While altered cell metabolism is an emerging hallmark of cancer, there is a crucial need of new tools for quantitation of metabolites. Though NMR spectroscopy, mass spectrometry, and Raman spectroscopy are widely used for molecular detection in tissue extracts or intact tissues, these tools do not tell the spatial locations of the analytes inside the cell. We address this unmet need via development of multiplex stimulated Raman scattering (SRS) microscopy to enable quantitative vibrational imaging of metabolites in live tumor cells and intact biopsies. The recently developed SRS microscopy allows high-speed, high-sensitivity imaging of single Raman bands in live cells. However, the single-frequency SRS imaging technique has limited capability because it cannot resolve molecular species that often have overlapped Raman bands. We propose to overcome this technical barrier via parallel detection of spectrally dispersed SRS signals enabled by a homebuilt tuned amplifier array. In a pilot study, we demonstrated multiplex SRS imaging of live pancreatic cancer cells with a pixel dwell time of 40 ?s.
In Aim 1, we will develop multiplex stimulated Raman loss (SRL) microscopy and multivariate analysis algorithm to enable quantitative vibrational imaging of lipid metabolites in live cells.
In Aim 2, we will develop epi-detected multiplex SRL microscopy to enable high-speed, large-area spectroscopic imaging of tumor biopsies. By accomplishment of the two aims, we will generate a high- sensitivity, high-speed, spectral imaging platform for molecular analysis of live cells with sub-micron spatial resolution. This platform will permit label-free visualization of metabolic conversion in live cancr cells, which is not possible with proteomics tools. Such capability is critical for mechanistic understanding of cancer metabolism and precise evaluation of drugs targeting cancer metabolism. This platform will also permit large- area mapping of intact tumor biopsies and offer information about metabolic biomarkers (e.g. cholesteryl ester) that are indicative of cancer aggressiveness.

Public Health Relevance

Aberrant expressions of lipogenic genes have been found in brain, mammary, prostate and many other cancers. Nonetheless, lipid metabolism has not been used as a prognostic factor for cancer aggressiveness due to lack of differential detection and quantitation technology. Our spectroscopic imaging technology is able to fill this gap by quantifying the amount and composition of intracellular lipid droplets, an important aspect of lipogenesis in tumor cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA182608-02
Application #
8738630
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Knowlton, John R
Project Start
2013-09-20
Project End
2016-08-31
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Purdue University
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
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Yue, Shuhua; Cheng, Ji-Xin (2016) Deciphering single cell metabolism by coherent Raman scattering microscopy. Curr Opin Chem Biol 33:46-57
Liao, Chien-Sheng; Slipchenko, Mikhail N; Wang, Ping et al. (2015) Microsecond Scale Vibrational Spectroscopic Imaging by Multiplex Stimulated Raman Scattering Microscopy. Light Sci Appl 4:
Zhang, Chi; Zhang, Delong; Cheng, Ji-Xin (2015) Coherent Raman Scattering Microscopy in Biology and Medicine. Annu Rev Biomed Eng 17:415-45
Lee, Hyeon Jeong; Zhang, Wandi; Zhang, Delong et al. (2015) Assessing cholesterol storage in live cells and C. elegans by stimulated Raman scattering imaging of phenyl-Diyne cholesterol. Sci Rep 5:7930
Liao, Chien-Sheng; Choi, Joon Hee; Zhang, Delong et al. (2015) Denoising Stimulated Raman Spectroscopic Images by Total Variation Minimization. J Phys Chem C Nanomater Interfaces 119:19397-19403
Yue, Shuhua; Li, Junjie; Lee, Seung-Young et al. (2014) Cholesteryl ester accumulation induced by PTEN loss and PI3K/AKT activation underlies human prostate cancer aggressiveness. Cell Metab 19:393-406
Zhang, Delong; Wang, Ping; Slipchenko, Mikhail N et al. (2014) Fast vibrational imaging of single cells and tissues by stimulated Raman scattering microscopy. Acc Chem Res 47:2282-90