Label-free image-based detection of drug resistance with optofluidic time-stretch microscopy (Conference Presentation)

2017 ◽  
Author(s):  
Hirofumi Kobayashi ◽  
Cheng Lei ◽  
Ailin Mao ◽  
Yiyue Jiang ◽  
Baoshan Guo ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Label Free

scholarly journals In situ drug-receptor binding kinetics in single cells: a quantitative label-free study of anti-tumor drug resistance

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Wei Wang ◽  
Linliang Yin ◽  
Laura Gonzalez-Malerva ◽  
Shaopeng Wang ◽  
Xiaobo Yu ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Receptor Binding ◽  
Single Cells ◽  
Binding Kinetics ◽  
Label Free ◽  
Tumor Drug Resistance ◽  
Drug Receptor

scholarly journals Label-Free Recognition of Drug Resistance via Impedimetric Screening of Breast Cancer Cells

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e57423 ◽  
Author(s):  
Bilge Eker ◽  
Robert Meissner ◽  
Arnaud Bertsch ◽  
Kapil Mehta ◽  
Philippe Renaud
Keyword(s):  
Breast Cancer ◽  
Drug Resistance ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Label Free

Proteomic Characterization of Circulating Extracellular Vesicles Identifies Novel Serum Myeloma Associated Markers

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1814-1814
Author(s):  
David Ciarlariello ◽  
Sean Harshman ◽  
Alessandro Canella ◽  
Erinn Hade ◽  
Alberto Rocci ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Protein Content ◽  
Cell Lines ◽  
Extracellular Vesicles ◽  
Stromal Cells ◽  
Plasma Cells ◽  
Predictive Biomarker ◽  
Total Serum ◽  
Label Free ◽  
Β2 Microglobulin

Abstract Introduction: Multiple myeloma (MM) is a hematological malignancy of clonal plasma cells (PCs) in the bone marrow (BM). MM cells are dependent on the BM microenvironment (e.g. BM stromal cells, macrophages etc) and create a network with surrounding cells. These cells play a pivotal role in the regulation of MM cell survival and drug resistance bydirect interactions through adhesion molecules causing cell adhesion mediated drug resistance (CAM-DR) or soluble factors including supportive cytokines (e.g. IL-6, IL-8, and VEGF) or exosomes (or extracellular vesicles). Exosomes (EV) are endosome-derived membrane-covered cell fragments, which contain specific protein and RNA cargo. The protein content of EVs in MM has not been readily explored. Recently our group established the use a global label-free quantification method to determine the relative amount of proteins identified from EVs obtained from MM cell lines. Here we are reporting a systematic proteomic analysis of EVs derived from MM cell lines, blood from MM patients, and BM from MM patients. We also report preliminary data showing the biological importance of specific EV enriched proteins in MM cell lines and MM patients. Methods: Liquid Chromatography Mass Spectrometry (MS) and Label-free relative quantitation were used to assess the protein content of MM EV and cellular lysates. Cryo-Transmission Electron Microscopy (cryo-TEM) and Nanoparticle Tracking Analysis were used to assess size distribution and specific EV surface markers in EV isolated from the supernatant of MM cell lines and from the serum of non-cancer, MGUS, smoldering MM and active MM donors. Western blot and enzyme-linked immunosorbent assays were used to validate MS data in a bigger cohort of primary MM patients and to assess the biological effect of MM derived EV in the BM stromal cells. Results: Our data show that the Major Histocompatibility Complex Class I (MHCI) and its associated binding protein β2-microglobulin (β2-MG) are the most abundant communally enriched proteins in the EV derived from MM cell lines and from the serum of MM patients. Although it is well known that the serum level of β2-MG is an important prognostic factor in MM, our data indicate that β2-MG represents only a small percentage of the total serum β2-MG, suggesting that two separate β2-microglobulin populations coexist in the serum of patients. Additionally, we show that the main receptor of hyaluronic acid, CD44, is highly expressed in the EVs isolated from the corticosteroid resistant MM cell line, MM.1R, and is a protein that is differentially expressed in EVs isolated from newly diagnosed MM patients and which seems to play a key role in communicating with the BM microenvironment. By using a large cohort of serum obtained from MM patients prospectively treated on a randomized phase 3 trial (233), we establish the potential of serum CD44 as a predictive biomarker of overall survival. These results support the analysis of EVs as easily accessible MM biomarker. Conclusions: Our results generate a foundation for the potential use of circulating EVs as novel serum markers of MM and provide the rationale to further explore previously unconsidered molecular players associated with MM disease. Disclosures Palumbo: Array BioPharma: Consultancy; Onyx Pharmaceuticals: Consultancy; Millennium Pharmaceuticals Inc., a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Consultancy, Honoraria; Janssen-Cilag: Consultancy, Honoraria; Genmab A/S: Consultancy; Bristol-Myers Squibb: Consultancy; Amgen: Consultancy; Sanofi Aventis: Consultancy.

A SYSTEMS BIOLOGY APPROACH TO THE STUDY OF CISPLATIN DRUG RESISTANCE IN OVARIAN CANCERS

2007 ◽  
Vol 05 (02a) ◽  
pp. 383-405 ◽  
Author(s):  
JAKE Y. CHEN ◽  
ZHONG YAN ◽  
CHANGYU SHEN ◽  
DAWN P. G. FITZPATRICK ◽  
MU WANG
Keyword(s):  
Drug Resistance ◽  
Systems Biology ◽  
Protein Interactions ◽  
Computational Study ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Differential Proteomics ◽  
Acquired Drug Resistance ◽  
Significant Enrichment ◽  
Initial List

Cisplatin-induced drug resistance is known to involve a complex set of cellular changes whose molecular mechanism details remain unclear. In this study, we developed a systems biology approach to examine proteomics- and network-level changes between cisplatin-resistant and cisplatin-sensitive cell lines. This approach involves experimental investigation of differential proteomics profiles and computational study of activated enriched proteins, protein interactions, and protein interaction networks. Our experimental platform is based on a Label-free liquid Chromatography/mass spectrometry proteomics platform. Our computational methods start with an initial list of 119 differentially expressed proteins. We expanded these proteins into a cisplatin-resistant activated sub-network using a database of human protein-protein interactions. An examination of network topology features revealed the activated responses in the network are closely coupled. By examining sub-network proteins using gene ontology categories, we found significant enrichment of proton-transporting ATPase and ATP synthase complexes activities in cisplatin-resistant cells in the form of cooperative down-regulations. Using two-dimensional visualization matrixes, we further found significant cascading of endogenous, abiotic, and stress-related signals. Using a visual representation of activated protein categorical sub-networks, we showed that molecular regulation of cell differentiation and development caused by responses to proteome-wide stress as a key signature to the acquired drug resistance.

Label-free, mass-sensitive single-molecule imaging using interferometric scattering microscopy

2020 ◽  
Author(s):  
Nikolas Hundt
Keyword(s):  
Single Molecule ◽  
Cellular Systems ◽  
Single Molecule Imaging ◽  
Label Free ◽  
Measurement Sensitivity ◽  
Mass Distributions ◽  
Quantitative Measurements ◽  
Contrast Mechanism ◽  
Cellular Components ◽  
Scattering Signal

Abstract Single-molecule imaging has mostly been restricted to the use of fluorescence labelling as a contrast mechanism due to its superior ability to visualise molecules of interest on top of an overwhelming background of other molecules. Recently, interferometric scattering (iSCAT) microscopy has demonstrated the detection and imaging of single biomolecules based on light scattering without the need for fluorescent labels. Significant improvements in measurement sensitivity combined with a dependence of scattering signal on object size have led to the development of mass photometry, a technique that measures the mass of individual molecules and thereby determines mass distributions of biomolecule samples in solution. The experimental simplicity of mass photometry makes it a powerful tool to analyse biomolecular equilibria quantitatively with low sample consumption within minutes. When used for label-free imaging of reconstituted or cellular systems, the strict size-dependence of the iSCAT signal enables quantitative measurements of processes at size scales reaching from single-molecule observations during complex assembly up to mesoscopic dynamics of cellular components and extracellular protrusions. In this review, I would like to introduce the principles of this emerging imaging technology and discuss examples that show how mass-sensitive iSCAT can be used as a strong complement to other routine techniques in biochemistry.

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