Digital Image Analysis using Qupath to determine immune cell content in formalin-fixed, paraffin-embedded murine neuroblastoma tumors

Sammanfattning: Neuroblastoma (NB), an extracranical solid tumor, is among the most prevalent cancers affecting children, particularly those under the age of five. High-risk NB presents a survival rate just below 50 %. Angiogenesis, a crucial process in NB, is induced by various pro-angiogenic factors. The compound SU11657 has demonstrated efficacy in inhibiting angiogenesis and tumor progression. Tumor-associated macrophages (TAMs) and Tumor-associated neutrophils (TANs) contribute to tumor progression, including angiogenesis, and their heightened levels within the tumor has been correlated with a poor clinical prognosis. This study aimed to quantify TANs and TAMs in NB tumors through manual assessment and the development of an automated digital image analysis. Unfortunately, due to time constraints, TAMs were not subjected to detailed analysis. Immunohistochemistry using antibody ab2557 and DAB staining was employed, and cell content analysis was performed through both manual assessment and digital analysis using QuPath. Successful differentiation of TANs was achieved with ab2557. The manual assessment observed a decrease of TANs between the control and treatment groups in UB7 and UB8, with UB7 being statistically significant (p<0.05), based on a two-tailed t-test. QuPath analysis noted increases in the percentages of TANs between the control and treatment groups, with the t-tests being non-significant (p>0.05). While digital image analysis is gaining importance in clinical applications, imperfections persist, underscoring the imperative for further research and development to accurately distinguish biomarkers.