We successfully brought nanoFACS, a method for analyzing and sorting naonoscale particles, to NIH, by modifying the Influx flow cytometer in a NCI/CCR core FACS facility in Building 10. This now allows us to analyze and sort exosomes and microparticles as small as approximately 100nm. We have developed and tested numerous modifications of the technique and instrument configuration to improve the detection limits to routinely analyze and sort particles as small as 40nm (which would span the full range of biologically significant exosome sizes). We also have obtained a new instrument, a Beckman Coutler MoFlo AstriosEQ,with modifications to further improve our analystical and sorting capabilities, as part of a CRADA established between our group and Beckman. Over the next year, we will continue 1) to use this technology to identify novel biomarkers that we can use to guide and monitor the effects of cancer therapies and immune responses (to vaccines or other immunotherapies), and 2) to identify subsets of these nanoparticles and determine their functions in tumor biology, immunobiology, and other aspects of biology.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011502-02
Application #
8938164
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Wiklander, Oscar P B; Bostancioglu, R Beklem; Welsh, Joshua A et al. (2018) Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures. Front Immunol 9:1326
Musich, Thomas; Jones, Jennifer C; Keele, Brandon F et al. (2017) Flow virometric sorting and analysis of HIV quasispecies from plasma. JCI Insight 2:e90626
Nolan, John P; Jones, Jennifer C (2017) Detection of platelet vesicles by flow cytometry. Platelets 28:256-262
Datta, Amrita; Kim, Hogyoung; Lal, Madhu et al. (2017) Manumycin A suppresses exosome biogenesis and secretion via targeted inhibition of Ras/Raf/ERK1/2 signaling and hnRNP H1 in castration-resistant prostate cancer cells. Cancer Lett 408:73-81
Morales-Kastresana, Aizea; Telford, Bill; Musich, Thomas A et al. (2017) Labeling Extracellular Vesicles for Nanoscale Flow Cytometry. Sci Rep 7:1878
Morales-Kastresana, Aizea; Jones, Jennifer C (2017) Flow Cytometric Analysis of Extracellular Vesicles. Methods Mol Biol 1545:215-225
Watson, Dionysios C; Bayik, Defne; Srivatsan, Avinash et al. (2016) Efficient production and enhanced tumor delivery of engineered extracellular vesicles. Biomaterials 105:195-205
Danielson, Kirsty M; Estanislau, Jessica; Tigges, John et al. (2016) Diurnal Variations of Circulating Extracellular Vesicles Measured by Nano Flow Cytometry. PLoS One 11:e0144678