PROJECT SUMMARY/ABSTRACT:
The polycomb repressive complex 2 (PRC2) complex establishes and maintains di- and tri-methylation of the
histone H3 at lysine 27 (H3K27me2/3) in the genome and regulates chromatin structure, transcription, cellular
stemness and differentiation. PRC2 is a context-dependent tumor suppressor whose core components (e.g.,
EZH2, EED, or SUZ12) are inactivated in various cancer types. Among these, high-grade malignant peripheral
nerve sheath tumor (MPNST), an aggressive soft tissue sarcoma with no effective therapies, has a high
prevalence (=80%) of biallelic inactivation of EED or SUZ12, leading to complete loss of the PRC2 function.
PRC2 loss in cancer results in aberrant transcriptional activation of developmentally silenced master regulators,
which leads to enhanced cellular plasticity and aberrant activation of multiple signaling pathways. We recently
uncovered that PRC2 loss in cancer leads to an immune-desert tumor microenvironment and resistance to
immune checkpoint blockade. Nevertheless, we observed that, in murine models of PRC2-loss MPNST and
mammary tumors, response to immunotherapy can be enhanced by infection with immunogenic virus, which
activates double-stranded RNA (dsRNA) signaling responses. Moreover, we have identified and validated a
lethal interaction between PRC2 loss and DNA methyltransferase 1 (DNMT1) knockdown in MPNST. In vitro and
in vivo treatment with a demethylating agent (decitabine) or selective DNMT1 inhibitors (GSK862, GSK032) led
to enhanced cytotoxicity and antitumor effects in PRC2-loss compared to PRC2 wild-type (wt) MPNST models.
Mechanistically, DNMT inhibitor (DNMTi) treatment in the PRC2-loss context amplified the expression of
endogenous retrotransposons and subsequently led to activation of innate immune responses, which could be
explained by retrotransposons forming dsRNA and triggering cytotoxicity via PKR-dependent dsRNA sensing.
we hypothesize that PRC2 loss in cancer may create therapeutic opportunities for agents (e.g., DNMTi, synthetic
dsRNA, immunogenic viruses) that activate the dsRNA signaling pathways in tumor cells to induce cytotoxicity
and enhance immunogenicity. Here, we propose integrative multi-omics analysis (e.g., transcriptome,
epigenome, single-cell [sc]RNA-seq and scATAC-seq) and innovative approaches (e.g., novel immunogenic
viruses, a novel lineage-tracking system) to evaluate novel therapeutic strategies and their mechanisms of
activating dsRNA responses and anti-tumor effects in relevant PRC2-loss cancers, focusing on specific aims:
(1) dissect the molecular mechanisms that underlie the selective sensitivity to DNMTi in different PRC2-loss
cancers; (2) evaluate exogenous dsRNAs as a strategy to induce cytotoxicity in PRC2-loss tumors; and (3)
evaluate novel therapeutic strategies of activating dsRNA-responses in relevant cancer models. These studies
will generate insights into exploiting dsRNA-sensing and activation of innate immune responses as therapeutic
strategies in PRC2-loss cancers and provide the pivotal preclinical data for biomarker-driven clinical trials.