![The oncogenic role of STRN3-RARA fusion gene in leukemogenesis. (A) Treatment curve of U937 cells with ATRA and ATO. Cells were transduced with vector or STRN3-RARA. Means ± standard deviation (SD), n = 3; ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001 (2-way analysis of variance [ANOVA]). (B) Western blot analysis of ATRA-treated 293T cells harvested 2 days after treatment. 293T cells were transduced with Flag-STRN3-RARA. (C) GSEA showing positive enrichment of the APL_PML_RARA gene set in U937 cell with STRN3-RARA overexpression, compared with vector (NES = 1.57; P = .01). (D) Levels of STRN3-RARA bound at the transcription start site (TSS) in U937 cells transduced with STRN3-RARA, measured by the CUT&Tag analysis. (E) The Venn diagram showing the overlap of binding-genes in STRN3-RARA and PML-RARA (hypergeometric test). (F) Kaplan-Meier tumor-free survival curves of recipient mice in vector and STRN3-RARA group with Utx knockout, n = 6 (vector), n = 8 (STRN3-RARA); ∗∗P < .01 (log-rank test). (G) Blood smear of sgUtx;SR recipient mouse before sacrificed. Scale bar, 10 μm. (H) The percentage of sgUtx;SR cells (GFP + mCherry + cells) along with vehicle and ATRA treatment. The x-axis indicates the number of days after treatment. Means ± SD, vehicle, n = 5, 4, 4, 2; ATRA, n = 5, 4, 4, 4; ∗∗∗∗P < .0001 (2-way ANOVA). (I) Blood smears from sgUtx;SR mice following a 25-day treatment with vehicle or ATRA. Scale bar, 10 μm. (J) Survival curve of sgUtx;SR mice following a 25-day ATRA or vehicle treatment. n = 5; ∗P < .05 (log-rank test). (K) Heat map showing the 140 drug targetable genes that were specifically upregulated in U937 cells with sgUTX;SR compared with that with empty vectors and not overlapped with the upregulated genes due to PML-RARA. (L) GSEA showing the positive enrichment of the HALLMARK_TNFA_SIGNALING_VIA_NFKB gene set in sgUTX;SR U937 cells, compared with vector (NES = 1.67; P = .00). (M) Relative viability of U937 cells treated with 1 μM cepharanthine. Vector, sgUTX, SR, and sgUTX;SR groups are shown. Means ± SD, n = 3; ∗P < .05, ∗∗P < .01 (1-way ANOVA). (N) Drug treatment survival curve of WCH01 relapsed cells. ATRA and cepharanthine were treated respectively. Means ± SD, n = 3; ∗∗P < .01, ∗∗∗P < .001 (2-way ANOVA). IgG, immunoglobulin G; Veh, vehicle.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/142/17/10.1182_blood.2023020619/2/blood_bld-2023-020619-gr2b.jpeg?Expires=1735558483&Signature=D~rXVqrCTNdMKLMqEhZdkle85xbNapxYvMNtWYVBUT7oMWF~tinxjZ7MQNnkW0HLG71gwSCNGbvnecROWk0BlAtMsO0M-LQzprXrY7AxXgtWhvfPfPNn2xzQ0DFumobIqf0ubKpVuuaKFfhZQQ-xUWjyXMkw6Ms8ajHlwxJy2RtKJ1HlAzXdnXsu7RmvTaShRFEyQ8la-v0bop~zJ3m1Wha0oxrZdrlEG4F8ndtbqs6oJWE1Ut2PWeZfesPKrDEHmEsKf8QZWqAYf4o65b4Ma0OK28LXgKxiIuRSg0i7-b30qnFpnwtxCyGQYwsG69cuw788wr5i2rMFojAdo0XZVQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
The oncogenic role of STRN3-RARA fusion gene in leukemogenesis. (A) Treatment curve of U937 cells with ATRA and ATO. Cells were transduced with vector or STRN3-RARA. Means ± standard deviation (SD), n = 3; ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001 (2-way analysis of variance [ANOVA]). (B) Western blot analysis of ATRA-treated 293T cells harvested 2 days after treatment. 293T cells were transduced with Flag-STRN3-RARA. (C) GSEA showing positive enrichment of the APL_PML_RARA gene set in U937 cell with STRN3-RARA overexpression, compared with vector (NES = 1.57; P = .01). (D) Levels of STRN3-RARA bound at the transcription start site (TSS) in U937 cells transduced with STRN3-RARA, measured by the CUT&Tag analysis. (E) The Venn diagram showing the overlap of binding-genes in STRN3-RARA and PML-RARA (hypergeometric test). (F) Kaplan-Meier tumor-free survival curves of recipient mice in vector and STRN3-RARA group with Utx knockout, n = 6 (vector), n = 8 (STRN3-RARA); ∗∗P < .01 (log-rank test). (G) Blood smear of sgUtx;SR recipient mouse before sacrificed. Scale bar, 10 μm. (H) The percentage of sgUtx;SR cells (GFP + mCherry + cells) along with vehicle and ATRA treatment. The x-axis indicates the number of days after treatment. Means ± SD, vehicle, n = 5, 4, 4, 2; ATRA, n = 5, 4, 4, 4; ∗∗∗∗P < .0001 (2-way ANOVA). (I) Blood smears from sgUtx;SR mice following a 25-day treatment with vehicle or ATRA. Scale bar, 10 μm. (J) Survival curve of sgUtx;SR mice following a 25-day ATRA or vehicle treatment. n = 5; ∗P < .05 (log-rank test). (K) Heat map showing the 140 drug targetable genes that were specifically upregulated in U937 cells with sgUTX;SR compared with that with empty vectors and not overlapped with the upregulated genes due to PML-RARA. (L) GSEA showing the positive enrichment of the HALLMARK_TNFA_SIGNALING_VIA_NFKB gene set in sgUTX;SR U937 cells, compared with vector (NES = 1.67; P = .00). (M) Relative viability of U937 cells treated with 1 μM cepharanthine. Vector, sgUTX, SR, and sgUTX;SR groups are shown. Means ± SD, n = 3; ∗P < .05, ∗∗P < .01 (1-way ANOVA). (N) Drug treatment survival curve of WCH01 relapsed cells. ATRA and cepharanthine were treated respectively. Means ± SD, n = 3; ∗∗P < .01, ∗∗∗P < .001 (2-way ANOVA). IgG, immunoglobulin G; Veh, vehicle.
