Ultrasensitive detection of glioblastoma brain tumor cells using 5-ALA and SpectroPen for fluorescence-guided resection Público
Sharma, Abhinav K. (2014)
Abstract
SpectroPen is a handheld spectrometer that can be used intraoperatively to help surgeons detect glioblastoma tumor cells, therefore maximizing tumor cell resection by helping surgeons delineate diffuse and undefined tumor margins that are characteristic of glioblastoma. The use of a metabolic contrast agent, 5-aminolevulinic acid (5-ALA), is also tested as a means by which the SpectroPen can selectively detect fluorescence in cancer cells through the tumor cell-specific accumulation of protoporphyrin-IX (Pp-IX), a fluorescent metabolite of the 5-ALA prodrug. Here, we show that SpectroPen is capable of detecting fluorescence emission from Pp-IX in vitro with U87 EGFRvIII glioblastoma cells lines as well as from Pp-IX in tumor cells within xenograft mice models that have had U87 EGFRvIII tumor cell implantation in their brains. Additionally, the device has shown much greater sensitivity to fluorescence emission from tumor cells than the more conventional surgical fluorescent microscope, as seen by SpectroPen's detection threshold of 2,500 U87 EGFRvIII cells. My project demonstrates the efficacy of 5-ALA as a contrast agent in staining studies using U87 EGFRvIII tumor cells and normal cell lines, astrocytes and NIH/3T3, via tests for selective uptake and fluorescence in the U87 EGFRvIII cells. The results obtained from these studies are currently being translated to the clinic, where we are collaborating with Dr. Costas Hadjipanayis of Emory University Hospital Midtown. Currently, 5-ALA is not approved by the FDA in the United States and thus Dr. Hadjipanayis is currently performing Phase II clinical trials with the drug.
Table of Contents
LIST OF FIGURES ............................................................1
LIST OF TABLES .............................................................2
INTRODUCTION ...............................................................4
METHODS .....................................................................11
RESULTS/DISCUSSION.....................................................17
CONCLUSION ................................................................36
REFERENCES .................................................................39
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