Targeting Small Nucleolar RNA Augments Immunotherapeutic Efficacy

项目摘要 尽管肥胖被确定为多种癌症类型发展的风险因素，包括 在乳腺癌的发病机制中，仍有明显不足的遗传学证据表明肥胖与其发病有关。 和乳腺癌发展的进展。而免疫治疗方法，如PD-1/PD-L1 封锁治疗乳腺癌前景看好，肥胖导致抗肿瘤功能实质受损 豁免权。开发有效的肥胖症免疫治疗策略面临的一个重大挑战- 与乳腺癌相关的是驱动肥胖相关的骨性靶基因的特征 免疫抵抗力。非编码RNA作为重要的生物分子的出现为 为开发下一代治疗策略奠定了基础。我们的初步数据表明 一种小核仁RNA(SnoRNA)SNORD46在乳腺癌中上调，并与 细胞毒性白细胞的渗入。我们还指出，肥胖与一种染色质变体 SNORD46。我们收集了遗传证据，表明SNORD46的表达会导致肥胖和 免疫治疗耐药的乳腺肿瘤。因此，这项建议的目标是 全面描述SNORD46是肥胖相关乳腺癌的驱动因素，并将其与 靶向snoRNAs可有效促进CD8T细胞和NK细胞的扩增和活化 并在免疫治疗中使肥胖相关的乳腺癌变得敏感。 我们将解决我们的核心假设，即snoRNAs作为促进肥胖和 肥胖相关乳腺癌的启动和免疫抵抗，在体内可以通过抗肥胖药物减弱 正义寡核苷酸为基础的靶向治疗。在具体目标1中，我们将定义 SNORD46抑制依赖IL-15的脂肪细胞非经典途径，以及IL-15依赖的 CD8T细胞和NK细胞的刺激性检查点。在具体目标2中，我们将确定功能重要性 SnoRNAs在高脂饮食诱导的肥胖小鼠肥胖相关肿瘤发生和免疫抵抗中的作用 和Snord46 MUT/MUT敲击肥胖小鼠。 这项拟议的研究将为确定脂肪细胞表达的关键作用提供初步的遗传学证据。 SnoRNAs在促进肥胖和肥胖相关乳腺癌中的作用。从机制上讲，我们将阐明 SNORD46与白细胞相互作用导致肥胖相关乳腺癌的分子机制 免疫治疗的抵抗力。在临床上，拟议的研究将描绘使用锁定的靶向snoRNA 核酸(LNAs)在体内能有效促进CD8 T细胞和NK细胞的增殖/激活， 使肥胖相关乳腺癌对免疫检查点阻滞剂敏感。

Project Summary Despite the establishment of obesity as a risk factor in the development of a multitude of cancer types including breast cancer, there is still a significant insufficiency of genetic evidence for implicating obesity in the initiation and progression of breast cancer development. While immunotherapeutic approaches such as the PD-1/PD-L1 blockade have shown promise in treating breast cancer, obesity causes substantial impairment of anti-tumor immunity. A significant challenge in the development of effective immunotherapeutic strategies for obesity- associated breast cancer lies in the characterization of the bone fide target genes that drive obesity-associated immune resistance. The emergence of noncoding RNAs as important biological molecules has provided the foundation for the development of next-generation treatment strategies. Our preliminary data demonstrated that a small nucleolar RNA (snoRNA), SNORD46, is upregulated in breast cancer and negatively correlated with the infiltration of cytotoxic leukocytes. We also indicated that obesity is associated with a chromatin variant of SNORD46. We collected genetic evidence indicating that the expression of SNORD46 leads to obesity and mammary gland tumors which are resistant to immunotherapy. Therefore, the goal of this proposal is to comprehensively characterize and implicate SNORD46 as a driver of obesity-associated breast cancer and demonstrate that targeting snoRNAs effectively improves the expansion and activation of CD8+ T-cells and NK cells and sensitizes obesity-associate breast cancer in immunotherapy. We will address our central hypothesis that snoRNAs act as essential gene targets that promote obesity and obesity-associated breast cancer initiation and immune resistance, which could be attenuated in vivo by an anti- sense oligonucleotide-based targeted therapy. In Specific Aim 1, we will define the molecular mechanism of SNORD46 inhibited, IL-15-dependent non-classic pathway in adipocytes, and IL-15-dependent expression of stimulatory checkpoints in CD8+ T cell and NK cells. In Specific Aim 2, we will ascertain the functional importance of snoRNAs in obesity-associated tumorigenesis and immune resistance using high-fat diet-induced obese mice and Snord46 mut/mut knockin obese mice. The proposed study will provide initial genetic evidence for defining the crucial role of adipocyte-expressed snoRNAs in promoting obesity and obesity-associated breast cancer. Mechanistically, we will elucidate the molecular mechanisms of the SNORD46-leukocyte interactions that drive obesity-associated breast cancer resistance to immunotherapy. Clinically, the proposed studies will delineate that targeting snoRNAs using locked nucleic acids (LNAs) effectively improves the proliferation/activation of CD8+ T-cells and NK cells in vivo, sensitizing obesity-associated breast cancer to immune checkpoint blockers.