Abstract A082: The epigenetic impact and therapeutic opportunity of AR-directed therapy for desmoplastic small round cell tumor
Authors: Truong Danh D. , Lamhamedi-Cherradi Salah-Eddine , Maitituoheti Mayinuer , Beird Hannah C. , Wu Chia-Chin , Krishnan Sandhya , Ingram Davis , Futreal P. Andrew , Titus Mark , Lazar Alexander , Rai Kunal , MacLeod A. Robert , Daw Najat C. , Hayes Andrea , Ludwig Joseph 
Published: June 2023

Abstract
Desmoplastic small round cell tumor (DSRCT) is a rare, usually incurable, aggressive sarcoma subtype that occurs predominantly in adolescent and young adult (AYA) males. At diagnosis, most present with hundreds of intraabdominal nodules composed of malignant cells that harbor a EWSR1-WT1 chromosomal translocation and resultant fusion protein (FP). It was shown that a subset of DSRCT cells (mostly of epithelial histotype) highly express the androgen receptor (AR), a key epigenetic driver of prostate cancer (PC). Given the male predilection and the high expression of AR, we sought to assess the role of AR more comprehensively in DSRCT and ask if this tumor shared AR resistance mechanism to PC. We tested if AR antagonists curb cell proliferation and tumor growth, where both enzalutamide and AR-antisense therapy blocked DHT-induced cell proliferation and reduced xenograft tumor burden. Next, to mechanistically interrogate AR’s oncogenic effects, we performed single-nuclei RNA-seq (snRNA-eq) and chromatin immunoprecipitation sequencing (ChIP-seq) on frozen patient specimens. Those studies revealed a surprising epigenetic similarity between DSRCT to PC. Though overlap exists in the DNA binding MOTIFs of AR in DSRCT and PC, our ChIP-seq resultsrevealed DSRCT-specific AR DNA binding sites adjacent to key oncogenic regulators, including FOXF1 and WT1 (the C-terminal partner of the pathognomonic FP). To our surprise, we identified a subset of DSRCT samples that underwent partial neuroendocrine (NE) epigenetic reprogramming, akin to what occurs in ~1/3 of castrate-resistant PC (CRPC). Since androgen deprivation therapy (ADT) isn’t yet a standard treatment for DSRCT, it remains an enigma why DSRCT would undergo NE reprogramming or how this may affect ADT sensitivity. To address this, nine DSRCT patient samples were profiled using snRNA-seq. Sub-clustering revealed epithelial, mesenchymal, or NE signatures and marked inter-patient and intra-tumoral heterogeneity. Notably, five of nine patients exhibited NE markers (SYP, ENO2, CHGA, FOXA2, ASCL1, and SOX2), while three expressed epithelial markers (MUC1, MUC6, KRT18, KRT23, CDH1). One patient exhibited a hybrid AR indifferent phenotype. While strikingly different from PC morphologically and phenotypically, our data suggest that DSRCT is a second androgen-stimulated malignancy (third if we consider breast cancers that rely on shared nuclear receptor family signaling). Shared dependence upon AR for tumor growth and survival provides an exciting opportunity to prospectively study AR signaling in a different cancer type and younger DSRCT-stricken patient population. Ongoing work will determine if DSRCT undergoes dedifferentiation towards a more stem-like cell as an intermediary step before undergoing neural lineage commitment or transdifferentiation directly from an epithelial cell type towards a neuroendocrine cell fate. We are also exploring if the FP acts as a Pioneer factor to direct AR towards DSRCT-specific androgen response elements that explain this sarcoma subtype’s unique clinical presentation.
Citation Format: Danh D. Truong, Salah-Eddine Lamhamedi-Cherradi, Mayinuer Maitituoheti, Hannah C. Beird, Chia-Chin Wu, Sandhya Krishnan, Davis Ingram, P. Andrew Futreal, Mark Titus, Alexander Lazar, Kunal Rai, A. Robert MacLeod, Najat C. Daw, Andrea Hayes, Joseph Ludwig. The epigenetic impact and therapeutic opportunity of AR-directed therapy for desmoplastic small round cell tumor [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A082.