Whole-body optical imaging of gene expression and uses thereof

#The invention relates to the whole-body external optical imaging of gene expression. Specifically, methods for whole-body external optical imaging of gene expression and methods for evaluating a candidate protocol or drug for treating diseases or disorders using a fluorophore operatively linked to the promoter of a gene and external optical imaging are provided herein. Methods to screen for substances or genes that regulate target promoters are also provided.

#Whole-body imaging technology has been used to monitor “tracer molecules” in the intact body. For example, Brenner et al. studied the diagnostic value of iodine-123-2-hydroxy-3-iodo-6-methoxy-N-[(1-ethyl-2-pyrrolidinyl)methyl] benzamide (IBZM) whole-body imaging in comparison to thallium-201 scintigraphy in patients with metastatic malignant melanoma (Brenner et al., Eur. J. Nucl. Med., 26(12):1567-71 (1999)). Benard et al. conducted clinical evaluation of processing techniques for attenuation correction with 137Cs in whole-body PET imaging (Benard et al., J. Nucl. Med., 40(8):1257-63 (1999)). Jerusalem et al. showed that whole-body positron emission tomography using 18F-fluorodeoxyglucose for posttreatment evaluation in Hodgkin's disease and non-Hodgkin's lymphoma has higher diagnostic and prognostic value than classical computed tomography scan imaging (Jerusalem et al., Blood, 94(2):429-33 (1999)). Eustace et al. discussed practical issues, clinical applications, and future directions of whole-body MR imaging (Eustace et al., Magn. Reson. Imaging Clin. (N. Am), 7(2):209-36 (1999)). Engelson et al. studied fat distribution in HIV-infected patients reporting truncal enlargement quantified by whole-body magnetic resonance imaging (Engelson et al., Am. J. Clin. Nutr., 69(6 :1162-9 (1999)). Valk et al. used whole-body positron emission tomography (PET) imaging with [18F]fluorodeoxyglucose in management of recurrent colorectal cancer (Valk et al., Arch. Surg., 134(5):503-11 (1999)). Saunders et al. evaluated fluorine-18-fluorodeoxyglucose whole body positron emission tomography imaging in the staging of lung cancer (Saunders et al., Ann. Thorac. Surg., 67(3):790-7 (1999)).

#U.S. Pat. No. 5,650,135 discloses a noninvasive method for detecting the localization of an entity under study from within a mammalian subject, which method comprises: (a) administering to the subject a conjugate of the entity and a light-generating moiety or a transformed cell expressing the light-generating moiety; (b) after a period of time in which the conjugate or transformed cell can achieve localization in the subject, immobilizing the subject within the detection field of a photodetector device; (c) maintaining the subject in an immobilized condition, (d) during said maintaining, measuring photon emission from the light-generating moiety, localized in the subject, with the photodetector device until an image of photon emission can be constructed; and (e) detecting said image through an opaque tissue of said mammal. U.S. Pat. No. 5,650,135 also discloses a noninvasive method for detecting the level of an entity under study in a mammalian subject over time, which method comprises: (a) administering to the subject a conjugate of the entity and a light-generating moiety or a transformed cell expressing the light-generating moiety; (b) placing the subject within the detection field of a photodetector device; (c) maintaining the subject in the detection field of the device; (d) during said maintaining, measuring photon emission from the light-generating moiety, in the subject, with the photodetector device; and (e) repeating steps (b) through (d) at selected intervals, wherein said repeating is effective to detect changes in the level of the entity in the subject over time.

#FIGS. 1A and 1B show the time course of expression of adenoviral-administered GFP in brain and liver respectively. Fluorescence first becomes visible in the brain within six (6) hours after local delivery and liver fluorescence became detectable at about seven (7) hours after injection into the tail vein.

#FIGS. 2A and 2B art pertinent to administration of lentiviral vectors. FIG. 2A is a diagram of lentiviral vector GFP-LV. FIG. 2B is a diagram of a control observation method; whole body measurement involved use of a light box.

#A. Definitions

#Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this invention belongs. All patents, applications, published applications and other publications and sequences from GenBank and other data bases referred to herein are incorporated by reference in their entirety.

#As used herein, “delivering a nucleic acid to a multi-cellular organism” refers to a process in which the nucleic acid is either administered directly into the body of the multi-cellular organism, or the nucleic acid is administered into a cell first, and then the cell containing the nucleic acid is administered into the body of the multi-cellular organism. After delivery into the organism, the nucleic acid may exist independently from the genome of the host organism or may be integrated into the genome of the host organism. If the nucleic acid is integrated into a germline cell of the host organism, such nucleic acid may be transmitted into the host organism's offspring.