Biodistribution of cisplatin revealed by imaging mass cytometry identifies extensive collagen binding in tumor and normal tissues

Chang, Qing; Ornatsky, Olga; Siddiqui, Iram; Straus, Rita; Baranov, Vladimir; Hedley, David W.

doi:10.1038/srep36641

Cited by 90 publications

(100 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With the use of metal tags instead of fluorophores, MISH overcomes several significant hurdles of traditional protocols and will be amenable to high-dimensional studies. Moreover, with the recent developments of imaging mass cytometry on tissue sections (25), MISH has the potential of being applicable to a variety of sample types.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, with the recent developments of imaging mass cytometry on tissue sections (25), MISH has the potential of being applicable to a variety of sample types. This function can be accomplished through synthesis of RNA and through control of its stability and location.…”

Section: Technical Notementioning

confidence: 99%

“…Imaging mass cytometry data acquisition and visualization were performed according to a procedure described in (25). Briefly, stained and air-dried slides were inserted into the ablation chamber of the Hyperion TM Imaging System (Fluidigm), where a pulsed 200 Hz laser is focused to a 1 mm spot and applied over a user-defined area, ablating adjacent spots in 1 mm steps as the slide moves under the laser beam.…”

Section: Mass Cytometry Data Acquisition and Analysismentioning

confidence: 99%

“…The plumes of vaporized material are streamed by inert gas with high time fidelity into the inductively coupled plasma ion source for analysis by the mass cytometer. Data processing and visualization were performed using in-house-developed Wolfram Mathematica V R (V10.3) algorithms (25). Images of each mass channel were reconstructed by plotting the laser shot signals in the order in which they were recorded, line scan by line scan.…”

Section: Mass Cytometry Data Acquisition and Analysismentioning

confidence: 99%

“…This technique overcomes the issue of spectral overlap that limits the use of fluorescent labels and provides researchers with the ability to measure >40 markers in a single cell. Mass cytometry is gaining acceptance in various research fields such as immunology (21,22), oncology (23)(24)(25)(26), and in clinical analyses (27,28).…”

mentioning

confidence: 99%

See 4 more Smart Citations

Simultaneous Detection of Protein and mRNA in Jurkat and KG‐1a Cells by Mass Cytometry

Mavropoulos

Allo

et al. 2017

Cytometry Pt A

View full text Add to dashboard Cite

Mass cytometry uniquely enables high-dimensional single-cell analysis of complex populations. This recently developed technology is based on inductively coupled time-of-flight mass spectrometry for multiplex proteomic analysis of more than 40 markers per cell. The ability to characterize the transcriptome is critical for the understanding of disease pathophysiology, medical diagnostics, and drug discovery. Current techniques allowing the in situ detection of transcripts in single cells are limited to a small number of simultaneous targets and are generally tedious and labor-intensive. In this report, we present the development of a multiplex method for targeted RNA detection by combining the mass cytometry and RNAscope® platforms. This novel assay, called Metal In Situ Hybridization (MISH), includes the hybridization of RNA-specific target probes followed by signal amplification achieved through a cascade of hybridization events, ending with the binding of amplifier-specific detector probes. The detector probes are tagged with isotopically pure metal atoms used for detection by mass cytometry. Proof-of-principle experiments show the simultaneous detection of three mRNA targets in Jurkat cells in suspension cell assay mode. The localization of transcripts was also investigated using the imaging mass cytometry platform in Jurkat and KG-1a cells. In addition, we optimized the antibody staining procedure to allow the co-detection of mRNA and cell surface markers. Our data demonstrate that MISH can be used to complement protein detection by mass cytometry as well as to investigate gene transcription and translation in single cells. © 2017 International Society for Advancement of Cytometry.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Technical Notementioning

confidence: 99%

Section: Mass Cytometry Data Acquisition and Analysismentioning

confidence: 99%

Section: Mass Cytometry Data Acquisition and Analysismentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Simultaneous Detection of Protein and mRNA in Jurkat and KG‐1a Cells by Mass Cytometry

Mavropoulos

Allo

et al. 2017

Cytometry Pt A

View full text Add to dashboard Cite

show abstract

Profiling the 3D interaction between germ cell tumors and microenvironmental cells at the transcriptome and secretome level

et al. 2022

View full text Add to dashboard Cite

The tumor microenvironment (TM), consisting of the extracellular matrix (ECM), fibroblasts, endothelial cells, and immune cells, might affect tumor invasiveness and the outcome of standard chemotherapy. This study investigated the cross talk between germ cell tumors (GCT) and surrounding TM cells (macrophages, T‐lymphocytes, endothelial cells, and fibroblasts) at the transcriptome and secretome level. Using high‐throughput approaches of three‐dimensional (3D) co‐cultured cellular aggregates, this study offers newly identified pathways to be studied with regard to sensitivity toward cisplatin‐based chemotherapy or tumor invasiveness as a consequence of the cross talk between tumor cells and TM components. Mass‐spectrometry‐based secretome analyses revealed that TM cells secreted factors involved in ECM organization, cell adhesion, angiogenesis, and regulation of insulin‐like growth factor (IGF) transport. To evaluate direct cell–cell contacts, green fluorescent protein (GFP)‐expressing GCT cells and mCherry‐expressing TM cells were co‐cultured in 3D. Afterward, cell populations were separated by flow cytometry and analyzed by RNA sequencing. Correlating the secretome with transcriptome data indicated molecular processes such as cell adhesion and components of the ECM being enriched in most cell populations. Re‐analyses of secretome data with regard to lysine‐ and proline‐hydroxylated peptides revealed a gain in proteins, such as collagens and fibronectin. Cultivation of GCT cells on collagen I/IV‐ or fibronectin‐coated plates significantly elevated adhesive and migratory capacity, while decreasing cisplatin sensitivity of GCT cells. Correspondingly, cisplatin sensitivity was significantly reduced in GCT cells under the influence of conditioned medium from fibroblasts and endothelial cells. This study sheds light on the cross talk between GCT cells and their circumjacent TM, which results in deposition of the ECM and eventually promotes a pro‐tumorigenic environment through enhanced migratory and adhesive capacity, as well as decreased cisplatin sensitivity. Hence, our observations indicate that targeting the ECM and its cellular components might be a novel therapeutic option in combination with cisplatin‐based chemotherapy for GCT patients.

show abstract

Multidimensional profiling of drug‐treated cells by Imaging Mass Cytometry

2019

View full text Add to dashboard Cite

In pharmaceutical research, high‐content screening is an integral part of lead candidate development. Measuring drug response in vitro by examining over 40 parameters, including biomarkers, signaling molecules, cell morphological changes, proliferation indices, and toxicity in a single sample, could significantly enhance discovery of new therapeutics. As a proof of concept, we present here a workflow for multidimensional Imaging Mass Cytometry™ (IMC™) and data processing with open source computational tools. CellProfiler was used to identify single cells through establishing cellular boundaries, followed by histoCAT™ (histology topography cytometry analysis toolbox) for extracting single‐cell quantitative information visualized as t‐SNE plots and heatmaps. Human breast cancer‐derived cell lines SKBR3, HCC1143, and MCF‐7 were screened for expression of cellular markers to generate digital images with a resolution comparable to conventional fluorescence microscopy. Predicted pharmacodynamic effects were measured in MCF‐7 cells dosed with three target‐specific compounds: growth stimulatory EGF, microtubule depolymerization agent nocodazole, and genotoxic chemotherapeutic drug etoposide. We show strong pairwise correlation between nuclear markers pHistone3 S28 , Ki‐67, and p4E‐BP1 T37/T46 in classified mitotic cells and anticorrelation with cell surface markers. Our study demonstrates that IMC data expand the number of measured parameters in single cells and brings higher‐dimension analysis to the field of cell‐based screening in early lead compound discovery.

show abstract

Biodistribution of cisplatin revealed by imaging mass cytometry identifies extensive collagen binding in tumor and normal tissues

Cited by 90 publications

References 29 publications

Simultaneous Detection of Protein and mRNA in Jurkat and KG‐1a Cells by Mass Cytometry

Simultaneous Detection of Protein and mRNA in Jurkat and KG‐1a Cells by Mass Cytometry

Profiling the 3D interaction between germ cell tumors and microenvironmental cells at the transcriptome and secretome level

Multidimensional profiling of drug‐treated cells by Imaging Mass Cytometry

Contact Info

Product

Resources

About