Altered mRNA splicing dependent on mutant p53 identifies novel therapeutic vulnerability in pancreatic cancer

依赖于突变体 p53 的 mRNA 剪接改变确定了胰腺癌的新治疗脆弱性

基本信息

批准号：
10399536
负责人：
Luisa Escobar Hoyos
金额：
$ 24.57万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10399536
关键词：
Acute Myelocytic Leukemia Affect Alternative Splicing Area Award Binding Biochemical Biological Biological Assay Biology Chemicals Data Dependence Development Disease Dysmyelopoietic Syndromes Electrophoretic Mobility Shift Assay Elements Environment Epidemiology Exons Faculty Familiarity Foundations Gene Expression Genetic Transcription Goals Growth Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Human Immunoprecipitation In Vitro Junior Physician KRAS2 gene KRASG12D Knowledge Laboratories Lesion Maintenance Malignant - descriptor Malignant neoplasm of pancreas Memorial Sloan-Kettering Cancer Center Mentors Messenger RNA Modeling Molecular Molecular Epidemiology of Cancer Monomeric GTP-Binding Proteins Mus Mutate Mutation Oncoproteins Pancreatic Ductal Adenocarcinoma Pancreatic Intraepithelial Neoplasia Pancreatic duct Pathogenesis Patients Pattern Poly C Population Proline Property Protein Isoforms Proteins Purines RNA RNA Processing RNA Splicing Regulation Reporter Research Research Personnel Research Training Role Scientist Signal Pathway Signal Transduction Spliceosomes Students TP53 gene Talents Testing Therapeutic Training Tumor Suppressor Proteins Universities Variant Work anticancer research base biomarker discovery biomarker validation cancer biomarkers cancer predisposition crosslink gain of function implantation in vivo in vivo Model investigator training mRNA Expression member metaplastic cell transformation molecular subtypes mutant novel therapeutics pancreas development pancreatic cancer patients pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model pancreatic neoplasm pre-clinical premalignant recruit response rho small molecule inhibitor survival outcome transcriptome sequencing transcriptomics tumor tumor growth tumorigenic

项目摘要

PROJECT SUMMARY AND ABSTRACT CANDIDATE: I am a postdoctoral research fellow co-mentored by Drs. Steve Leach and Omar Abdel-Wahab at Memorial Sloan Kettering Cancer Center (MSKCC). I have research training background in population and molecular epidemiology, cancer predisposition, cancer biomarker discovery and validation, and molecular and cellular approaches to dissect signaling pathways in disease. My current research focuses on the role of mutant p53 and aberrant alternative splicing in pancreatic ductal adenocarcinoma (PDAC). My goal as an applicant of the K99 award is to pursue answers to the research questions where we hope to identify therapeutic opportunities for pancreatic cancer patients. My long-term goal is to become an independent investigator with the focus of uncovering the transcriptomic foundations of highly aggressive PDAC tumors dictated by well-known driver and unexplored mutations and expression of alternatively spliced variants. To reach this goal, my objectives are to broaden my familiarity and expertise with experimental approaches necessary for the experimental plan outlined in the research strategy, develop further autonomy in research, and recruit and train talented students. RESEARCH: Recent studies have identified PDAC patient molecular subtypes based on gene expression profiles, where the subtype with the worst survival outcome displays enrichment of 1) mutations in TP53, and 2) altered expression of genes encoding the RNA processing machinery. The overarching goal of this proposal is to determine how mutated p53 regulates and depends on specific patterns of alternative mRNA splicing for promotion, initiation, and maintenance of PDAC. My preliminary data shows that the most commonly mutated form of oncoprotein p53, R175H, preferentially promotes splicing of poly-C sequences cassette exon mRNAs, while repressing the inclusion of exons with poly-purine sequences. The poly-C sequences result in the expression of proline-rich, SH3 binding domains in protein isoforms of a class of regulators of small-GTPases, including Ras and Rho. These observations led to my hypothesis that p53R175H alters RNA splicing of GTPase regulators that drive transformation of KRAS-mutant PanIN, to promote PDAC pathogenesis, and presents a potential mechanism of cellular transformation. In addition, we have found that small- molecule inhibitors of the core RNA splicing factors significantly reduced tumor growth and extended the survival of PDAC mice expressing p53R172H, whereas no effect was seen in the absence p53R172H. This suggests that p53R175H’s dependence on regulating gene expression through control of AS may be a viable opportunity to therapeutically intervene and discover vulnerabilities for other p53 mutations in PDAC. ENVIRONMENT: MSKCC is an ideal place for me to perform the research outlined in this proposal and obtain the necessary training and knowledge to become an independent faculty member at a major university. First, I am being mentored by Dr. Leach, the director of the David M. Rubenstein Center for Pancreatic Cancer Research and an investigator recognized for his groundbreaking work on the molecular mechanisms regarding the biology of pancreatic cancer, pancreatic neoplasia and development. In addition, I’m being co-mentored by Dr. Abdel-Wahab, a junior physician-scientist expert in mRNA splicing regulation who has done important contributions by identifying molecular mechanisms of aberrant splicing in acute myeloid leukemia and myelodysplastic syndromes. I am convinced that this is the best environment for me to transition to becoming an independent investigator and training in areas required to become an expert in the fields of RNA and PDAC biology.

项目概要和摘要候选人：我是纪念馆的博士后研究员，由 Steve Leach 和 Omar Abdel-Wahab 博士共同指导。我有人口和分子流行病学研究培训背景，斯隆凯特琳癌症中心 (MSKCC)。癌症易感性、癌症生物标志物的发现和验证以及分子和细胞分析方法我目前的研究重点是突变型 p53 和异常选择性剪接在疾病中的作用。作为 K99 奖的申请人，我的目标是寻找胰腺导管腺癌 (PDAC) 的答案。我们希望通过研究问题为胰腺癌患者找到治疗机会。是成为一名独立研究者，重点是揭示高度侵袭性的转录组学基础 PDAC 肿瘤由众所周知的驱动因素和未探索的突变以及选择性剪接变体的表达决定。为了实现这一目标，我的目标是扩大我对实验方法所需的熟悉程度和专业知识研究战略中概述的实验计划，进一步发展研究自主权，招募和培养人才学生。研究：最近的研究根据基因表达谱确定了 PDAC 患者分子亚型，其中生存结果最差的亚型表现出 1) TP53 突变的富集，以及 2) 表达的改变该提案的首要目标是确定 p53 的突变方式。调节并依赖于选择性 mRNA 剪接的特定模式来促进、启动和维持 PDAC。我的初步数据显示，最常见的癌蛋白 p53 突变形式，R175H，优先。促进多聚 C 序列盒外显子 mRNA 的剪接，同时抑制多聚嘌呤外显子的包含 Poly-C 序列导致富含脯氨酸的 SH3 结合域在蛋白质亚型中表达。小 GTP 酶的一类调节因子，包括 Ras 和 Rho 这些观察结果得出了我的假设：p53R175H。改变驱动 KRAS 突变 PanIN 转化的 GTPase 调节因子的 RNA 剪接，从而促进 PDAC 发病机制，并提出了细胞转化的潜在机制。此外，我们还发现了小- 核心RNA剪接因子的分子抑制剂显着减少肿瘤生长并延长PDAC的生存期表达 p53R172H 的小鼠，而在缺乏 p53R172H 的小鼠中没有观察到任何影响，这表明 p53R175H 的依赖性。通过控制 AS 来调节基因表达可能是治疗干预和发现的可行机会 PDAC 中其他 p53 突变的脆弱性。环境：MSKCC 是我进行本提案中概述的研究并获得成果的理想场所成为一所主要大学的独立教员所需的培训和知识首先，我正在。由 David M. Rubenstein 胰腺癌研究中心主任 Leach 博士指导研究人员因其在胰腺生物学分子机制方面的开创性工作而受到认可此外，我还得到了大三学生 Abdel-Wahab 博士的共同指导。 mRNA 剪接调控领域的医师科学家专家，通过识别分子剪接做出了重要贡献我确信这就是急性髓系白血病和骨髓增生异常综合征中异常剪接的机制。为我过渡成为一名独立调查员提供了最好的环境，并提供了所需领域的培训成为RNA和PDAC生物学领域的专家。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Aberrant RNA Splicing in Cancer.

癌症中的异常 RNA 剪接。

DOI：
10.1146/annurev-cancerbio-030617-050407
发表时间：
2019-03-01
期刊：
Annual review of cancer biology
影响因子：
0
作者：
Luisa F. Escobar;Katherine Knorr;O. Abdel
通讯作者：
O. Abdel

Keratin 17 testing in pancreatic cancer needle aspiration biopsies predicts survival.

胰腺癌针吸活检中的角蛋白 17 检测可预测生存率。

DOI：
发表时间：
2021
期刊：
影响因子：
3.4
作者：
Roa;Babu, Sruthi;Leiton, Cindy V;Wu, Maoxin;Taboada, Sofia;Akalin, Ali;Buscaglia, Jonathan;Escobar;Shroyer, Kenneth R
通讯作者：
Shroyer, Kenneth R

Keratin 17 Is a Novel Cytologic Biomarker for Urothelial Carcinoma Diagnosis.

角蛋白 17 是一种用于诊断尿路上皮癌的新型细胞学生物标志物。

DOI：
发表时间：
2021-10-13
期刊：
影响因子：
3.5
作者：
Babu, Sruthi;Kim, Nam W;Wu, Maoxin;Chan, Ina;Escobar;Shroyer, Kenneth R
通讯作者：
Shroyer, Kenneth R

Dissecting the Oncogenic Roles of Keratin 17 in the Hallmarks of Cancer.

剖析角蛋白 17 在癌症标志中的致癌作用。

DOI：
发表时间：
2022-04-01
期刊：
影响因子：
11.2
作者：
Baraks, Gabriella;Tseng, Robert;Pan, Chun;Kasliwal, Saumya;Leiton, Cindy V;Shroyer, Kenneth R;Escobar
通讯作者：
Escobar

Keratin 17 identifies the most lethal molecular subtype of pancreatic cancer.

角蛋白 17 确定了胰腺癌最致命的分子亚型。

DOI：
发表时间：
2019
期刊：
影响因子：
4.6
作者：
Roa;Leiton, Cindy V;Babu, Sruthi;Pan, Chun;Vanner, Elizabeth A;Akalin, Ali;Bandovic, Jela;Moffitt, Richard A;Shroyer, Kenneth R;Escobar
通讯作者：
Escobar