WT1 Clone 6F-H2 Cytoplasmic Expression Differentiates Astrocytic Tumors from Astrogliosis and Associates with Tumor Grade, Histopathology, IDH1 Status, Apoptotic and Proliferative Indices: A Tissue Microarray Study

Abstract

Objectives: This tissue microarray (TMA) immunohistochemical (IHC) study elucidates the role of Wilms' tumor 1 protein (WT1) in diagnosis and prognostication of astrocytic tumors. Methods: IHC was applied to 75 astrocytic lesions (18 astrogliosis and 57 astrocytic tumors) using antibodies directed against WT1 clone 6F-H2, isocitrate dehydrogenase 1(IDH1), Bcl2 and Ki67. WT1 IHC staining was evaluated and scored blindly by 2 pathologists. Bcl2 and Ki67 scores and labelling indices were assessed and IDH1 status determined. Statistical analysis was performed using the appropriate methodology. Results: WT1 cytoplasmic expression was detected in 89.5% of astrocytic tumors but not in astrogliosis. Positive WT1 differentiated astrocytic tumors (92.6% accuracy) and grade II diffuse astrocytomas (93.5% accuracy) from astrogliosis with high sensitivity, specificity and positive predictive values (p<0.001). Increased WT1 score significantly associated higher Bcl2 and Ki67 labelling indices, increasing WHO tumor grade and tumor's histopathologic type (p<0.05) showing staining pattern variability by tumor entity and cell type. Glioblastomas, gliosarcomas and subependymal giant cell astrocytomas were the most frequently WT1 expressing tumors with frequent +3 WT1 score. About 21.4% of pilocytic astrocytomas had +3WT1 score in association with increased Bcl2 and Ki67 indices. Low WT1 scores in grade II and III diffuse astrocytomas were linked to the high frequency of IDH1 positivity, and were associated with low Bcl2 and Ki67 labelling indices. In glioblastomas, WT1 significantly associated poor prognostic variables: older age, negative-IDH1 status, high Bcl2 and Ki67 labelling indices (p=0.04, <0.001, =0.001 and <0.001 respectively). Conclusions: WT1 clone 6F-H2 is a highly accurate positive surrogate marker to differentiate astrocytic tumors notably the challenging grade II diffuse astrocytoma from astrogliosis. It significantly associates with poor prognostic variables including IDH1 negativity, high apoptotic and proliferative indices and depends on tumor's histopathologic entity more than its grade. Evaluation of WT1 expression seems essential to tailor patient's therapy.

Role of Wilms’ tumor 1 in diagnosis and grading of astrocytomas

“…In the current investigation, WT1 expression was identified in each of the 54 astrocytomas (100%) that were examined. Similar findings from earlier trials with positive ranging between 80.9% and 100% 25,26 have been reported.…”

“…Although the majority of WT1 molecular investigations have revealed 100% positivity, 26 several research employing the IHC technique have discovered missing expression in a small number of astrocytomas. In their analysis of 73 astrocytomas, Mahzouni and Meghdadi 27 observed that 97.3% of the time, WT1 protein was expressed.…”