Jeremy Hall, Zhuoran Zhang, Supriyo Bhattacharya, Dongfang Wang, Marice Alcantara, Yong Liang, Piotr Swiderski, Stephen Forman, Larry Kwak, Nagarajan Vaidehi, Marcin Kortylewski
{"title":"用于细胞选择性和靶向降解活化 STAT3 的寡聚物-PROTAC 策略","authors":"Jeremy Hall, Zhuoran Zhang, Supriyo Bhattacharya, Dongfang Wang, Marice Alcantara, Yong Liang, Piotr Swiderski, Stephen Forman, Larry Kwak, Nagarajan Vaidehi, Marcin Kortylewski","doi":"10.1016/j.omtn.2024.102137","DOIUrl":null,"url":null,"abstract":"<p>Decoy-oligodeoxynucleotides (ODN) allow for targeting undruggable transcription factors, such as STAT3, but their limited potency and lack of delivery methods hampered translation. To overcome these challenges, we conjugated STAT3-specific decoy to thalidomide, a ligand to cereblon in E3 ubiquitin ligase complex, to generate a proteolysis-targeting chimera (STAT3D<sup>PROTAC</sup>). STAT3D<sup>PROTAC</sup> downregulated STAT3 in target cells, but not STAT1 or STAT5. Computational modeling of the STAT3D<sup>PROTAC</sup> ternary complex predicted two surface lysines K601 and K626 in STAT3 as potential ubiquitination sites. Accordingly, K601/K626 point mutations in STAT3, as well as proteasome inhibition or cereblon deletion alleviated STAT3D<sup>PROTAC</sup> effect. Next, we conjugated STAT3D<sup>PROTAC</sup> to a CpG oligonucleotide targeting Toll-like receptor 9 (TLR9) to generate myeloid/B-cell-selective C-STAT3D<sup>PROTAC</sup>. Naked C-STAT3D<sup>PROTAC</sup> was spontaneously internalized by TLR9+ myeloid cells, B-cells, human and mouse lymphoma cells but not by T cells. C-STAT3D<sup>PROTAC</sup> effectively decreased STAT3 protein levels and also STAT3-regulated target genes critical for lymphoma cell proliferation and/or survival (<em>BCL2L1, CCND2, MYC</em>). Finally, local C-STAT3D<sup>PROTAC</sup> administration to human Ly3 lymphoma-bearing mice triggered tumor regression while control C-STAT3D and C-SCR treatments had limited effects. Our results underscore feasibility of using PROTAC strategy for cell-selective, decoy oligonucleotide-based STAT3 targeting of and potentially other tumorigenic transcription factors for cancer therapy.</p>","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":6.5000,"publicationDate":"2024-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Oligo-PROTAC strategy for cell-selective and targeted degradation of activated STAT3\",\"authors\":\"Jeremy Hall, Zhuoran Zhang, Supriyo Bhattacharya, Dongfang Wang, Marice Alcantara, Yong Liang, Piotr Swiderski, Stephen Forman, Larry Kwak, Nagarajan Vaidehi, Marcin Kortylewski\",\"doi\":\"10.1016/j.omtn.2024.102137\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Decoy-oligodeoxynucleotides (ODN) allow for targeting undruggable transcription factors, such as STAT3, but their limited potency and lack of delivery methods hampered translation. To overcome these challenges, we conjugated STAT3-specific decoy to thalidomide, a ligand to cereblon in E3 ubiquitin ligase complex, to generate a proteolysis-targeting chimera (STAT3D<sup>PROTAC</sup>). STAT3D<sup>PROTAC</sup> downregulated STAT3 in target cells, but not STAT1 or STAT5. Computational modeling of the STAT3D<sup>PROTAC</sup> ternary complex predicted two surface lysines K601 and K626 in STAT3 as potential ubiquitination sites. Accordingly, K601/K626 point mutations in STAT3, as well as proteasome inhibition or cereblon deletion alleviated STAT3D<sup>PROTAC</sup> effect. Next, we conjugated STAT3D<sup>PROTAC</sup> to a CpG oligonucleotide targeting Toll-like receptor 9 (TLR9) to generate myeloid/B-cell-selective C-STAT3D<sup>PROTAC</sup>. Naked C-STAT3D<sup>PROTAC</sup> was spontaneously internalized by TLR9+ myeloid cells, B-cells, human and mouse lymphoma cells but not by T cells. C-STAT3D<sup>PROTAC</sup> effectively decreased STAT3 protein levels and also STAT3-regulated target genes critical for lymphoma cell proliferation and/or survival (<em>BCL2L1, CCND2, MYC</em>). Finally, local C-STAT3D<sup>PROTAC</sup> administration to human Ly3 lymphoma-bearing mice triggered tumor regression while control C-STAT3D and C-SCR treatments had limited effects. Our results underscore feasibility of using PROTAC strategy for cell-selective, decoy oligonucleotide-based STAT3 targeting of and potentially other tumorigenic transcription factors for cancer therapy.</p>\",\"PeriodicalId\":18821,\"journal\":{\"name\":\"Molecular Therapy. Nucleic Acids\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.5000,\"publicationDate\":\"2024-02-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular Therapy. 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Oligo-PROTAC strategy for cell-selective and targeted degradation of activated STAT3
Decoy-oligodeoxynucleotides (ODN) allow for targeting undruggable transcription factors, such as STAT3, but their limited potency and lack of delivery methods hampered translation. To overcome these challenges, we conjugated STAT3-specific decoy to thalidomide, a ligand to cereblon in E3 ubiquitin ligase complex, to generate a proteolysis-targeting chimera (STAT3DPROTAC). STAT3DPROTAC downregulated STAT3 in target cells, but not STAT1 or STAT5. Computational modeling of the STAT3DPROTAC ternary complex predicted two surface lysines K601 and K626 in STAT3 as potential ubiquitination sites. Accordingly, K601/K626 point mutations in STAT3, as well as proteasome inhibition or cereblon deletion alleviated STAT3DPROTAC effect. Next, we conjugated STAT3DPROTAC to a CpG oligonucleotide targeting Toll-like receptor 9 (TLR9) to generate myeloid/B-cell-selective C-STAT3DPROTAC. Naked C-STAT3DPROTAC was spontaneously internalized by TLR9+ myeloid cells, B-cells, human and mouse lymphoma cells but not by T cells. C-STAT3DPROTAC effectively decreased STAT3 protein levels and also STAT3-regulated target genes critical for lymphoma cell proliferation and/or survival (BCL2L1, CCND2, MYC). Finally, local C-STAT3DPROTAC administration to human Ly3 lymphoma-bearing mice triggered tumor regression while control C-STAT3D and C-SCR treatments had limited effects. Our results underscore feasibility of using PROTAC strategy for cell-selective, decoy oligonucleotide-based STAT3 targeting of and potentially other tumorigenic transcription factors for cancer therapy.
期刊介绍:
Molecular Therapy Nucleic Acids is an international, open-access journal that publishes high-quality research in nucleic-acid-based therapeutics to treat and correct genetic and acquired diseases. It is the official journal of the American Society of Gene & Cell Therapy and is built upon the success of Molecular Therapy. The journal focuses on gene- and oligonucleotide-based therapies and publishes peer-reviewed research, reviews, and commentaries. Its impact factor for 2022 is 8.8. The subject areas covered include the development of therapeutics based on nucleic acids and their derivatives, vector development for RNA-based therapeutics delivery, utilization of gene-modifying agents like Zn finger nucleases and triplex-forming oligonucleotides, pre-clinical target validation, safety and efficacy studies, and clinical trials.