conserved and discuss its possible functions in traversing basement membrane barriers. Discovery and examination of the invadopodial membrane has important implications in understanding the regulation, assembly, and function of invadopodia in both normal and disease settings.

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Basement Membrane Invasion Assays: Native Basement Membrane and Chemoinvasion Assay

Cited by 19 publications

References 12 publications

Cancer-associated fibroblasts induce metalloprotease-independent cancer cell invasion of the basement membrane

Cancer-associated fibroblasts induce metalloprotease-independent cancer cell invasion of the basement membrane

Adhesion to nanofibers drives cell membrane remodeling through 1D wetting

A new front in cell invasion: The invadopodial membrane

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