Rare-06. dissecting cellular differentiation heterogeneity in choroid plexus carcinoma pathogenesis

Abstract Choroid plexus (CP) neoplasms are rare, predominantly pediatric, brain tumors ranging from benign CP papilloma to aggressive CP carcinoma (CPC) associated with poor survival. Survivors often experience devastating side effects from current treatment strategies. There is an urgent need for safer, more effective therapies for CPC. A subset of human CPC exhibits abnormal NOTCH activity, whereas mutations of tumor suppressor gene TP53 are detected in >50% of CPC and are associated with increased genetic tumor instability and worse prognosis. CPC is one hallmark of Li-Fraumeni syndrome (LFS), a rare genetic disorder associated with germline TP53 mutations. Human CP tumors also frequently display aberrant expression of transcriptional regulators of roof plate/CP differentiation, including LIM homeodomain transcription factors LMX1A, and SRY-Box Transcription Factor 2 (SOX2). To investigate the role of transcriptional regulation in CP tumorigenesis, animal models were developed driven by molecular defects commonly found in CP tumors in humans. Accordingly, sustained NOTCH and Sonic Hedgehog (SHH) activation or combined deletion of Rb1 and Trp53 tumor suppressors led to malignant CP tumors with characteristics of CPC in humans. NOTCH-driven CP tumors arose from monociliated roof plate progenitors and exhibited increased levels of Lmx1a, Lmx1b, and Sox2. In contrast, their expression was significantly reduced, while OTX2 levels were markedly increased in Rb1/Trp53-deficient CPC. Consistently, single-cell transcriptomics data show a proliferative Otx2+ tumor bulk and a small Lmx1a+/Sox2+ population in Rb1/Trp53-deficient CPC. Importantly, deletion of Sox2 in Lmx1a+ progenitors blocked NOTCH-driven CP tumor and reduced Lmx1a/b expression. Thus, LMX1A and SOX2 expression marks tumor-initiating cells, while OTX2 labels differentiated tumor cells in CPC. Together, our studies revealed intra- and inter-tumoral heterogeneity in CPC driven by distinct oncogenic signals. Knowledge of the molecular and cellular basis of heterogeneity in CP tumor development may facilitate the development of innovative diagnostic and therapeutic strategies in CPC.