Synthetic Lethal Targeting of SETD2 in Renal Cell Carcinoma

SETD2 在肾细胞癌中的合成致死靶向

基本信息

批准号：
10607320
负责人：
Thai Huu Ho
金额：
$ 55.49万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10607320
关键词：
Affect Automobile Driving Biological Biological Assay CRISPR interference CRISPR screen Cancer Etiology Cell Line Cell Survival Cells Cessation of life ChIP-seq Chromatin Clear cell renal cell carcinoma Clinical Treatment Compensation DNA DNA Methylation Defect Disease Drug Targeting Enhancers Epigenetic Process Exhibits FDA approved Family Frequencies Gene Expression Gene Mutation Genes Genetic Genetic Epistasis Genotype Health Histone H3 Histones Homeostasis Human Immune checkpoint inhibitor Individual Injections Link Lysine Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Malignant neoplasm of prostate Mediating Methylation Modeling Mus Mutate Mutation Neoplasm Metastasis Organoids Outcome Pathway interactions Patients Phenocopy Phenotype Primary Neoplasm Proteins RNA Regulator Genes Renal Cell Carcinoma Renal carcinoma Repression Role Scheme Signal Pathway Signal Transduction Solid Neoplasm Specificity TP53 gene Tail Testing The Cancer Genome Atlas Therapeutic Tumor Tissue Tumorigenicity United States Veins Writing actionable mutation bevacizumab cell growth clinically actionable combinatorial efficacy validation epigenome genome-wide in vivo individualized medicine inhibitor loss of function loss of function mutation molecular phenotype mouse model mutant novel novel strategies novel therapeutics personalized approach pharmacologic pre-clinical response side effect small molecule inhibitor standard of care targeted treatment therapeutic target transcriptome tumor tumor growth virtual

项目摘要

ABSTRACT Clear cell renal cell carcinoma (ccRCC) accounts for ~75% of kidney cancers and is the 8th leading cause of cancer death in the United States. After completion of The Cancer Genome Atlas (TCGA) Project, clinically actionable mutations were identified in virtually every solid tumor. One major exception, however, is RCC, where the current standard of care, checkpoint inhibitor and anti-VEGF therapy, does not take into account that ~50% of RCCs have mutations in chromatin regulators. After first-line therapy, response rates are 20% and there are no FDA-approved therapies that target chromatin regulators, highlighting the need to identify how loss-of-function genotypes can be therapeutically targeted. The epigenome is profoundly disrupted in cancers including ccRCC. Aside from the near ubiquitous loss of VHL, the mutational landscape of ccRCC is dominated by loss-of-function mutations in epigenetic regulators, including SETD2, BAP1, and PBRM1. SETD2 loss has now been firmly linked to poor outcome and metastasis. The molecular phenotype of H3K36me3 deregulation in SETD2-mutant ccRCC makes this an ideal scenario to study from the angle of synthetic lethality because it induces global epigenetic changes that must be compensated for, creating unique vulnerabilities. Targeting factors that exhibit genetic epistasis with known cancer mutations to drive a synthetic lethal phenotype is a proven therapeutic approach. We performed an unbiased CRISPR/Cas9 screen to identify factors that exhibit synthetic lethality with SETD2 loss-of-function. The epigenetic factor NSD1, a writer of H3K36me1/2 acting through the H3K36 pathway, was identified. Based on these findings, we hypothesize that suppression of the H3K36 axis in the form of its epigenetic writers (NSD1 in a SETD2 loss context) drives a synthetic lethal phenotype mediated by distinct enhancer remodeling accompanied by expression defects incompatible with cell viability. Identification of NSD1, which is part of a larger family of three related proteins (NSD1/2/3) within the H3K36 signaling axis in turn, will lead to novel approaches for individualized therapeutics to target SETD2 loss-of-function, classically defined as ‘undruggable’. We will address this hypothesis with three aims. In aim 1 we functionally characterize the synthetic lethal phenotypes associated with H3K36 writers NSD1, NSD2, and NSD3 in isogenic SETD2 ccRCC cell lines. In aim 2 we demonstrate the utility of pharmacologic inhibitors of H3K36 signaling in driving synthetic lethality in SETD2-mutant cells and elucidate their biological underpinnings. Finally, in aim 3 we validate the efficacy and specificity of genetic and pharmacologic targeting of the H3K36 signaling axis to induce SETD2-mutant synthetic lethality in vivo using mouse models. Our studies will shed new light on how epigenetic regulators and the H3K36 axis specifically, drive cancer and metastasis when deregulated. This is expected to positively affect human health by generating preclinical evidence for new ways to treat SETD2-mutant ccRCC that will minimize off-target side effects, and enhance survival for patients with ccRCC, particularly those with more aggressive/metastatic disease.

抽象的透明细胞肾细胞癌（CCRCC）占肾脏癌的75％，是第8个主要原因美国的癌症死亡。癌症基因组图集（TCGA）项目完成后，临床上几乎在每个实体瘤中都鉴定出可起作的突变。但是，一个主要例外是RCC，如果目前的护理标准，检查点抑制剂和抗VEGF治疗，则没有考虑到这一点〜50％的RCC在染色质调节剂中具有突变。一线治疗后，应答率为20％，并且没有针对染色质调节剂的FDA批准的疗法，强调了需要确定如何确定如何功能丧失的基因型可以热靶向。癌症中的表观基因组在癌症中受到严重破坏包括CCRCC。除了几乎无处不在的VHL损失之外，CCRCC的突变景观是主导的通过表观遗传调节剂的功能丧失突变，包括setD2，bap1和pbrm1。 setd2损失有现在首先与不良的结果和转移有关。 H3K36me3放松管制的分子表型在setd2突变的CCRCC中，这是从合成杀伤角度研究的理想情况，因为它引起必须补偿的全球表观遗传变化，创造独特的漏洞。定位暴露了具有已知癌症突变以驱动合成致命表型的遗传性上毒的因素是一个经过验证的治疗方法。我们执行了公正的CRISPR/CAS9屏幕，以识别显示出的因素 setD2功能丧失的合成致死性。表观遗传因子NSD1，H3K36Me1/2的作者通过H3K36途径鉴定出来。基于这些发现，我们假设对 H3K36轴的表观遗传作者（setD2损失上下文中的NSD1）的形式驱动合成致死由与表达缺陷与不兼容完成的不同增强剂重塑介导的表型细胞活力。 NSD1的识别，它是三种相关蛋白质（NSD1/2/3）的较大家族的一部分 H3K36信号轴反过功能丧失，经典定义为“不可用”。我们将以三个目标解决这一假设。在目标1中我们在功能上表征了与H3K36作者NSD1，NSD2和 NSD3中的NSD3在setD2 CCRCC细胞系中。在AIM 2中，我们证明了药物抑制剂的实用性 H3K36在驱动SetD2突变细胞中驱动合成致死性的信号传导并阐明其生物学基础。最后，在AIM 3中，我们验证了遗传和药物靶向的效率和特异性使用小鼠模型在体内诱导setD2突变的合成杀伤力的H3K36信号轴的轴。我们的研究将阐明表观遗传调节剂和H3K36轴如何驱动癌症和放松管制时转移。预计这将通过产生临床前对人类健康产生积极影响治疗setD2突变ccrcc的新方法的证据，该方法将最大程度地降低目标副作用，并增强 CCRC患者，尤其是患有更具攻击性/转移性疾病的患者的生存期。