Dynamics of the intratumoral immune response during progression of high-grade serous ovarian cancer

Einleitung: Das Ovarialkarzinom geht mit der höchsten Mortalität aller gynäkologischen Tumore einher. Zur schlechten Prognose tragen insbesondere die Rezidivtumore bei, die zwar einen Großteil der Patientinnen betreffen, über deren Biologie jedoch bislang nur wenig bekannt ist. Ein adäquates Therapiemanagement wird unter anderem durch eine hohe intratumorale genetische Vielfalt erschwert. Auch die anti-tumorale Immunantwort gilt als prognostischer Faktor und stellt einen möglichen Ansatzpunkt für innovative Therapiekonzepte dar. Jedoch ist die Heterogenität des Immuninfiltrates bisher nur unzureichend erforscht. Unsere Arbeit untersucht die Dynamik des Immuninfiltrats während der Tumorprogression erstmals in gepaarten Primär- und Rezidivtumoren des high-grade serösen Ovarialkarzinoms (HGSOC). Methodik: Wir untersuchten die Dichte CD3+, CD4+, CD8+ tumorinfiltrierender Lymphozyten (TILs) und MHC1, 2 Expression in 113 gepaarten primären und rezidivierten HGSOCs immunhistochemisch an Tissue Microarrays (TMAs). Die TILs Quantifizierung erfolgte erstmals über eine neue, automatisierte Bildanalysesoftware (CD3 Quantifier), die im Rahmen dieser Arbeit validiert und weiterentwickelt wurde. Alle Patientinnen waren Teil des EU-finanzierten OCTIPS FP7 Projekts. Ergebnisse: Wir fanden eine signifikante Korrelation der TILs Dichte und MHC Expression zwischen Primär- und Rezidivtumor. Die anti-tumorale Immunantwort zeigte sich im Verlauf stabil, jedoch ergaben sich in einigen Tumoren Hinweise auf eine verstärkte Immunantwort im Rezidiv. Die meisten immunologischen Marker verloren im Rezidivtumor ihren prognostischen Wert. Schlussfolgerung: Die aufgeführten Ergebnisse zeigen eine homogene Immunantwort im Verlauf des HGSOC erstmals im Vergleich zwischen Primär- und Rezidivtumor. Dass eine erhöhte Immuninfiltration im Rezidiv nicht mit einer verbesserten Prognose einhergeht, lässt zudem vermuten, dass eine Effektivität der anti-tumoralen Immunantwort im Laufe der Tumorprogression nicht zwingend gegeben ist. Unsere Arbeit zeigt, dass eine Gewebeprobe des Primärtumors auch für die immunologische Untersuchung des Rezidivs verwendet werden könnte und somit Rückschlüsse auf das bislang nur unzureichend erforschte rezidivierte Ovarialkarzinom zulässt.Introduction: Epithelial ovarian cancer (EOC) is associated with the highest mortality rate among all gynecological cancer. Poor prognosis is especially attributed to recurrent disease, which a majority of women develop, but which to date is only poorly understood. To make matters worse, EOC is known for a high intratumoral genetic variability that impedes an adequate therapeutic management. Apart from that, the anti-tumoral immune response is perceived as a pivotal prognostic factor and a potential base for immunotherapy. Still, research on the heterogeneity of intratumoral immunoinfiltration remains scant. In this study we analyze the dynamic of the intratumoral immune response during progression of high-grade serous ovarian cancer (HGSOC) and therefore compare paired tumor samples of primary and recurrent disease for the first time. Methods: We systematically analyzed the densities of CD3+, CD4+, CD8+ tumorinfiltrating lymphocytes (TILs) and MHC1, 2 expression by immunohistochemistry on tissue microarrays (TMAs) in 113 paired samples of primary and recurrent HGSOC. TILs quantifying was performed using a new automated software (CD3 Quantifier) for the first time. As part of this study the software was validated and further developed by comparative analyses. All patients had been included to the EU-funded OCTIPS FP7 project. Results: Our analysis revealed a correlation of TILs density and MHC expression between primary and recurrent HGSOC. The anti-tumoral immune response remained stable during tumor progression, however, part of the tumors showed an enhanced immunogenicity in recurrent tumors. Most of the immunological markers lost their prognostic impact in recurrent disease. Conclusion: We are the first ones to show a homogenous intratumoral immune response across primary and recurrent tumors in HGSOC. A higher anti-tumoral immune response was not associated with a better prognosis in recurrent tumor. Thus, we assume an immune response during tumor progression might not necessarily be effective. Our findings suggest that tissue samples of a primary lesion could also be used for the immunological analysis of the recurrent tumor. This would allow to gain further insights into the investigation of the so far poorly understood recurrent EOC

Exploring the clonal evolution of CD133/aldehyde-dehydrogenase-1 (ALDH1)-positive cancer stem-like cells from primary to recurrent high-grade serous ovarian cancer (HGSOC). A study of the Ovarian Cancer Therapy–Innovative Models Prolong Survival (OCTIPS) Consortium

Background High-grade serous ovarian cancer (HGSOC) causes 80% of all ovarian cancer (OC) deaths. In this setting, the role of cancer stem-like cells (CSCs) is still unclear. In particular, the evolution of CSC biomarkers from primary (pOC) to recurrent (rOC) HGSOCs is unknown. Aim of this study was to investigate changes in CD133 and aldehyde dehydrogenase-1 (ALDH1) CSC biomarker expression in pOC and rOC HGSOCs. Methods Two-hundred and twenty-four pOC and rOC intrapatient paired tissue samples derived from 112 HGSOC patients were evaluated for CD133 and ALDH1 expression using immunohistochemistry (IHC); pOCs and rOCs were compared for CD133 and/or ALDH1 levels. Expression profiles were also correlated with patients' clinicopathological and survival data. Results Some 49.1% of the patient population (55/112) and 37.5% (42/112) pOCs were CD133+ and ALDH1+ respectively. CD133+ and ALDH1+ samples were detected in 33.9% (38/112) and 36.6% (41/112) rOCs. CD133/ALDH1 coexpression was observed in 23.2% (26/112) and 15.2% (17/112) of pOCs and rOCs respectively. Pairwise analysis showed a significant shift of CD133 staining from higher (pOCs) to lower expression levels (rOCs) (p < 0.0001). Furthermore, all CD133 + pOC patients were International Federation of Gynaecology and Obstetrics (FIGO)-stage III/IV (p < 0.0001) and had significantly worse progression-free interval (PFI) (p = 0.04) and overall survival (OS) (p = 0.02). On multivariate analysis, CD133/ALDH1 coexpression in pOCs was identified as independent prognostic factor for PFI (HR: 1.64; 95% CI: 1.03–2.60; p = 0.036) and OS (HR: 1.71; 95% CI: 1.01–2.88; p = 0.045). Analysis on 52 pts patients with known somatic BRCA status revealed that BRCA mutations did not influence CSC biomarker expression. Conclusions The study showed that CD133/ALDH1 expression impacts HGSOC patients' survival and first suggests that CSCs might undergo phenotypic change during the disease course similarly to non stem-like cancer cells, providing also a first evidence that there is no correlation between CSCs and BRCA status

Characterisation of tumor microvessel density during progression of high-grade serous ovarian cancer: clinico-pathological impact. An OCTIPS Consortium study.

Background: High-grade serous ovarian cancer (HGSOC) intratumoural vasculature evolution remains unknown. The study investigated changes in tumour microvessel density (MVD) in a large cohort of paired primary and recurrent HGSOC tissue samples and its impact on patients’ clinico-pathological outcome. Methods: A total of 222 primary (pOC) and recurrent (rOC) intra-patient paired HGSOC were assessed for immunohistochemical expression of angiogenesis-associated biomarkers (CD31, to evaluate MVD, and VEGF-A). Expression profiles were compared between pOCs and rOCs and correlated with patients' data. Results: High intratumoural MVD and VEGF-A expression were observed in 75.7% (84/111) and 20.7% (23/111) pOCs, respectively. MVDhighand VEGF(+)samples were detected in 51.4% (57/111) and 20.7% (23/111) rOCs, respectively. MVDhigh/VEGF(+)co-expression was found in 19.8% (22/111) and 8.1% (9/111) of pOCs and rOCs, respectively (p = 0.02). Pairwise analysis showed no significant change in MVD (p = 0.935) and VEGF-A (p = 0.121) levels from pOCs to rOCs. MVDhighpOCs were associated with higher CD3(+)(p = 0.029) and CD8(+)(p = 0.013) intratumoural effector TILs, while VEGF(+)samples were most frequently encountered among BRCA-mutated tumours (p = 0.019). Multivariate analysis showed VEGF and MVD were not independent prognostic factors for OS. Conclusions: HGSOC intratumoural vasculature did not undergo significant changes during disease progression. High concentration of CD31(+)vessels seems to promote recruitment of effector TILs. The study also provides preliminary evidence of the correlation between VEGF-positivity and BRCA status