Targeting MED31-driven transcription recycling in lethal prostate cancer

靶向致死性前列腺癌中 MED31 驱动的转录循环

• 批准号: 10750456
• 负责人: Qianben Wang
• 金额: $ 50.78万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750456
• 关键词: Amino Acids; Androgen Receptor; Biochemical; Biological Assay; Biological Process; Cancer Cell Growth; Cancer Patient; Castration; Cell Culture Techniques; Cells; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA; Disease; Drug Design; European; Future; Genes; Genetic Transcription; Genomics; Growth; Guide RNA; Immunohistochemistry; In Vitro; Invaded; Legal patent; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Mediator; Messenger RNA; Metastatic Neoplasm to the Liver; Modeling; Molecular; Mus; Neoplasm Metastasis; Non-Visceral; Nucleic Acids; Oncogenic; Organ; Outcome; Output; Patients; Phase; Polymerase; Process; Prognosis; Prostate; Proteins; Proteomics; RNA; RNA Polymerase II; Receptor Inhibition; Receptor Signaling; Recycling; Resistance; Role; Safety; Sampling; Specificity; Surface; System; Testing; Therapeutic; Tissue Microarray; Tissues; Toxic effect; Transcription Elongation; Transcription Initiation; Transcription Process; Treatment Efficacy; Xenograft procedure; aptamer; bone; cancer cell; castration resistant prostate cancer; clinically relevant; domain mapping; gene therapy; human tissue; improved; in vivo; insight; knock-down; lipid nanoparticle; lymph nodes; nanoparticle; nanotherapy; new therapeutic target; novel; patient derived xenograft model; preclinical development; prostate cancer cell; prostate cancer cell line; prostate cancer progression; protein expression; response; small molecule; targeted agent; targeted delivery; targeted treatment; tumorigenesis

Project Summary/Abstract Patients with lethal castration-resistant prostate cancer (CRPC) are currently treated with agents targeting androgen receptor (AR) signaling. However, AR inhibition has not dramatically improved CRPC patient survival, underscoring the need to discover novel oncogenic mechanisms in CRPC and develop new therapies targeting these mechanisms. Cancer cells are addicted to aberrant RNA polymerase II (Pol II) transcription, which includes initiation, elongation, and termination phases, as well as a recycling step critical to repeated Pol II transcription of the same gene after the initial transcription cycle. While studies have already indicated that uncontrolled transcriptional initiation and elongation have oncogenic roles, it is unknown whether other Pol II transcription processes contribute to cancer-relevant transcriptional outcomes and cancer growth. In preliminary studies, our newly developed in vitro and cell-based transcription recycling assays have found that Pol II recycling is a key yet overlooked transcription process with relevance to prostate cancer. We have found that Mediator complex subunit 31 (MED31) drives Pol II recycling in CRPC cells, enhancing mRNA output during the recycling process. Importantly, high expression of MED31 is both sufficient and necessary for prostate cancer castration-resistant growth and is associated with poor prognosis of CRPC patients. While these findings identify the oncogenic MED31 as a new therapeutic target for CRPC, transcription regulators such as MED31 are generally considered untargetable by traditional, small molecule-based drug design. We have developed a safe lipid nanoparticle (LNP) system for targeted delivery of the CRISPR/Cas13d system to efficiently and specifically knock down oncogenic transcription regulators at the mRNA level. In preliminary studies, we have demonstrated that the LNP-Cas13d system effectively and safely knocks down MED31 mRNA and decreases CRPC cell growth in vivo, establishing the proof of the concept that the therapeutic window exists for targeting MED31 in CRPC. Together, our preliminary findings support the hypothesis that MED31-governed transcription recycling is a novel oncogenic driver for CRPC progression and that an LNP-Cas13d-based RNA targeting system can counteract oncogenic transcription driven by MED31 in CRPC with safety, specificity, and efficacy. In Aim 1, we will delineate the molecular mechanism, biological function, and clinical relevance of MED31-mediated transcription recycling. In Aim 2, we will target MED31-mediated transcription recycling using a CRISPR/Cas13d-based nanoparticle system. The successful completion of these aims will significantly elucidate the critical role of Pol II recycling in lethal prostate cancer and will provide an experimental basis for future clinical trials testing the utility of an LNP- Cas13d RNA targeting system to target this novel oncogenic mechanism in CRPC patients.

项目摘要/摘要 目前，靶向药物治疗耐致命的cast割前列腺癌（CRPC）患者 雄激素受体（AR）信号传导。但是，AR抑制作用并未显着改善CRPC患者的存活， 强调需要发现CRPC中新型的致癌机制并开发针对的新疗法 这些机制。癌细胞沉迷于异常RNA聚合酶II（POL II）转录，其中包括 启动，伸长和终止阶段，以及对重复POL II转录至关重要的回收步骤 初始转录周期后同一基因的同一基因。虽然研究已经表明不受控制 转录启动和伸长具有致癌作用，尚不清楚其他POL II转录是否 过程有助于与癌症相关的转录结果和癌症的增长。在初步研究中，我们 新开发的体外和基于细胞的转录回收测定法发现，Pol II回收是关键 然而，与前列腺癌相关的转录过程被忽略了。我们发现调解人复合体 亚基31（MED31）驱动CRPC细胞中的Pol II回收，从而在回收过程中增强了mRNA输出。 重要的是，Med31的高表达对于前列腺癌cast割是足够的和必要的 生长，与CRPC患者的预后不良有关。尽管这些发现确定了致癌 MED31是CRPC的新治疗靶标，通常认为转录调节剂（例如MED31） 传统的基于小分子的药物设计不可限制。我们已经开发了一个安全的脂质纳米颗粒 （LNP）针对CRISPR/CAS13D系统有针对性交付的系统，以有效，专门敲击 mRNA水平的致癌转录调节剂。在初步研究中，我们证明了 LNP-CAS13D系统有效，安全地击倒Med31 mRNA并降低体内CRPC细胞的生长， 建立一个概念的证据，即在CRPC中靶向Med31的治疗窗口存在。一起， 我们的初步发现支持了以下假设 CRPC进展的致癌驱动器和基于LNP-Cas13d的RNA靶向系统可以抵消 由MED31在CRPC中驱动的致癌转录具有安全性，特异性和功效。在AIM 1中，我们将描绘 Med31介导的转录回收的分子机制，生物学功能和临床相关性。 在AIM 2中，我们将使用基于CRISPR/CAS13D的纳米颗粒来靶向MED31介导的转录回收 系统。这些目标的成功完成将显着阐明Pol II回收在 致命的前列腺癌，将为将来的临床试验提供实验基础，以测试LNP-的实用性 CAS13D RNA靶向系统，以瞄准CRPC患者的这种新型致癌机制。