title: M2/M1 ratio of tumor associated macrophages and PPAR-gamma expression in uveal melanomas with class 1 and class 2 molecular profiles.
abstract: Macrophages have been found to be negative predictors of outcome in patients with uveal melanoma. In particular, recent studies point toward a disease-progressing role of proangiogenic M2 macrophages in melanomas with monosomy 3. Although most studies implicate a protective effect of PPAR-gamma activation in tumors, PPAR-gamma has also been shown to promote the polarization of M1 macrophages toward the M2 phenotype. The purpose of this investigation was first, to characterize the phenotype of tumor infiltrating macrophages and second, to study PPAR-gamma expression in uveal melanomas with molecular gene expression profile as prognostic predictors for patients' outcome. Twenty specimens from patients with uveal melanoma were analyzed for clinical and histologic tumor characteristics. The molecular RNA profile (class 1 or class 2) was commercially determined. Using immunohistochemical techniques, the specimens were dual labeled for CD68 and CD163. CD68 + CD163- M1 macrophages and CD68 + CD163+ M2 macrophages were analyzed in ten high power fields sparing macrophage-poor areas and a mean value was calculated for each tumor. The tumors were immunostained for von Willebrand factor and the micro vascular density (MVD) was analyzed according to Foss. To assess the proliferative rate of each tumor, Ki67 expression was evaluated in ten high power fields followed by calculation of a mean value. Expression of PPAR-gamma was evaluated using a score from 0 (no staining) to 3 (tumor entirely stained). Statistical analysis and a respective correlation were made between histologic characteristics, molecular profile, type of tumor infiltrating macrophages (M1 vs. M2), MVD, proliferative rate, and PPAR-gamma expression. Our results showed a correlation between the ratio of M2/M1 macrophages and the molecular profile with a ratio of approximately 1 corresponding to molecular class 1 and a ratio of approximately 2 corresponding to molecular class 2 (p = 0.01). The ratio of M2/M1 macrophages was higher in tumors with extraocular extension (p = 0.01). PPAR-gamma was predominantly expressed in the cytoplasm of tumor cells. Its expression showed no association with the molecular RNA profile (p = 0.83). This study confirmed that the ratio of M2/M1 macrophages is another prognostic factor in uveal melanoma. Thus, polarization of macrophages plays an important role for patients' outcome. PPAR-gamma is expressed in uveal melanoma tumor cells and further studies are warranted to determine its role in tumor biology.
authors.affiliation: Dept. of Ophthalmology, Emory University, 1365 Clifton Road, BT 428, Atlanta, GA 30322, USA. martina.herwig@ukb.uni-bonn.de
authors.forename: Martina C
authors.initials: MC
authors.lastname: Herwig
authors.forename: Chris
authors.initials: C
authors.lastname: Bergstrom
authors.forename: Jill R
authors.initials: JR
authors.lastname: Wells
authors.forename: Tobias
authors.initials: T
authors.lastname: Höller
authors.forename: Hans E
authors.initials: HE
authors.lastname: Grossniklaus
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D047495:PPAR gamma / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000378:metabolism; D014604:Uveal Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D055815:Young Adult; D014841:von Willebrand Factor / Q000378:metabolism; D000098902:CD163 Antigen; D000098943:Uveal Melanoma; D000097382:CD68 Molecule
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D019394:Ki-67 Antigen; D047495:PPAR gamma; D012334:RNA, Neoplasm; D011956:Receptors, Cell Surface; D014841:von Willebrand Factor; D000098902:CD163 Antigen; C067980:CD68 antigen, human; D000097382:CD68 Molecule