The need to understand cancer biology at a cellular level requires a comprehensive interrogation of mRNA and proteins at high throughput and preferably simultaneously. The overall goal of the proposed research is to expand preliminary experiments by the applicant to further develop, validate and optimize magnetic nanosensors (magnetic relaxation switches, MRS) capable of sensing different types of molecular targets. In preliminary experiments we have shown that when monodisperse magnetic nanoparticles self-assemble into oligomeric nanoassemblies, there is a corresponding decrease in the spin-spin relaxation times (T2) of surrounding water detectable by NMR techniques. This fundamental observation has allowed us to measure DNA-DNA, protein-protein, small molecule protein interactions as well as enzyme activities (Nat Biotech 2002;20, 816-820). The magnetic relaxation switch (MRS) technique is exceedingly sensitive, allowing the detection of target molecules at femtomole levels with bench-top detectors and attomole levels by high-resolution MR imaging. The assay is performed in solution and does not require isolation or purification of the sample. In the proposed research the applicant will pursue three specific aims using telomerase as a cancer related target: (1) design and optimize MRS probes that independently recognize telomerase-mRNA, -protein and enzyme activity; (2) develop and optimize magnetic resonance techniques for simultaneous screening of telomerase-mRNA, -protein and -activity; and (3) develop and optimize internalization protocols for telomerase MRS probes that would allow detection of a target inside intact cells. It is hypothesized that telomerase mRNA, telomerase protein and telomerase activity can be detected in cell lysates and tissue extracts without prior purification or amplification using the developed MRS technique. Major biotechnical and medical applications of the developed technique lie in 1) the development of techniques to simultaneously interrogate RNA and proteins, 2) the development of high-throughput solution phase arrays, and 3) the ability to image molecular interactions by magnetic resonance imaging. The proposed research will be conducted in a multidisciplinary environment and the integrated training and research program will enable the candidate to establish himself as an independent cancer researcher. ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
7K01CA101781-03
Application #
6927337
Study Section
Subcommittee G - Education (NCI)
Program Officer
Ojeifo, John O
Project Start
2003-08-18
Project End
2008-07-31
Budget Start
2005-08-08
Budget End
2006-07-31
Support Year
3
Fiscal Year
2005
Total Cost
$135,270
Indirect Cost
Name
University of Central Florida
Department
Type
Organized Research Units
DUNS #
150805653
City
Orlando
State
FL
Country
United States
Zip Code
32826
Asati, Atul; Kaittanis, Charalambos; Santra, Santimukul et al. (2011) pH-tunable oxidase-like activity of cerium oxide nanoparticles achieving sensitive fluorigenic detection of cancer biomarkers at neutral pH. Anal Chem 83:2547-53
Santra, Santimukul; Kaittanis, Charalambos; Perez, J Manuel (2010) Aliphatic hyperbranched polyester: a new building block in the construction of multifunctional nanoparticles and nanocomposites. Langmuir 26:5364-73
Asati, Atul; Santra, Santimukul; Kaittanis, Charalambos et al. (2010) Surface-charge-dependent cell localization and cytotoxicity of cerium oxide nanoparticles. ACS Nano 4:5321-31
Santra, Santimukul; Kaittanis, Charalambos; Grimm, Jan et al. (2009) Drug/dye-loaded, multifunctional iron oxide nanoparticles for combined targeted cancer therapy and dual optical/magnetic resonance imaging. Small 5:1862-8
Asati, Atul; Santra, Santimukul; Kaittanis, Charalambos et al. (2009) Oxidase-like activity of polymer-coated cerium oxide nanoparticles. Angew Chem Int Ed Engl 48:2308-12
Kaittanis, Charalambos; Santra, Santimukul; Perez, J Manuel (2009) Role of nanoparticle valency in the nondestructive magnetic-relaxation-mediated detection and magnetic isolation of cells in complex media. J Am Chem Soc 131:12780-91
Nath, Sudip; Kaittanis, Charalambos; Ramachandran, Vasanth et al. (2009) Synthesis, magnetic characterization and sensing applications of novel dextran-coated iron oxide nanorods. Chem Mater 21:1761-1767
Kaittanis, Charalambos; Nath, Sudip; Perez, J Manuel (2008) Rapid nanoparticle-mediated monitoring of bacterial metabolic activity and assessment of antimicrobial susceptibility in blood with magnetic relaxation. PLoS One 3:e3253
Perez, J Manuel; Grimm, Jan; Josephson, Lee et al. (2008) Integrated nanosensors to determine levels and functional activity of human telomerase. Neoplasia 10:1066-72
Nath, Sudip; Kaittanis, Charalambos; Tinkham, Alisa et al. (2008) Dextran-coated gold nanoparticles for the assessment of antimicrobial susceptibility. Anal Chem 80:1033-8

Showing the most recent 10 out of 13 publications