DMT1-dependent endosome-mitochondria interactions regulate mitochondrial iron translocation and metastatic outgrowth

Oncogene, Journal Year: 2024, Volume and Issue: 43(9), P. 650 - 667

Published: Jan. 6, 2024

Transient early endosome (EE)-mitochondria interactions can mediate mitochondrial iron translocation, but the associated mechanisms are still elusive. We showed that Divalent Metal Transporter 1 (DMT1) sustains translocation via EE-mitochondria in triple-negative MDA-MB-231, not luminal A T47D breast cancer cells. DMT1 silencing increases labile pool (LIP) levels and activates PINK1/Parkin-dependent mitophagy MDA-MB-231 Mitochondrial bioenergetics iron-associated protein profile were altered by rescued re-expression. Transcriptomic profiles upon strikingly different between 2D 3D culture conditions, suggesting environment context is crucial for knockout phenotype observed Lastly, vivo lung metastasis assay revealed promoted outgrowth of metastatic nodules both human murine models These findings reveal a DMT1-dependent pathway connecting to fitness

Language: Английский

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Spatiotemporal analysis of ratiometric biosensors in live multicellular spheroids using SPoRTS

Cell Reports Methods, Journal Year: 2025, Volume and Issue: unknown, P. 100987 - 100987

Published: Feb. 1, 2025

Language: Английский

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Inhibition of GPR68 induces ferroptosis and radiosensitivity in diverse cancer cell types

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: Feb. 3, 2025

Radioresistance is thought to be a major consequence of tumor milieu acidification resulting from the Warburg effect. Previously, using ogremorphin (OGM), small molecule inhibitor GPR68, an extracellular proton sensing receptor, we demonstrated that GPR68 key pro-survival pathway in glioblastoma cells. Here, demonstrate inhibition also induces ferroptosis lung cell carcinoma (A549) and pancreatic ductal adenocarcinoma (Panc02) Moreover, OGM synergized with ionizing radiation induce lipid peroxidation, hallmark ferroptosis, as well reduce colony size 2D 3D culture. not acutely detrimental but increases intracellular free ferrous iron, which known trigger reactive oxygen species (ROS) generation. In summary, peroxidation cancer cells sensitizes them part through mobilization iron. Our results suggest mediator radioresistance activated by acidic microenvironment.

Language: Английский

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Poly(2-alkyl-2-oxazoline) Hydrogels as Synthetic Matrices for Multicellular Spheroid and Intestinal Organoid Cultures

Biomacromolecules, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 3, 2025

The extracellular matrix (ECM) plays a crucial role in organoid cultures by supporting cell proliferation and differentiation. A key feature of the ECM is its mechanical influence on surrounding cells, directly affecting their behavior. Matrigel, most commonly used ECM, limited animal-derived origin, batch variability, uncontrollable properties, restricting use 3D cell-model-based mechanobiological studies. Poly(2-alkyl-2-oxazoline) (PAOx) synthetic hydrogels represent an appealing alternative because reproducibility versatile chemistry, enabling tuning hydrogel stiffness functionalization. Here, we studied PAOx with differing compressive moduli for potential to support growth. spheroid growth over several days without addition components. Furthermore, discovered intestinal epithelial polarity reversion demonstrate how tunable properties can be study effects morphology oxygenation live multicellular spheroids.

Language: Английский

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Fluorescent Molecular Probe for Imaging Hypoxia in 2D Cell Culture Monolayers and 3D Tumor Spheroids: The Cell Membrane Partition Model for Predicting Probe Distribution in a Spheroid

ACS Applied Materials & Interfaces, Journal Year: 2025, Volume and Issue: unknown

Published: March 13, 2025

Compared to cultured 2D cell monolayers, 3D multicellular spheroids are more realistic tumor models. Nonetheless, remain under-utilized in preclinical research, part, because there is a lack of fluorescence sensors that can noninvasively interrogate all the individual cells within spheroid. This present study describes deep-red fluorogenic molecular probe for microscopic imaging contain high level nitroreductase enzyme activity as biomarker hypoxia. A first-generation version produced "turn-on" monolayer under hypoxic conditions; however, it was not useful spheroid only accumulated peripheral cells. To guide structural optimization process, an intuitive theoretical membrane partition model conceived predict how dosed will distribute The identifies three limiting diffusion pathways determined by probe's properties. lipophilic with affinity rapidly becomes trapped membranes In contrast, very hydrophilic molecule negligible diffuses through intercellular space and rarely enters However, intermediate undergoes sequential out distributes Using predictive tool, second-generation fluorescent prepared smaller structure, optical sectioning using structured illumination or light sheet microscopy revealed roughly even throughout permeation likely be broadly applicable various classes molecules nanoparticles enable distribution

Language: Английский

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Protocol for in vivo fluorescence lifetime microendoscopic imaging of the murine femoral marrow

Methods in microscopy, Journal Year: 2025, Volume and Issue: unknown

Published: March 19, 2025

Abstract We present a protocol for micro-endoscopic fluorescence lifetime imaging in the femoral marrow of mice allowing analysis NAD(P)H-dependent metabolism at single cell level, vivo . Therefore, we employed gradient refractive index (GRIN) lens system fixed to mouse femur by specialized implant. provide step-by-step instructions practical usage microendoscopic implant and discuss experimental parameters required reliable NAD(P)H FLIM analysis, particularly referring photon statistics signal-to-noise ratio. Representative results indicate metabolic heterogeneity both tissue environment among LysM + myeloid cells expect here presented approach enable cellular functions dysfunctions, beyond metabolism, providing better understanding bone biology, health disease.

Language: Английский

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Black Phosphorus Nanosheets Incorporated Mesenchymal Stem Cell Spheroids for Accelerated Bone Regeneration

ACS Applied Nano Materials, Journal Year: 2025, Volume and Issue: unknown

Published: April 23, 2025

Language: Английский

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Oxygen-Sensitive Optical Nanosensors: Current Advances and Future Perspectives

ACS Sensors, Journal Year: 2025, Volume and Issue: unknown

Published: April 24, 2025

Oxygen sensing is essential across a wide range of fields, from understanding cellular metabolism and disease mechanisms to optimizing industrial environmental processes. In this Perspective, we highlight key developments in optical architectures (at the nanometer sub-micrometer scale), including their transduction methods applications vitro, vivo/in situ, nonbiological systems. We also discuss future directions for field domain expanding extra/intracellular sensing. address improving accessibility nonexpert users through need standardized protocols scalable production methods. Furthermore, advocate fostering interdisciplinary collaborations academic incubators, conference networking, strategic citation practices bridge gaps between fundamental research applied science expand impact these tools researchers outside field. Addressing challenges will help drive development more versatile widely accessible oxygen sensors, thus advancing innovation diverse disciplines.

Language: Английский

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Scalability of spheroid-derived small extracellular vesicles production in stirred systems

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: April 29, 2025

Small extracellular vesicle (sEV)-based therapies have gained widespread interest, but challenges persist to ensure standardization and high-scale production. Implementing upstream processes in a chemically defined media stirred-tank bioreactors (STBr) is mandatory closely control the cell environment, scale-up production, it remains significant challenge for anchorage-dependent cells. We used human β line, grown as monolayer or suspension spheroid stirred systems. assessed consequences of culturing these cells 3D with, without fetal bovine serum medium, growth, viability metabolism. next explored how different strategies might influence formation spinner flask, with aim transfer process instrumented Ambr®250 STBr. Lastly, we analyzed characterized sEV production monolayer, flask Generation spheroids medium allowed culture highly viable Spheroid size depended on system's volumetric power input (P/V), maintaining this parameter constant during proved be optimal strategy standardizing process. However, transferring (SpF) STBr at P/V modified size, due important geometric differences impeller design. Compared reference process, yield decreased two-fold SpF, increased Additionally, lower expression CD63 tetraspanin was observed produced both systems, suggesting reduced release exosomes compared ectosomes. This study addresses main issues encountered rather conducive

Language: Английский

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Multimodal Phasor Approach to Study Breast Cancer Cell Invasion in a 3D Spheroid Model

Chemical & Biomedical Imaging, Journal Year: 2025, Volume and Issue: unknown

Published: May 15, 2025

Language: Английский

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