Modes Of Invasion Research Articles

Marmoset and human trophoblast stem cells differ in signaling requirements and recapitulate divergent modes of trophoblast invasion

Early human trophoblast development has remained elusive due to the inaccessibility of the early conceptus. Non-human primate models recapitulate many features of human development and allow access to early postimplantation stages. Here, we tracked the pre- to postimplantation transition of the trophoblast lineage in superficially implanting marmoset embryos invivo. We differentiated marmoset naive pluripotent stem cells into trophoblast stem cells (TSCs), which exhibited trophoblast-specific transcriptome, methylome, differentiation potential, and long-term self-renewal. Notably, human TSC culture conditions failed to support marmoset TSC derivation, instead inducing an extraembryonic mesoderm-like fate in marmoset cells. We show that combined MEK, TGF-β/NODAL, and histone deacetylase inhibition stabilizes a periimplantation trophoblast-like identity in marmoset TSCs. By contrast, these conditions differentiated human TSCs toward extravillous trophoblasts. Our work presents a paradigm to harness the evolutionary divergence in implantation strategies to elucidate human trophoblast development and invasion.

Live-Cell Invasive Phenotyping Uncovers ALK2 as a Therapeutic Target in LKB1-Mutant Lung Cancer.

The acquisition of invasive properties is a prerequisite for tumor progression and metastasis. Molecular subtypes of KRAS-driven lung cancer exhibit distinct modes of invasion that contribute to unique growth properties and therapeutic susceptibilities. Despite this, pre-clinical strategies designed to exploit growth within the context of invasion are lacking. To address this, we designed an experimental system to screen for targetable signaling pathways linked to active early 3D invasion phenotypes in different molecular subtypes of KRAS-driven lung adenocarcinoma (LUAD). Combined live-cell imaging of human bronchial epithelial cells in a 3D invasion matrix and transcriptomic profiling identified mutant LKB1-specific upregulation of BMP6. LKB1 loss increased BMP6 signaling, which induced the canonical iron regulatory hormone hepcidin. Intact LKB1 was necessary to maintain BMP6 signaling homeostasis and restrict ALK2/BMP6-fueled growth. Pre-clinical studies in a Kras/Lkb1-mutant syngeneic mouse model and in a xenograft model showed potent growth suppression by inhibiting the ALK2/BMP6 signaling axis with single agent inhibitors that are currently in clinical trials. Lastly, BMP6 expression was elevated in LKB1-mutant early-stage lung cancer patient tumors. These results are consistent with a model where LKB1 acts as a 'brake' to iron regulated growth and suggest that ALK2 inhibition can be used for patients with LKB1-mutant tumors.

DIPG-27. MATCHED PRIMARY-RELAPSE PEDHGG PATIENT-DERIVED XENOGRAFT MODELS TO IDENTIFY THERAPY RESISTANCE PROFILES

Abstract BACKGROUND We established patient-derived xenografts (PDXs) from matched primary-relapse pairs of pediatric high-grade gliomas (pedHGGs) in collaboration with the Swedish Childhood Tumor Biobank. Further, we used human iPSC derived cortical organoids to evaluate interactions between glial and tumor cells under treatment pressure potentially lost in xenograft models. Using this setup, we hope to follow tumor relapse evolution and to identify novel mechanisms behind treatment evasion and relapse formation. METHODS Tumoral singel cell suspensions from matched primary-relapse pairs are injected orthotopically in immunodeficient NOG mice. We establish in vitro 2D models in stem cell conditions from the first generation of animals transplanted. Lentivirus-based labelling with GFP/Luciferase was done on PDX-derived cell lines to allow cell tracking and to quantify efficacy of drug screening efforts. Co-culture of these cells with normal cells in 3D assembloids were performed using 150-250 days old cortical organoids. Single-cell RNA sequencing and bulk RNA sequencing (RNA-seq) on patient biopsies, their corresponding PDXs from first generation of mice, and PDX-derived cell lines, was performed to track potential changes in vivo and in vitro and to further validate primary-relapse specific evolution. RESULTS 2D and 3D cell cultures offered efficient labeling and we quantified differences in sensitivity to standard therapy between primary and relapse samples. PDX models and assembloids highlighted pedHGG growth patterns, modes of invasion and cell interactions, especially astrocyte-tumor cell interactions. These 3D models also allow for further validation of targeted drug testing based on multiomics data (WGS, methylation and RNA-seq) from the patient biopsies. CONCLUSION PedHGG primary-relapse in vitro 2D cell line/ in vivo 3D assembloid models together with PDXs is a robust approach to study different aspects of relapse-specific tumor biology. Our results offer insights aiding the development of novel therapeutics inhibiting relapse mechanisms already in the first line of treatment.

Subpopulation commensalism promotes Rac1-dependent invasion of single cells via laminin-332.

Phenotypic heterogeneity poses a significant hurdle for cancer treatment but is under-characterized in the context of tumor invasion. Amidst the range of phenotypic heterogeneity across solid tumor types, collectively invading cells and single cells have been extensively characterized as independent modes of invasion, but their intercellular interactions have rarely been explored. Here, we isolated collectively invading cells and single cells from the heterogeneous 4T1 cell line and observed extensive transcriptional and epigenetic diversity across these subpopulations. By integrating these datasets, we identified laminin-332 as a protein complex exclusively secreted by collectively invading cells. Live-cell imaging revealed that laminin-332 derived from collectively invading cells increased the velocity and directionality of single cells. Despite collectively invading and single cells having similar expression of the integrin α6β4 dimer, single cells demonstrated higher Rac1 activation upon laminin-332 binding to integrin α6β4. This mechanism suggests a novel commensal relationship between collectively invading and single cells, wherein collectively invading cells promote the invasive potential of single cells through a laminin-332/Rac1 axis.

N- and s-substituted Pyrazolopyrimidines: A promising new class of potent c-Src kinase inhibitors with prominent antitumor activity

In this work, readily achievable synthetic pathways were utilized for construction of a library of N/S analogues based on the pyrazolopyrimidine scaffold with terminal alkyl or aryl fragments. Subsequently, we evaluated the anticancer effects of these novel analogs against the proliferation of various cancer cell lines, including breast, colon, and liver lines. The results were striking, most of the tested molecules exhibited strong and selective cytotoxic activity against the MDA-MB-231 cancer cell line; IC50 1.13 µM. Structure-activity relationship (SAR) analysis revealed that N-substituted derivatives generally enhanced the cytotoxic effect, particularly with aliphatic side chains that facilitated favorable target interactions. We also investigated apoptosis, DNA fragmentation, invasion assay, and anti-migration effects, and discussed their underlying molecular mechanisms for the most active compound 7c. We demonstrated that 7c N-propyl analogue could inhibit MDA-MB-231 TNBC cell proliferation by inducing apoptosis through the regulation of vital proteins, namely c-Src, p53, and Bax. In addition, our results also revealed the potential of these compounds against tumor metastasis by downregulating the invasion and migration modes. Moreover, the in vitro inhibitory effect of active analogs against c-Src kinase was studied and proved that might be the main cause of their antiproliferative effect. Overall, these compelling results point towards the therapeutic potential of these derivatives, particularly those with N-substitution as promising candidates for the treatment of TNBC type of breast cancer.

Multiscale model of the different modes of cancer cell invasion.

Mathematical models of biological processes altered in cancer are built using the knowledge of complex networks of signaling pathways, detailing the molecular regulations inside different cell types, such as tumor cells, immune and other stromal cells. If these models mainly focus on intracellular information, they often omit a description of the spatial organization among cells and their interactions, and with the tumoral microenvironment. We present here a model of tumor cell invasion simulated with PhysiBoSS, a multiscale framework, which combines agent-based modeling and continuous time Markov processes applied on Boolean network models. With this model, we aim to study the different modes of cell migration and to predict means to block it by considering not only spatial information obtained from the agent-based simulation but also intracellular regulation obtained from the Boolean model. https://github.com/sysbio-curie/Invasion_model_PhysiBoSS.

Plasticity of cancer invasion and energy metabolism.

Energy deprivation is a frequent adverse event in tumors that is caused by mutations, malperfusion, hypoxia, and nutrition deficit. The resulting bioenergetic stress leads to signaling and metabolic adaptation responses in tumor cells, secures survival, and adjusts migration activity. The kinetic responses of cancer cells to energy deficit were recently identified, including a switch of invasive cancer cells to energy-conservative amoeboid migration and an enhanced capability for distant metastasis. We review the energy programs employed by different cancer invasion modes including collective, mesenchymal, and amoeboid migration, as well as their interconversion in response to energy deprivation, and we discuss the consequences for metastatic escape. Understanding the energy requirements of amoeboid and other dissemination strategies offers rationales for improving therapeutic targeting of metastatic cancer progression.

ARF3 weights the balance for prostate cancer metastasis.

Patrick T. Caswell discusses work from Bryant and colleagues (2023. J. Cell Biol.https://doi.org/10.1083/jcb.202206115) which shows how ARF3 impacts metastasis in prostate cancer by regulating a switch between modes of collective invasion.

Inhibition of MMPs supports amoeboid angiogenesis hampering VEGF-targeted therapies via MLC and ERK 1/2 signaling

BackgroundIn the past decades studies on anti-tumoral drugs inhibiting matrix metalloproteinase (MMPs) were disappointing. Recently, we demonstrated that mature endothelial cells (ECs) and endothelial colony forming cells (ECFCs) can switch between invasion modes to cope with challenging environments, performing the “amoeboid angiogenesis” in the absence of proteases activity.MethodsWe first set out to investigate by ELISA if the inhibitors of the main protease family involved in angiogenesis were differently expressed during breast cancer progression. We used Marimastat, a broad-spectrum MMP inhibitor, as a means of inducing amoeboid characteristics and studied VEGF role in amoeboid angiogenesis. Thus, we performed invasion and capillary morphogenesis assay, morphological, cell signaling and in vivo mouse studies.ResultsOur data showed that TIMP1, TIMP2, alpha2-antiplasmin, PAI-1 and cystatin increase in breast cancer serum of patients with primary cancer and lymph node positive compared to healthy women. In vitro results revealed that the most high-powered protease inhibitors able to induce amoeboid invasion of ECFCs were TIMP1, 2 and 3. Surprisingly, Marimastat promotes ECFC invasion and tubular formation in vitro and in vivo, inducing amoeboid characteristics. We observed that the combination of Marimastat plus VEGF doesn’t boost neither cell invasion nor vessel formation capacity. Moreover, inhibition of VEGF activity with Bevacizumab in the presence of Marimastat confirmed that amoeboid angiogenesis is independent from the stimulus of the main vascular growth factor, VEGF.ConclusionsWe underline the importance to consider the amoeboid mechanism of endothelial and cancer cell invasion, probably responsible for the failure of synthetic metalloproteinase inhibitors as cancer therapy and tumor resistance to VEGF-targeted therapies, to set-up new drugs to be used in cancer therapy.

Crossover from viscous fingering to fracturing in cohesive wet granular media: a photoporomechanics study.

We study fluid-induced deformation and fracture of cohesive granular media, and apply photoporomechanics to uncover the underpinning grain-scale mechanics. We fabricate photoelastic spherical particles of diameter d = 2 mm, and make a monolayer granular pack with tunable intergranular cohesion in a circular Hele-Shaw cell that is initially filled with viscous silicone oil. We inject water into the oil-filled photoelastic granular pack, varying the injection flow rate, defending-fluid viscosity, and intergranular cohesion. We find two different modes of fluid invasion: viscous fingering, and fracturing with leak-off of the injection fluid. We directly visualize the evolving effective stress field through the particles' photoelastic response, and discover a hoop effective stress region behind the water invasion front, where we observe tensile force chains in the circumferential direction. Outside the invasion front, we observe compressive force chains aligning in the radial direction. We conceptualize the system's behavior by means of a two-phase poroelastic continuum model. The model captures granular pack dilation and compaction with the boundary delineated by the invasion front, which explains the observed distinct alignments of the force chains. Finally, we rationalize the crossover from viscous fingering to fracturing by comparing the competing forces behind the process: viscous force from fluid injection that drives fractures, and intergranular cohesion and friction that resist fractures.

Cancer-associated fibroblast-dependent and -independent invasion of gastric cancer cells.

Cancer cells are known to exhibit a cancer-associated fibroblast (CAF)-dependent invasive mode in the presence of CAFs. The purpose of this study was to investigate whether intrinsic factors of gastric cancer cells influence the CAF-dependent invasive mode of cancer cells. We observed dynamic movement of CAFs, and cancer cells, by time-lapse imaging of 2-D and 3-D collagen invasion models, and evaluated invasion modes of gastric cancer cell lines (MKN-7, MKN-45, and HSC44PE). We further examined whether modification of invasive capacity of CAFs can alter invasive mode of MKN-7, and HSC44PE cells. When MKN-7 and MKN-45 cells were co-cultured with CAFs, CAFs first invade collagen matrix followed by cancer cells (CAF-dependent invasion), whereas HSC44PE cells invaded collagen matrix independent of CAFs' invasion. Overexpression or suppression of podoplanin in CAFs, respectively, increased or decreased the invasive capacity of CAFs, and significantly increased or decreased the number of invading MKN-7 cells, respectively. CAFs overexpressing a podoplanin mutant, lacking the cytoplasmic domain, had significantly reduced invasive capacity, compared to CAFs overexpressing wild-type podoplanin, and it also reduced the number of invading MKN-7 cells significantly. When HSC44PE cells, and CAFs were co-cultured, changes in the podoplanin expression in CAFs similarly altered the invasive capacity of CAFs, but it did not affect the number of invading HSC44PE cells. These results indicate that in presence of CAFs, gastric cancer cells exhibit both CAF-dependent and -independent modes of invasion, the determinants of which may depend on the intrinsic properties of the gastric cancer cells.

Direct Prediction of Fluid‐Fluid Displacement Efficiency in Ordered Porous Media Using the Pore Structure

AbstractFluid‐fluid displacement in porous media is common in many natural and engineering settings. Extensive studies investigated the transition of displacement patterns, but the direct prediction of the displacement efficiency using the pore structure is lacking. Here, we propose a method to directly predict the displacement efficiency with no need to solve the Navier‐Stokes and the Hagen‐Poiseuille equations in ordered porous media. The predictive method origins from the pore‐scale filling events, which can be divided into two directions such as the bulk flow direction and the transverse direction. The pore‐filling event (burst) dominates the fluid invasion for the bulk flow direction, and the invading phase forms a thin fingering channel. For the transverse direction, we introduce three invasion modes (compact, taper, and widen) to quantify fluid invasion. We can predict the finger width in each column, and the displacement efficiency can be predicted through the weighted average of the predicted finger width. We evaluate the predictive method using microfluidic experiments and pore‐network simulations, confirming that the predictive method can reasonably predict the displacement efficiency in ordered porous media. Our method can also be applicable for disorder porous media when the disorder is smaller than a critical value. The predictive method can directly predict fluid invasion according to pore structure, thus greatly improving the computational efficiency and is of significance in multiphase flow control.

Tumour invasion and dissemination.

Activating invasion and metastasis are one of the primary hallmarks of cancer, the latter representing the leading cause of death in cancer patients. Whilst many advances in this area have been made in recent years, the process of cancer dissemination and the underlying mechanisms governing invasion are still poorly understood. Cancer cells exhibit multiple invasion strategies, including switching between modes of invasion and plasticity in response to therapies, surgical interventions and environmental stimuli. The ability of cancer cells to switch migratory modes and their inherent plasticity highlights the critical challenge preventing the successful design of cancer and anti-metastatic therapies. This mini-review presents current knowledge on the critical models of tumour invasion and dissemination. We also discuss the current issues surrounding current treatments and arising therapeutic opportunities. We propose that the establishment of novel approaches to study the key biological mechanisms underlying the metastatic cascade is critical in finding novel targets that could ultimately lead to complete inhibition of cancer cell invasion and dissemination.

Water Invasion Mode of Carbonate Gas Reservoirs Controlled by Edge Water: Three Invasion Modes

Water Invasion Mode of Carbonate Gas Reservoirs Controlled by Edge Water: Three Invasion Modes

Research Progress on Spread Through Air Spaces of Lung Cancer

气腔扩散（spread through air spaces, STAS）是2015版世界卫生组织（World Health Organization, WHO）肺癌病理分类中首次提出的概念，是指在主瘤体边界外的气腔内出现微乳头状细胞簇、实性细胞巢或单个肿瘤细胞。除传统观念的肺腺癌浸润方式（间质、脉管及胸膜侵犯）外，STAS被确定为第四种肺浸润性腺癌的侵袭模式。近年来，关于STAS的研究成为热点，STAS的存在不仅与肺癌病理组织学、基因突变等因素相关，诸多研究也证实其可作为肿瘤复发及预后的独立相关因素。但是，也有研究认为人为因素可以造成STAS的形态学假象，需在临床工作中注意甄别。本文就STAS的分类、相关病理学特征、基因状态改变以及可能造成STAS假象的人为因素等方面的研究进展予以综述。

Experimental Study on the Suspension Amount of Invasion Gas in Drilling Fluid with Yield Stress

The gas suspension phenomenon caused by the yield stress of the drilling fluid affects the accurate calculation of wellbore pressure after gas invasion. At present, most studies on the bubble suspension in the yield stress fluid focus on the single-bubble suspension condition and there are few studies on the gas suspension concentration. This paper carried out the GSC (gas suspension concentration) experiment in the simulated drilling fluid, xanthan solution, with different gas invasion methods. The GSC in the drilling fluid under the conditions of diffuse gas invasion and differential pressure gas invasion was simulated by using two methods of stir-depressurization and continuous ventilation. The results showed that when the size of a single bubble satisfied the single-bubble suspension condition, multiple bubbles can be suspended at the same time. The GSC is affected by the average size of the suspended bubbles, the yield stress of the drilling fluid, and the gas invasion modes. For different gas invasion modes, the empirical models of critical GSC related to the dimensionless number Bi are established. Compared with the experimental data, the relative error of the critical GSC in diffuse gas invasion is less than 6% and the relative error of the critical GSC in differential pressure gas invasion is less than 10%. The results of this work can provide guiding significance for accurate calculation of wellbore pressure.

P13.01 Neuronal activity drives distinct invasion modes of glioma cells

Abstract BACKGROUND Gliomas are incurable brain tumors characterized by their infiltrative growth which makes them a whole-brain disease. Previously we described membrane protrusions called tumor microtubes (TMs), and glutamatergic synapses between neurons and glioma cells, as mechanisms contributing to glioma cell invasion and tumor progression. However, the interrelation of the two, and the exact mechanisms of glioma cell dynamics over time was unknown. Therefore, we investigate neuronal synaptic input on TM-associated glioma cell motility. MATERIAL AND METHODS Here we established a novel workflow for analyzing single glioma cell dynamics over several hours with in-vivo two-photon microscopy. First, a membranous fluorescent marking of patient-derived glioma cells was established to reliably track membrane changes. Secondly, augmented microscopy based on deep- and machine-learning algorithms was used to track glioma cells. Neuronal activity was manipulated with different doses of isoflurane anesthesia, and used to study its effects on glioma cell dynamics. RESULTS This novel method revealed that motility of glioma cells can be described by the displacement of whole glioma cell somata (somatokinesis) and TM dynamics. TM motility in turn could be sub-categorized into protrusion, retraction and branching. Next, we describe three different invasion modes, all with similarities to different cell types involved in CNS development. Lastly, the effects of neuronal activity on glioma cell invasion were investigated. With the application of high anesthesia and subsequently reduced neuronal activity, TM turnover, branching events and as a result glioma cell invasion were inhibited, but in a heterogeneous manner. CONCLUSION The novel workflow allowed to comprehensively characterize glioma cell invasion over several hours. Its application demonstrates novel, hitherto unknown cellular mechanisms of glioma cell invasion, and provides a link between TM biology and neuron-glioma communication. Finally, neuronal input drives distinct subtypes of glioma cell motility patterns.All in all, this work presents an important first step in understanding mechanisms that lead to the whole- brain colonization of glioma cells making these brain tumors incurable. A further characterization of the exact molecular mechanisms that drive neuronal activity-dependent glioma cell motility is warranted.

Research on Water Invasion Mode and Water Control Counter Measures of Geological MX Gas Reservoir

The MX gas reservoir is a typical carbonate gas reservoir and there are three types of water production characteristics in producing wells during the development process: fast rising type, slow rising type and stable type. The overall water invasion mode is dominated by the edge water invasion mode, with four types of water invasion modes including bottom water cone type, edge water finger type, edge water tongue type and native movable water tongue type. In this paper, numerical simulation techniques are used to investigate water management countermeasures for MX gas reservoirs based on the dynamic identification of water invasion. The results show that, after optimisation of the drainage and water control plans, the gas reservoir can produce steadily until the end of 2025 at a recovery rate of 3.06%, extending the time of steady production by about 9 months. In view of the relatively uniform advancement of fracture water channel and edge water into the interior of the gas reservoir, it is feasible to adopt active drainage and edge water control measures respectively to reduce the risk of water intrusion in the gas reservoir and ensure stable gas production.

Material balance method and classification of non-uniform water invasion mode for water-bearing gas reservoirs considering the effect of water sealed gas

In order to clarify the characteristics of non-uniform water invasion in water-bearing gas reservoirs, it is necessary to introduce the non-uniformity coefficient (A) and water invasion constant (B) to characterize the non-uniformity degree of reservoir physical properties and the activity degree of peripheral water, respectively, based on the dual mechanism of water invasion to recharge the formation energy and seal off the gas in the reservoir. Then, the material balance method considering the phenomenon of water sealed gas was established. On this basis, the water invasion characteristic curve chart of water-bearing gas reservoirs was plotted, and the non-uniform water invasion mode was classified based on the example gas reservoir. And the following research results were obtained. First, in the water invasion characteristic curve chart of water-bearing gas reservoirs which is plotted based on the material balance method considering the influence of water sealed gas, the upper right area and the lower left area are defined as recharge area and seal area, respectively. By taking A = 0 and B = 2 as the boundary, the recharge area is divided into strong recharge area and weak recharge area. By taking A = 2 and B = 2 as the boundary, the seal area is divided into strong seal area and weak seal area. And correspondingly there are four water invasion modes, i.e., strong recharge, weak recharge, weak seal and strong seal. Second, for fractured gas reservoirs, the non-uniformity degree of reservoir physical properties is high, and water sealed gas can be formed easily after water invasion. The dimensionless relative pseudo-pressure data of this type of gas reservoir is located in the seal area of the water invasion characteristic curve chart. Third, for the gas reservoirs whose reservoir physical properties are relatively uniform, the dimensionless relative pseudo-pressure data is located in the recharge area of the water invasion characteristic curve chart, and the recharge effect of water invasion on formation energy is greater than the weakening effect of water sealed gas on formation energy. Fourth, with the increase of A, the non-uniformity degree of reservoir physical properties increases, the water invasion characteristic curve shifts from the upper right to the lower left, and the recovery factor of gas reservoir decreases continuously. With the increase of B, the recharge effect of water invasion on formation energy and the weakening effect of water sealed gas on formation energy are both weakened, the distribution range of water invasion characteristic curve narrows to the recharge/seal boundary, and the corresponding range of gas reservoir recovery factor also narrows.

A preclinical pipeline to evaluate migrastatics as therapeutic agents in metastatic melanoma

BackgroundMetastasis is a hallmark of cancer and responsible for most cancer deaths. Migrastatics were defined as drugs interfering with all modes of cancer cell invasion and thus cancers’ ability to metastasise. First anti-metastatic treatments have recently been approved.MethodsWe used bioinformatic analyses of publicly available melanoma databases. Experimentally, we performed in vitro target validation (including 2.5D cell morphology analysis and mass spectrometric analysis of RhoA binding partners), developed a new traceable spontaneously metastasising murine melanoma model for in vivo validation, and employed histology (haematoxylin/eosin and phospho-myosin II staining) to confirm drug action in harvested tumour tissues.ResultsUnbiased and targeted bioinformatic analyses identified the Rho kinase (ROCK)-myosin II pathway and its various components as potentially relevant targets in melanoma. In vitro validation demonstrated redundancy of several RhoGEFs upstream of RhoA and confirmed ROCK as a druggable target downstream of RhoA. The anti-metastatic effects of two ROCK inhibitors were demonstrated through in vivo melanoma metastasis tracking and inhibitor effects also confirmed ex vivo by digital pathology.ConclusionsWe proposed a migrastatic drug development pipeline. As part of the pipeline, we provide a new traceable spontaneous melanoma metastasis model for in vivo quantification of metastasis and anti-metastatic effects by non-invasive imaging.