title: NAD(P)H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes.
abstract: During the development of neoplasia, epithelial tissues undergo biochemical and structural changes that can manifest in tissue fluorescence. There have been several reports on different in vivo fluorescence characteristics between normal and precancerous (dysplastic) tissues. However, it has been difficult to identify and quantify the origins of these changes, mainly because of distortions introduced in measured tissue fluorescence spectra by tissue scattering and absorption. Such distortions can be removed by combining information in simultaneously measured fluorescence and reflectance spectra. Thus, we can recover the intrinsic (undistorted) tissue fluorescence. In this report, we show that extraction of the intrinsic fluorescence allows us: (a) to determine the fluorescence spectra of NAD(P)H and collagen in an in vivo environment, and (b) to use these NAD(P)H and collagen spectra to describe, quantitatively, diagnostically significant biochemical changes between normal and dysplastic tissues. Specifically, by analyzing intrinsic fluorescence of human epithelial tissue as it becomes deoxygenated in vivo, we can resolve the fluorescence spectra of NAD(P)H and collagen, two of the major tissue fluorophores. This is important because fluorescence depends on the local environment of the chromophore. Then, we extract the intrinsic fluorescence spectra of sites from 35 patients with suspected cervical lesions and 7 patients with Barrett's esophagus and describe them accurately as a linear combination of NAD(P)H and collagen contributions. In both tissue cases, we find that low collagen and high NAD(P)H fluorescence characterizes the high-grade dysplastic lesions when compared with nondysplastic tissues. These data present evidence for the presence of detectable levels of NAD(P)H fluorescence in human epithelial tissues in an in vivo setting and demonstrate that NAD(P)H and collagen may be used as quantitative fluorescence biomarkers for in vivo detection of dysplasia in the cervix and the esophagus.
authors.affiliation: G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Irene
authors.initials: I
authors.lastname: Georgakoudi
authors.forename: Brian C
authors.initials: BC
authors.lastname: Jacobson
authors.forename: Markus G
authors.initials: MG
authors.lastname: Müller
authors.forename: Ellen E
authors.initials: EE
authors.lastname: Sheets
authors.forename: Kamran
authors.initials: K
authors.lastname: Badizadegan
authors.forename: David L
authors.initials: DL
authors.lastname: Carr-Locke
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Crum
authors.forename: Charles W
authors.initials: CW
authors.lastname: Boone
authors.forename: Ramachandra R
authors.initials: RR
authors.lastname: Dasari
authors.forename: Jacques
authors.initials: J
authors.lastname: Van Dam
authors.forename: Michael S
authors.initials: MS
authors.lastname: Feld
mesh_terms: D001471:Barrett Esophagus / Q000378:metabolism / Q000473:pathology; D014408:Biomarkers, Tumor / Q000378:metabolism*; D015687:Cell Hypoxia / Q000502:physiology; D002584:Cervix Uteri / Q000378:metabolism / Q000473:pathology; D003094:Collagen / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology; D004938:Esophageal Neoplasms / Q000378:metabolism* / Q000473:pathology; D004947:Esophagus / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D009249:NADP / Q000378:metabolism*; D011230:Precancerous Conditions / Q000378:metabolism* / Q000473:pathology; D013050:Spectrometry, Fluorescence; D002578:Uterine Cervical Dysplasia / Q000378:metabolism / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D009249:NADP; D003094:Collagen