Role of the tumor NLRP3 inflammasome in the generation of anti-PD-1 antibody immunotherapy-associated toxicities

检查点抑制剂免疫疗法在临床肿瘤学中的最新成功和扩大使用 导致多种免疫相关不良事件（irAE）的发生率增加，其中一些 这会对我们病人的生活产生持久的影响。佐剂的持续发展 免疫治疗方案和更有效的免疫治疗组合，例如 ipilimumab/nivolumab方案预期会增加这些irAE的发生率和社会影响。 尽管其患病率不断增加，但对个体irAE的潜在发病机制知之甚少 我们对这些疾病的治疗仍然很粗糙。 使用黑色素瘤的基因工程模型，我们发现抗PD-1抗体（ab） 免疫治疗通常导致肺实质和结肠板层的嗜酸性浸润 固有，与人类检查点通道相关性肺炎中观察到的病理学非常相似 和结肠炎。这项工作进一步揭示了抗PD-1 ab治疗非荷瘤小鼠， 小鼠消除了这些irAE的发展，这表明存在肿瘤内在的促炎因子。 炎症机制作为这项工作的一部分，我们最近发现了一种肿瘤内源性PD- L1：NLRP 3：HSP 70信号轴，驱动中性粒细胞在远端器官中的积聚，包括 肺和结肠对PD-1阻断的反应。其他研究表明，基因敲- 肿瘤外HSP 70表达和NLRP 3炎性体的药理学抑制 抑制中性粒细胞在这些远端组织中的积聚。基于这些发现，我们现在 假设肿瘤NLRP 3炎性体促进检查点抑制剂的发展- NLRP 3是一个很有前途的药理学靶点， 抑制这些irAE的发展。为了解决这个假设，我们已经生成了一个 转基因黑色素瘤组织特异性NLRP 3-/-模型，以检查肿瘤内源性NLRP 3在 检查点通道诱导的结肠炎和肺炎的发展。更多的研究将涉及 这种肿瘤NLRP 3途径在这些远端器官组织中的Th 17分化中的潜在作用， NLRP 3的小分子抑制剂是否是优于类固醇的上级选择 在接受抗PD-1治疗的自体黑色素瘤模型中管理结肠炎和肺炎 ab免疫疗法。我们将进一步利用肿瘤组织和DNA收集从病人接受 检查点抑制剂免疫治疗在IRB批准的临床研究中进行，以确定遗传 NLRP 3的变异可能有助于这些irAE的发展。总的来说，这项工作将提供 对irAE发展的独特机制见解，并确定合理的治疗策略 和预测性生物标志物，用于改善对接受免疫治疗的患者的管理。

The recent success and expanded use of the checkpoint inhibitor immunotherapies in clinical oncology has resulted in an increased incidence of a variety of immune-related adverse events (irAEs), some of which can have a lasting impact on the lives of our patients. The continued development of adjuvant immunotherapy protocols and more potent immunotherapy combinations such as the ipilimumab/nivolumab regimen is expected to increase the incidence and societal impact of these irAEs. Despite their growing prevalence, the underlying pathogenesis of individual irAEs is poorly understood and our therapeutic management of these conditions remains crude. Using a genetically engineered model of melanoma, we have found that anti-PD-1 antibody (ab) immunotherapy routinely leads to neutrophilic infiltration of the lung parenchyma and colon lamina propria, closely resembling the pathology noted in human checkpoint inhibitor-associated pneumonitis and colitis, respectively. This work further revealed that anti-PD-1 ab treatment of non-tumor-bearing mice eliminates the development of these irAEs, suggesting the presence of a tumor-intrinsic pro- inflammatory mechanism. As part of this work, we have recently identified a tumor-intrinsic PD- L1:NLRP3:HSP70 signaling axis that drives the accumulation of neutrophils in distant organs including the lungs and colon in response to PD-1 blockade. Additional studies have shown that genetic knock- out of tumor HSP70 expression and the pharmacological inhibition of the NLRP3 inflammasome suppresses the accumulation of neutrophils in these distant tissues. Based on these findings, we now hypothesize that the tumor NLRP3 inflammasome promotes the development of checkpoint inhibitor- associated colitis and pneumonitis and that NLRP3 represents a promising pharmacological target for suppressing the development of these irAEs. To address this hypothesis, we have generated a transgenic melanoma tissue-specific NLRP3-/- model to examine the role of tumor-intrinsic NLRP3 in the development of checkpoint inhibitor-induced colitis and pneumonitis. Additional studies will address a potential role for this tumor NLRP3 pathway in Th17 differentiation in these distant organ tissues and whether a small molecule inhibitor of NLRP3 would be a superior option over steroids for the management of colitis and pneumonitis in an autochthonous melanoma model undergoing anti-PD-1 ab immunotherapy. We will further utilize tumor tissue and DNA collected from patients undergoing checkpoint inhibitor immunotherapy on an active IRB-approved clinical study to determine if genetic variations of NLRP3 may contribute to the development of these irAEs. Overall, this work will provide unique mechanistic insight into the development of irAEs and identify rational therapeutic strategies and predictive biomarkers for improving the management of patients undergoing immunotherapy.