Addressing the Challenges of High‐Throughput Cancer Tissue Proteomics for Clinical Application: ProCan

PROTEOMICS ◽  
2019 ◽  
Vol 19 (21-22) ◽  
pp. 1900109 ◽  
Author(s):  
Brett Tully ◽  
Rosemary L. Balleine ◽  
Peter G. Hains ◽  
Qing Zhong ◽  
Roger R. Reddel ◽  
...  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Débora Ferreira ◽  
Joaquim Barbosa ◽  
Diana A. Sousa ◽  
Cátia Silva ◽  
Luís D. R. Melo ◽  
...  

AbstractTriple-negative breast cancer is the most aggressive subtype of invasive breast cancer with a poor prognosis and no approved targeted therapy. Hence, the identification of new and specific ligands is essential to develop novel targeted therapies. In this study, we aimed to identify new aptamers that bind to highly metastatic breast cancer MDA-MB-231 cells using the cell-SELEX technology aided by high throughput sequencing. After 8 cycles of selection, the aptamer pool was sequenced and the 25 most frequent sequences were aligned for homology within their variable core region, plotted according to their free energy and the key nucleotides possibly involved in the target binding site were analyzed. Two aptamer candidates, Apt1 and Apt2, binding specifically to the target cells with $$K_{d}$$ K d values of 44.3 ± 13.3 nM and 17.7 ± 2.7 nM, respectively, were further validated. The binding analysis clearly showed their specificity to MDA-MB-231 cells and suggested the targeting of cell surface receptors. Additionally, Apt2 revealed no toxicity in vitro and showed potential translational application due to its affinity to breast cancer tissue sections. Overall, the results suggest that Apt2 is a promising candidate to be used in triple-negative breast cancer treatment and/or diagnosis.


2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
MI Khot ◽  
M Levenstein ◽  
R Coppo ◽  
J Kondo ◽  
M Inoue ◽  
...  

Abstract Introduction Three-dimensional (3D) cell models have gained reputation as better representations of in vivo cancers as compared to monolayered cultures. Recently, patient tumour tissue-derived organoids have advanced the scope of complex in vitro models, by allowing patient-specific tumour cultures to be generated for developing new medicines and patient-tailored treatments. Integrating 3D cell and organoid culturing into microfluidics, can streamline traditional protocols and allow complex and precise high-throughput experiments to be performed with ease. Method Patient-derived colorectal cancer tissue-originated organoidal spheroids (CTOS) cultures were acquired from Kyoto University, Japan. CTOS were cultured in Matrigel and stem-cell media. CTOS were treated with 5-fluorouracil and cytotoxicity evaluated via fluorescent imaging and ATP assay. CTOS were embedded, sectioned and subjected to H&E staining and immunofluorescence for ABCG2 and Ki67 proteins. HT29 colorectal cancer spheroids were produced on microfluidic devices using cell suspensions and subjected to 5-fluorouracil treatment via fluid flow. Cytotoxicity was evaluated through fluorescent imaging and LDH assay. Result 5-fluorouracil dose-dependent reduction in cell viability was observed in CTOS cultures (p<0.01). Colorectal CTOS cultures retained the histology, tissue architecture and protein expression of the colonic epithelial structure. Uniform 3D HT29 spheroids were generated in the microfluidic devices. 5-fluorouracil treatment of spheroids and cytotoxic analysis was achieved conveniently through fluid flow. Conclusion Patient-derived CTOS are better complex models of in vivo cancers than 3D cell models and can improve the clinical translation of novel treatments. Microfluidics can streamline high-throughput screening and reduce the practical difficulties of conventional organoid and 3D cell culturing. Take-home message Organoids are the most advanced in vitro models of clinical cancers. Microfluidics can streamline and improve traditional laboratory experiments.


Nanoscale ◽  
2017 ◽  
Vol 9 (36) ◽  
pp. 13592-13599 ◽  
Author(s):  
Qian Zhang ◽  
Ting Yin ◽  
Rongrong Xu ◽  
Wenjun Gao ◽  
Hui Zhao ◽  
...  

A self-designed high-throughput system has been developed for large-scale immuno-magnetic cell sorting of different T cells.


2008 ◽  
Vol 7 (4) ◽  
pp. 1412-1418 ◽  
Author(s):  
Ida Pucci-Minafra ◽  
Patrizia Cancemi ◽  
Nadia Ninfa Albanese ◽  
Gianluca Di Cara ◽  
Maria Rita Marabeti ◽  
...  

2018 ◽  
Vol 110 (1) ◽  
pp. 345-355 ◽  
Author(s):  
Jumpei Kondo ◽  
Tomoya Ekawa ◽  
Hiroko Endo ◽  
Kanami Yamazaki ◽  
Norio Tanaka ◽  
...  

2014 ◽  
Vol 445 (3) ◽  
pp. 584-590 ◽  
Author(s):  
Saet-byul Lee ◽  
Junhoe Cha ◽  
Im-kyung Kim ◽  
Joo Chun Yoon ◽  
Hyo Joon Lee ◽  
...  

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