This is a competing renewal application that builds on the instrumentation and expertise developed in the course of the previous funding periods. We propose to investigate the feasibility of extending the current magnetic cell separation methods to applications relying on the cell's own, natural (intrinsic) magnetization. The application rests on the hypothesis that there is a link between malignant cell transformation and an increase in the cell magnetic susceptibility. The supporting evidence comes from a large body of data from other studies on the magnetic properties of tissues, in particular, from electron (spin) paramagnetic resonance, EPR, and studies of solid tumors. Additional supporting evidence comes from our own data on the magnetic field-induced motion (magnetophoresis) in physiologic electrolyte solutions of cancer cell lines (HeLa and Hep 3B), that demonstrate statistically greater velocity compared to controls (oxygenated red blood cells). We have shown an additional increase in the cell magnetophoretic mobility following the addition of soluble iron compounds to the culture media, suggesting an effect of intracellular iron uptake on the cell magnetization. The proposed effort is divided between:
Specific Aims : SA1: To investigate the molecular mechanism for the observed increase in magnetophoretic mobility (MM) in selected cancer cell lines as compared to matched normal cells. In collaboration with experts in cell tracking velocimetry (Dr. Chalmers) and EPR spectrometry (Dr. Kuppusamy) at the subcontract site (The Ohio State University), the effect of iron in the medium on the cell magnetophoretic mobility and the increased intracellular paramagnetic content will be investigated on cancer cell lines. SA2: To improve the sensitivity and resolution of the cell magnetophoretic mobility analyzer, the Cell Tracking Velocimetry (CTV). The increase of magnetophoretic mobility resolution by a factor up to 10 fold will be accomplished by increasing the magnetic energy density and field gradient of the apparatus, by improving the imaging capability of the cell tracking hardware and software, and by including corrections for fluid dynamics artifacts in collaboration with an expert in field-flow fractionation (Dr. Williams). SA3: To measure the magnetic susceptibility of selected primary tumors at a single cell level against a baseline of normal blood cells. Primary tumor cell lines will be tested against normal controls for differences in the magnetophoretic mobility, in collaboration with an expert oncologist (Dr. Borden). SA4: To test the feasibility of the label-less, magnetic cancer cell separation on model blood cell suspensions. The proposed strategy is based on the soluble - rather than particulate - iron transport processes, and may therefore be more finely tuned to the pathobiology of the cancer cell than is practiced today. It could have a profound impact on how the magnetic cell separation is practiced and utilized and could complement and extend the applications of the already existing magnetic cell separation methods.

Public Health Relevance

There is an intense interest in identifying cancer biomarkers for improved diagnosis and prognosis in the treatment of the disease. One particularly promising application is the detection, enumeration and analysis of circulating tumor cells (CTC's) in blood. There is evidence of abnormal paramagnetic contributions to the magnetic susceptibility of cells that have undergone malignant transformation coming from the laboratories collaborating on this grant application. We propose to systematically investigate this effect by using our state-of-the-art magnetic research facilities that include the magnetic cell separation laboratory and electron (spin) paramagnetic resonance (EPR) spectrometry facility available at the primary (Cleveland Clinic) and the subcontract (The Ohio State University) sites. The proposed research could have a profound impact on the magnetic separation methods in application to biomedical research and the treatment of cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA062349-20
Application #
8443349
Study Section
Special Emphasis Panel (ZRG1-BST-Z (02))
Program Officer
Ossandon, Miguel
Project Start
1994-02-18
Project End
2016-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
20
Fiscal Year
2013
Total Cost
$336,161
Indirect Cost
$69,115
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Lustberg, Maryam B; Balasubramanian, Priya; Miller, Brandon et al. (2014) Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients. Breast Cancer Res 16:R23
Moore, Lee R; Williams, P Stephen; Nehl, Franziska et al. (2014) Feasibility study of red blood cell debulking by magnetic field-flow fractionation with step-programmed flow. Anal Bioanal Chem 406:1661-70
Joshi, Powrnima; Jacobs, Barbara; Derakhshan, Adeeb et al. (2014) Enrichment of circulating melanoma cells (CMCs) using negative selection from patients with metastatic melanoma. Oncotarget 5:2450-61
Orita, Toru; Moore, Lee R; Joshi, Powrnima et al. (2013) A quantitative determination of magnetic nanoparticle separation using on-off field operation of quadrupole magnetic field-flow fractionation (QMgFFF). Anal Sci 29:761-4
Wu, Yongqi; Deighan, Clayton J; Miller, Brandon L et al. (2013) Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology. Methods 64:169-82
Joshi, Powrnima; Zborowski, Maciej; Triozzi, Pierre L (2013) Circulating melanoma cells: scoping the target. Front Oncol 3:189
Jin, Xiaoxia; Chalmers, Jeffrey J; Zborowski, Maciej (2012) Iron transport in cancer cell culture suspensions measured by cell magnetophoresis. Anal Chem 84:4520-6
Jin, Xiaoxia; Yazer, Mark H; Chalmers, Jeffrey J et al. (2011) Quantification of changes in oxygen release from red blood cells as a function of age based on magnetic susceptibility measurements. Analyst 136:2996-3003
Zborowski, Maciej; Chalmers, Jeffrey J (2011) Rare cell separation and analysis by magnetic sorting. Anal Chem 83:8050-6
Hoyos, Mauricio; Moore, Lee; Williams, P Stephen et al. (2011) The use of a linear Halbach array combined with a step-SPLITT channel for continuous sorting of magnetic species. J Magn Magn Mater 323:1384-1388

Showing the most recent 10 out of 65 publications