TY - UNPB
T1 - Localization of protoporphyrin IX in glioma patients with paired stimulated Raman histology and two-photon excitation fluorescence microscopy
AU - Nasir-Moin, Mustafa
AU - Wadiura, Lisa
AU - Juros, Devin
AU - Movahed-Ezazi, Misha
AU - Lee, Matthew
AU - Weiss, Hannah
AU - Müther, Michael
AU - Alber, Daniel
AU - Ratna, Sujay
AU - Fang, Camila
AU - Suero-Molina, Eric
AU - Hellwig, Sonke
AU - Stummer, Walter
AU - Rössler, Karl
AU - Hainfellner, Johannes
AU - Widhalm, Georg
AU - Kiesel, Barbara
AU - Reichert, David
AU - Mischkulnig, Marlo
AU - Jain, Rajan
AU - Smith, Andrew
AU - Straehle, Jakob
AU - Neidert, Nicholas
AU - Schnell, Oliver
AU - Beck, Jurgen
AU - Trautman, Jay
AU - Pastore, Steve
AU - Pacione, Donato
AU - Placantonakis, Dimitris
AU - Oermann, Eric
AU - Golfinos, John
AU - Hollon, Todd
AU - Snuderl, Matija
AU - Freudiger, Christian
AU - Heiland, Dieter Henrik
AU - Orringer, D.
PY - 2022
Y1 - 2022
N2 - Fluorescence guidance is widely utilized to improve the precision of cancer surgery. 5-aminolevulinic acid, the most widely used fluorophore in glioma surgery, is thought to cause selective accumulation of fluorescent protoporphyrin IX (PpIX) in tumor cells. 5-aminolevulinic acid is highly specific for densely tumor-infiltrated tissue but less effective for visualizing the tumor periphery. To improve clinical detection of PpIX, we developed a microscope to perform paired stimulated Raman histology and two-photon excitation fluorescence microscopy (TPEF) and validated it in 175 fresh tumor specimens from 75 high-grade glioma patients across three institutions. Here, we demonstrate that intracellular PpIX accumulation occurs most prominently in histiocytic, rather than neoplastic, appearing cells. Spatially resolved metabolomics, transcriptomics and RNA sequencing revealed that PPIX is most avidly concentrated in tumor associated macrophages. There was no correlation between the degree of tissue cellularity and PpIX concentration across all imaged specimens (R=-0.21). Our findings encourage reconsideration of the existing theory of 5-ALA-induced glioma cell fluorescence and demonstrate how 5-ALA and TPEF imaging can provide a window into the immune microenvironment of human gliomas.
AB - Fluorescence guidance is widely utilized to improve the precision of cancer surgery. 5-aminolevulinic acid, the most widely used fluorophore in glioma surgery, is thought to cause selective accumulation of fluorescent protoporphyrin IX (PpIX) in tumor cells. 5-aminolevulinic acid is highly specific for densely tumor-infiltrated tissue but less effective for visualizing the tumor periphery. To improve clinical detection of PpIX, we developed a microscope to perform paired stimulated Raman histology and two-photon excitation fluorescence microscopy (TPEF) and validated it in 175 fresh tumor specimens from 75 high-grade glioma patients across three institutions. Here, we demonstrate that intracellular PpIX accumulation occurs most prominently in histiocytic, rather than neoplastic, appearing cells. Spatially resolved metabolomics, transcriptomics and RNA sequencing revealed that PPIX is most avidly concentrated in tumor associated macrophages. There was no correlation between the degree of tissue cellularity and PpIX concentration across all imaged specimens (R=-0.21). Our findings encourage reconsideration of the existing theory of 5-ALA-induced glioma cell fluorescence and demonstrate how 5-ALA and TPEF imaging can provide a window into the immune microenvironment of human gliomas.
UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-85146094518&partnerID=MN8TOARS
U2 - 10.21203/rs.3.rs-1519287/v1
DO - 10.21203/rs.3.rs-1519287/v1
M3 - Preprint
T3 - Research Square
BT - Localization of protoporphyrin IX in glioma patients with paired stimulated Raman histology and two-photon excitation fluorescence microscopy
ER -