Session: 622. Lymphomas: Translational – Non-Genetic: Poster I
Hematology Disease Topics & Pathways:
Research, Translational Research
Salivary gland NK cells are tissue-resident NK cells that are distinct from conventional NK cells. We investigated how Trp53 deficiency has different effects on NK cells across these tissues in the pre-tumor state. Remarkably, Trp53 knockout caused an increase of Lin-CD122+NK1.1- cells and extensive gene expression changes of NK cells exclusively in the salivary glands. Thus, we evaluated characteristic phenotypes of tissue-resident NK cells and found that not only murine NK-cell tumors but also human ENKTCL samples expressed tissue-resident markers such as CD69 and CD49a, suggesting tissue-resident NK cells as their cell-of-origin. To investigate genetic profiles of Trp53-/- tumors, we performed whole-exome sequencing, which revealed recurrent Myc amplifications. In addition, RNA sequencing (RNA-seq) showed the significant enrichment of MYC target gene signatures in these tumors.
To investigate the role of EBV-encoded latent membrane protein 1 (LMP1) in NK-cell lymphomagenesis, we introduced the expression of LMP1 into our mouse model (Trp53-/-LMP1+). LMP1 expression significantly accelerated NK-cell lymphomagenesis. Single-cell RNA-seq of the tumors showed LMP1-mediated propagation of myeloid cells, including monocytes and conventional dendritic cells (DC), through interferon-γ signaling. Cell-cell interaction analysis revealed that LMP1-induced myeloid cell expansion promoted tumor cell proliferation through upregulated CXCL16-CXCR6 signaling. We validated the presence of CD11c+ DC in the tumor microenvironment and CXCR6 and CXCR3 protein expressions in tumor cells by immunohistochemical analysis of human ENKTCL samples. Importantly, inhibition of CXCL16-CXCR6 signaling was effective against NK-cell tumors in vivo.
Using single-cell RNA-seq and flow cytometric analyses, we also found the significant expansion of KLRG1-expressing cells within the tumors. These cells harbored the capacity to repopulate tumors in secondary recipients. We validated the upregulation of KLRG1 expression in human ENKTCL samples through RNA-seq data and immunohistochemical analysis. We then examined whether targeting tumor cells with anti-KLRG1 antibody is effective against murine NK-cell tumors and found that anti-KLRG1 antibody strikingly prolonged the survival of mice transplanted with Trp53-/- tumors. As the effectiveness of anti-KLRG1 antibody was modest against Trp53-/-LMP1+ tumors, we further explored another therapeutic target and identified further upregulation of MYC target gene signature in Trp53-/-LMP1+ tumors. Therefore, we treated mice bearing Trp53-/-LMP1+ tumors with silvestrol, an eIF4A inhibitor that blocks Myc translation. While silvestrol alone slightly improved the survival, combining it with an anti-KLRG1 antibody further extended the survival.
In conclusion, using GEMM recapitulating the human disease, we have delineated the cell-of-origin and microenvironmental changes and exploited novel therapeutic targets, including CXCL16, KLRG1, and MYC for ENKTCL. These observations prove that our mouse models will serve as valuable tools for elucidating the pathogenic mechanisms and refining diagnostic and therapeutic strategies in ENKTCL.
Disclosures: Kogure: Kyowa Kirin: Honoraria; Nippon Shinyaku: Honoraria; Takeda Pharmaceutical: Honoraria; Daiichi Sankyo: Honoraria. Chaubard: Correspondances en Onco-Hématologie: Research Funding. Jaccard: Jazz Pharmacueticals: Honoraria; janssen: Honoraria; sanofi: Research Funding; pfizer: Honoraria. Hermine: Roche: Research Funding; Alexion: Research Funding; BMS: Research Funding; Inatherys: Consultancy, Current equity holder in publicly-traded company, Patents & Royalties, Research Funding; AB Science: Consultancy, Current equity holder in publicly-traded company, Patents & Royalties, Research Funding; MSD Avenir: Research Funding. Gaulard: Gilead: Honoraria, Research Funding; Takeda Pharmaceutical: Consultancy, Honoraria, Research Funding; Innate Pharma: Research Funding; Alderan: Research Funding; Sanofi: Research Funding. Ohshima: Bristol-Myers Squibb: Research Funding; Chugai: Honoraria, Research Funding; Daiichi Sankyo: Honoraria, Research Funding; Kyowa Kirin: Research Funding. Kataoka: Celgene: Honoraria; Ono Pharmaceutical: Honoraria, Research Funding; Eisai: Honoraria, Research Funding; Chordia Therapeutics: Research Funding; Kyowa Kirin: Honoraria, Research Funding; Meiji Seika Pharma: Honoraria; Pfizer: Honoraria; AstraZeneca: Honoraria; SymBio Pharmaceuticals: Honoraria; Otsuka Pharmaceutical: Honoraria, Research Funding; Japan Blood Products Organization: Research Funding; Mochida Pharmaceutical: Research Funding; Takeda Pharmaceutical: Honoraria, Research Funding; Shionogi: Research Funding; Teijin Pharma: Research Funding; Nippon Shinyaku: Honoraria; Alexion Pharmaceuticals: Honoraria; Asahi Genomics: Current equity holder in private company; JCR Pharmaceuticals: Research Funding; Sumitomo Dainippon Pharma: Honoraria, Research Funding; Chugai Pharmaceutical: Honoraria, Research Funding; AbbVie: Honoraria; Sanofi: Honoraria; Daiichi Sankyo: Honoraria; Novartis: Honoraria; Asahi Kasei Pharma: Research Funding; Bristol-Myers Squibb: Honoraria; Janssen Pharmaceutical: Honoraria.
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