Translational regulation of SND1 governs endothelial homeostasis during stress
Zhenbo Han, … , Soroush Tahmasebi, Sang-Ging Ong
Zhenbo Han, … , Soroush Tahmasebi, Sang-Ging Ong
Published February 3, 2025
Citation Information: J Clin Invest. 2025;135(3):e168730. https://doi.org/10.1172/JCI168730.
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Research Article Vascular biology

Translational regulation of SND1 governs endothelial homeostasis during stress

Abstract

Translational control shapes the proteome and is particularly important in regulating gene expression under stress. A key source of endothelial stress is treatment with tyrosine kinase inhibitors (TKIs), which lowers cancer mortality but increases cardiovascular mortality. Using a human induced pluripotent stem cell–derived endothelial cell (hiPSC-EC) model of sunitinib-induced vascular dysfunction combined with ribosome profiling, we assessed the role of translational control in hiPSC-ECs in response to stress. We identified staphylococcal nuclease and tudor domain–containing protein 1 (SND1) as a sunitinib-dependent translationally repressed gene. SND1 translational repression was mediated by the mTORC1/4E-BP1 pathway. SND1 inhibition led to endothelial dysfunction, whereas SND1 OE protected against sunitinib-induced endothelial dysfunction. Mechanistically, SND1 transcriptionally regulated UBE2N, an E2-conjugating enzyme that mediates K63-linked ubiquitination. UBE2N along with the E3 ligases RNF8 and RNF168 regulated the DNA damage repair response pathway to mitigate the deleterious effects of sunitinib. In silico analysis of FDA-approved drugs led to the identification of an ACE inhibitor, ramipril, that protected against sunitinib-induced vascular dysfunction in vitro and in vivo, all while preserving the efficacy of cancer therapy. Our study established a central role for translational control of SND1 in sunitinib-induced endothelial dysfunction that could potentially be therapeutically targeted to reduce sunitinib-induced vascular toxicity.

Authors

Zhenbo Han, Gege Yan, Jordan Jousma, Sarath Babu Nukala, Mehdi Amiri, Stephen Kiniry, Negar Tabatabaei, Youjeong Kwon, Sen Zhang, Jalees Rehman, Sandra Pinho, Sang-Bing Ong, Pavel V. Baranov, Soroush Tahmasebi, Sang-Ging Ong

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Figure 2

Ribo-Seq of hiPSC-ECs identified that sunitinib inhibits SND1 translation.

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Ribo-Seq of hiPSC-ECs identified that sunitinib inhibits SND1 translatio...
(A) Schematic of RNA-Seq and Ribo-Seq of hiPSC-ECs to identify translationally regulated mRNAs in response to sunitinib treatment. Two independent hiPSC-EC lines were used for these analyses. (B) Read-length distribution and triplet periodicity for the Ribo-Seq dataset generated by RNase I. The reads are from a single principal transcript isoform from each gene. CDS, coding sequence. (C) Percentage of raw RNA-Seq and Ribo-Seq read counts over mRNA functional regions. C, control; S, sunitinib. (D) Metagene profile of normalized Ribo-Seq read density at the corresponding base positions relative to the start and stop codons. (E) Calculated translational efficiency (TE) of genes in sunitinib-treated (2 μM) versus DMSO-treated cells. Genes that did not change significantly are colored gray; upregulated genes are in blue; and downregulated genes are in red. (F) Normalized ribosome footprint (Ribo-Seq) reads and RNA-Seq reads of SND1 in response to DMSO or sunitinib treatment were analyzed by DESeq2. (G) RT-qPCR analysis of SND1 expression in hiPSC-ECs following sunitinib treatment revealed that the mRNA level of SND1 was unaltered. Two-tailed Student’s t test. Data are presented as mean ± SD. n = 9 replicates from the differentiation of 3 individual hiPSC lines. (H and I) Immunoblot analysis and quantification of SND1 expression in hiPSC-ECs in response to DMSO or sunitinib treatment. Two-tailed Student’s t test. Data are presented as mean ± SD. ***P < 0.001. n = 9 replicates from the differentiation of 3 individual hiPSC lines. (J) Schematic summarizing the strategy for induction of sunitinib-induced vascular dysfunction in mice. (K) Representative images of SND1 and CD31 coimmunostaining of heart sections from the mice described in J. Scale bars: 10 μm and 2 μm. (L) Representative images of SND1 and α-actinin or vimentin coimmunostaining of heart sections from mice described in J. Scale bars: 10 μm and 2 μm. (M) Quantification of the SND1+CD31+ (n = 6), SND1+ α-actinin+ (vehicle, n = 5; sunitinib, n = 6), and SND1+vimentin+ (vehicle, n = 5; sunitinib, n = 6) areas. Two-tailed Student’s t test. Data are presented as mean ± SD. ***P < 0.001. (N) Immunoblot analysis of isolated MCECs from the mice after sunitinib (Sun) (10 and 21 days) or vehicle (21 days) treatment. One-way ANOVA. Data are presented as mean ± SD. ***P < 0.001. n = 7. (O–Q) Flow cytometry analysis of SND1 expression in different cardiac macrophage subsets and cardiac monocytes, T cells, B cells, and ECs in hearts from sunitinib-treated (21 days) or vehicle-treated (21 days) mice. Two-tailed Student’s t test. Data are presented as mean ± SD. *P < 0.05. n = 3.