(PQ8) Biomarker identification by mass cytometry in peripheral blood of patients with renal cell carcinoma undergoing immune checkpoint therapy

项目摘要 肾癌（肾细胞癌，RCC）现在是男性中第6和第10大最常见的癌症， 女人，分别。转移性RCC（mRCC）患者的中位总生存期在2010年有所改善， 靶向治疗时代，但在14个月时仍然适度。免疫检查点抑制剂（ICI）显示出异常的 mRCC早期临床试验的前景-最终导致FDA批准nivolumab（抗PD-1）作为 2016年的第二次治疗。2018年，纳武单抗加伊匹单抗（抗细胞毒性T- 淋巴细胞相关抗原4（CTLA-4）在中度和低度恶性肿瘤中显示出显著的应答率。 风险RCC患者，最近已被批准作为一线mRCC治疗。但目前尚不清楚 为什么免疫检查点抑制剂（ICI）疗法仅对约25%的患者有效， mRCC的。人们也越来越意识到潜在的毒性，特别是免疫相关的毒性。 与检查点抑制剂相关的不良事件（irAE）。因此，迫切需要确定 1）可靠预测irAE发生和2）预测mRCC缓解的机制生物标志物 检查点抑制剂为了响应PQ 8，我们的目标是评估细胞内信号转导的变化是否 治疗前从mRCC患者中采集的外周血免疫细胞的反应可以作为 “与检查点相关的免疫相关不良事件（irAE）发生的预测生物标志物 抑制”。 我们的小组，沿着与其他人，已经表明，在外周血免疫细胞的细胞内信号反应， 与急性淋巴细胞白血病（ALL）的化疗反应、髋关节手术后的恢复、 和足月妊娠此外，最近显示，患者在i）免疫前的基线免疫状态 髋关节置换术，ii）ALL化疗，iii）转移性黑色素瘤ICI治疗 分别是恢复、复发和无进展生存期的决定因素。因此，我们的建议是建立在 假设接受ICI的mRCC患者的给药前免疫状态不同， 影响irAE的发作和临床应答。为了解决这个问题，我们将使用 测量60例mRCC患者的外周血细胞丰度和细胞内信号传导， ICI给药。在目标1中，基于我们在将CyTOF翻译成临床应用方面的经验， 样本，我们将使用新的代理标准化全血处理，使使用CyTOF“在 研究免疫抑制剂治疗前的免疫状态。在目标2中，我们将采用多元回归 识别免疫相关性以预测irAE和临床应答的算法。这些新的应用 大规模细胞计数数据集的强大统计方法将最终形成分析基础， 给药前患者特异性免疫特征、irAE和治疗疗效之间的关系。

PROJECT SUMMARY Kidney cancer (renal cell carcinoma, RCC) is now the 6th and 10th most common cancer diagnosed in men and women, respectively. Median overall survival for patients with metastatic RCC (mRCC) has improved in the targeted therapy era, but remains modest at 14 months. Immune checkpoint inhibitors (ICI) showed exceptional promise in early clinical trials for mRCC – eventually leading to FDA approval of nivolumab (anti-PD-1) as a second-line treatment in 2016. In 2018, combination therapy with nivolumab plus ipilimumab (anti-cytotoxic T- lymphocyte-associated antigen 4; CTLA-4) demonstrated significant response rates in intermediate- and poor- risk RCC patients and has recently been approved as a first-line mRCC treatment. However, it is still unclear why immune checkpoint inhibitor (ICI) therapies are effective – and remarkably so – in only ~25% of patients with mRCC. There is also increasing awareness of the potential toxicities, and specifically the immune-related adverse events (irAEs), associated with checkpoint inhibitors. Therefore, there is an urgent need to identify mechanistic biomarkers that 1) reliably predict the development of irAEs, and 2) predict the response of mRCC to checkpoint inhibitors. In response to PQ8, our goal is to evaluate whether changes in intracellular signaling responses in peripheral blood immune cells taken before treatment from patients with mRCC could serve as “predictive biomarkers for the onset of immune-related adverse events (irAEs) associated with checkpoint inhibition”. Our group, along with others, has shown that intracellular signaling responses in peripheral blood immune cells are correlated with response to chemotherapy in acute lymphoblastic leukemia (ALL), recovery from hip surgery, and full-term pregnancy. Furthermore, it was recently shown that a patient’s baseline immune state before i) surgery for hip replacement ii) chemotherapy for ALL iii) ICI treatment for metastatic melanoma were determinants of recovery, relapse and progression-free survival respectively. Our proposal is therefore built on the hypothesis that patients with mRCC receiving ICIs differ in their pre-drug immune state which will affect the onset of irAEs and clinical response. To address this, we will perform CyTOF analysis using peripheral blood measuring both cell abundance and intracellular signaling from 60 patients with mRCC before ICI administration. In Aim 1, based on our experience in streamlining the translation of CyTOF to clinical samples, we will use novel agents to standardize whole blood processing enabling the use of CyTOF “at the bedside” to study the immune state pre-treatment with ICIs. In Aim 2, we will adapt multivariate regression algorithms to identify immune correlates for predicting irAEs and clinical response. Novel applications of these powerful statistical methods to mass cytometry datasets will ultimately form the analytical groundwork to examine the relationship between patient-specific immune traits before administration of, irAEs, and treatment efficacy.