Targeting RAGE in tumor and TME to oppose inflammation and drug resistance in obesity associated ER+ breast cancer

项目摘要/摘要 肥胖与绝经后雌激素受体阳性(ER+)乳腺癌(BC)增加有关 风险和所有BC亚型的死亡率增加2-4倍。治疗耐药性增加的机制 肥胖症的致命性BC转移仍不清楚。在这里，我们研究肥胖的炎症状态是如何 驱动ER+BC。我们研究了绝经后雌激素，雌酮(E1)，它在肥胖症中高出3倍，是如何与 与核因子κB和晚期糖基化终产物受体(RAGE)共同作用，上调转移。 我们的数据表明BC细胞：脂肪细胞接触激活了NFκB和E1：ERα，从而诱导了促炎反应 两种细胞类型中的细胞因子基因，刺激更大的癌症干细胞扩张，以及ER+BC的快速增长和 转移。初步数据显示，核因子κB和E1：ER共刺激编码RAGE配体的基因(S100A8/A9)。 RAGE是其他类型细胞(包括免疫和内皮细胞)中的主要核因子κB激活剂，但人们知之甚少 在BC中RAGE/NFκB信号转导的表达。我们发现RAGE在肿瘤细胞和宿主微环境中都有作用。 (间质和脂肪)上调细胞因子，招募髓系来源的抑制细胞(MDSC)促进 转移。在肥胖的小鼠中，E1促进了BC的生长，部分是通过刺激免疫逃避。新数据显示 具有抗雌激素耐药性的ER+BC株，包括那些携带esr1突变的株，显示出更高的NFκB活性和 愤怒程度。此外，研究中的RAGE抑制剂TTP488与ER阻滞剂Fulvestrant， 在多个耐药系中阻止抗雌激素耐药的ER+BC细胞的生长。在这里，我们测试e1结合的ER， 核因子κB和RAGE在ER+BC、脂肪细胞和免疫细胞中相互作用，驱动内分泌治疗的基因程序 抵抗，如果愤怒抑制剂逆转了这一点。 我们假设E1：ER和NFκB协同作用诱导RAGE，RAGE激活ER+BC中的NFκB 肿瘤细胞和瘤周脂肪驱动肿瘤促炎、促转移基因表达程序 进展与获得性耐药。目标1将识别肿瘤细胞内在的前馈机制 在富含E1的乳腺癌环境中介导RAGE/NFκB的激活，测试E1和NFκB是否诱导RAGE 在ER+BC中激活RAC1和TLR4，驱动前馈致癌的NFκB的激活。目标2将测试愤怒是否 通过E1/ER和核因子κB在乳腺癌细胞中介导促肿瘤、促炎靶基因的激活。我们 将识别E1/ER和NFκB周期和转录本，并测试这些是否需要RAGE。的关联性 RAGE依赖的ER/κB共靶基因在肥胖中的激活将通过比较ScRNAseq在 来自肥胖和瘦捐赠者的人类ER+乳腺癌。目标3将确定肿瘤细胞的外在机制 肥胖宿主瘤周脂肪促进ER+获得性抗雌激素抵抗和免疫逃避 癌症。AIM 4将在有机物和PDX模型中测试RAGE抑制剂是否能恢复内分泌治疗反应 来源于ER+乳腺癌。这项工作可以确定治疗获得性内分泌的新方法。 转移性ER+BC的耐药性，特别是在肥胖患者。

PROJECT SUMMARY/ABSTRACT Obesity is associated with increased postmenopausal estrogen receptor-positive (ER+) breast cancer (BC) risk and a 2-4 fold increase in mortality from all BC subtypes. Mechanisms of increased resistance to therapy and ensuing fatal BC metastasis in obesity remain unclear. Here, we study how the inflammatory state of obesity drives ER+ BC. We study how postmenopausal estrogen, estrone (E1), which is 3-fold higher in obesity, co- operates with NFκB and the Receptor for Advanced Glycation End-products (RAGE) to upregulate metastasis. Our data indicate that BC cell:adipocyte contact activates NFκB and E1:ERα to induce pro-inflammatory cytokine genes in both cell types, stimulating greater cancer stem cell expansion, and rapid ER+ BC growth and metastasis. Preliminary data show NFκB and E1:ER co-stimulate genes encoding RAGE-ligands (S100A8/A9). RAGE is a major NFκB activator in other cell types (including immune and endothelial cells), but little is known of RAGE/NFκB signaling in BC. We showed that RAGE acts in both tumor cells and the host microenvironment (stroma and fat) to upregulate cytokines that recruit myeloid-derived suppressor cells (MDSC) to promote metastasis. In obese mice, E1 increases BC growth, in part by stimulating immune evasion. New data show antiestrogen-resistant ER+BC lines, including those bearing ESR1 mutations, show increased NFκB activity and RAGE levels. Moreover, the investigational RAGE inhibitor, TTP488, cooperates with the ER-blocker, fulvestrant, to arrest antiestrogen-resistant ER+ BC cell growth in multiple resistant lines. Here, we test if E1-bound ER, NFκB, and RAGE interact in ER+ BC, adipocytes and immune cells to drive gene programs of endocrine therapy resistance and if RAGE inhibitors reverse this. We hypothesize that E1:ER and NFκB cooperate to induce RAGE, and RAGE activates NFκB in ER+ BC tumor cells and peritumoral fat to drive pro-inflammatory, pro-metastatic gene expression programs of tumor progression and acquired drug resistance. Aim 1 will identify tumor cell-intrinsic feed-forward mechanisms mediating RAGE/ NFκB activation in an E1-rich breast cancer environment, testing if E1 and NFκB induce RAGE to activate Rac1, and TLR4 driving feed-forward oncogenic NFκB activation in ER+ BC. Aim 2 will test if RAGE mediates pro-oncogenic, pro-inflammatory target gene activation by E1/ER and NFκB in breast cancer cells. We will identify E1/ER and NFκB cistromes and transcriptomes and test if these require RAGE. The relevance of RAGE-dependent ER/κB co-target genes activation in obesity will be validated by comparing ScRNAseq in human ER+ breast cancers from obese and lean donors. Aim 3 will identify tumor cell-extrinsic mechanisms whereby peritumoral fat in obese hosts promotes acquired antiestrogen resistance and immune evasion in ER+ cancers. Aim 4 will test if RAGE inhibitors restore endocrine therapy responses in organoid and PDX models derived from ER+ breast cancers. This work could identify new approaches to treating acquired endocrine resistance in metastatic ER+BC, particularly in obese patients.