Targeting Small Nucleolar RNA Augments Immunotherapeutic Efficacy

项目摘要 尽管肥胖是多种癌症的危险因素， 虽然肥胖与乳腺癌的发病有关，但遗传学证据仍显不足， 和乳腺癌的发展。虽然免疫学方法如PD-1/PD-L1 阻断已显示出治疗乳腺癌的希望，肥胖导致抗肿瘤的实质性损害， 免疫力开发有效的肥胖免疫策略的一个重大挑战- 相关乳腺癌的特征在于驱动肥胖相关乳腺癌的真正靶基因的特征。 免疫抵抗非编码RNA作为重要的生物分子的出现， 为下一代治疗策略的发展奠定基础。我们的初步数据表明， 一种小核仁RNA（snoRNA），SNORD 46，在乳腺癌中表达上调，并与乳腺癌的发生呈负相关。 细胞毒性白细胞浸润。我们还指出，肥胖与一种染色质变异有关， SNORD 46.我们收集的遗传证据表明，SNORD 46的表达导致肥胖， 对免疫疗法有抵抗力的乳腺肿瘤。因此，本提案的目标是 全面表征并暗示SNORD 46是肥胖相关乳腺癌的驱动因素， 证明靶向snoRNA有效地改善了CD 8 + T细胞和NK细胞的扩增和活化。 细胞，并在免疫治疗中使肥胖相关的乳腺癌敏感。 我们将讨论我们的中心假设，即snoRNA作为促进肥胖的重要基因靶点， 肥胖相关的乳腺癌的发生和免疫抵抗，这可以在体内通过抗 基于正义寡核苷酸的靶向治疗。在具体目标1中，我们将定义 SNORD 46抑制脂肪细胞中IL-15依赖性非经典途径， CD 8 + T细胞和NK细胞中刺激性检查点。在具体目标2中，我们将确定功能重要性 使用高脂饮食诱导的肥胖小鼠研究snoRNA在肥胖相关肿瘤发生和免疫抵抗中的作用 和Snord 46 mut/mut敲入肥胖小鼠。 这项研究将提供初步的遗传学证据，以确定脂肪细胞表达的 snoRNA在促进肥胖和肥胖相关乳腺癌中的作用机械地，我们将阐明 SNORD 46-白细胞相互作用驱动肥胖相关乳腺癌的分子机制 对免疫疗法的抵抗。在临床上，所提出的研究将描述使用锁定的靶向snoRNA， 核酸（LNA）有效地改善体内CD 8 + T细胞和NK细胞的增殖/活化， 使肥胖相关的乳腺癌对免疫检查点阻断剂敏感。

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.