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

检查点抑制免疫疗法在临床肿瘤学中的近期成功和推广应用 导致各种免疫相关不良事件(IrAEs)的发生率增加，其中一些 这可能会对我们患者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，如 预计ipilimumab/nivolumab方案将增加这些irAEs的发病率和社会影响。 尽管其发病率越来越高，但对个体irAEs的潜在发病机制知之甚少。 我们对这些疾病的治疗方法仍然很粗糙。 利用黑色素瘤的基因工程模型，我们发现抗PD-1抗体(Ab) 免疫治疗通常会导致肺实质和结肠板的中性粒细胞浸润。 专有，与人类检查点抑制物相关性肺炎的病理非常相似 和结肠炎。这项工作进一步揭示了抗PD-1ab治疗非荷瘤 小鼠消除了这些irAEs的发展，这表明存在一种肿瘤固有的促癌基因。 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：NLRP3：HSP70信号轴，驱动中性粒细胞在远处器官中聚集，包括 肺和结肠对PD-1阻断的反应。更多的研究表明，基因敲击- 肿瘤外HSP70表达与NLRP3炎症体的药理抑制 抑制中性粒细胞在这些远处组织中的聚集。基于这些发现，我们现在 假设肿瘤NLRP3炎症体促进检查点抑制物的发展- NLRP3是治疗结肠炎和肺炎的有效药物靶点。 抑制这些irAEs的发展。为了解决这一假设，我们生成了一个 转基因黑色素瘤组织特异性NLRP3-/-模型研究肿瘤内源性NLRP3在 检查点抑制剂诱导的结肠炎和肺炎的研究进展。更多的研究将解决 该肿瘤NLRP3通路在这些远处器官组织和 NLRP3小分子抑制剂是否比类固醇更适合治疗 抗PD-1治疗原发黑色素瘤模型中结肠炎和肺炎的治疗 AB免疫疗法。我们将进一步利用从患者身上收集的肿瘤组织和DNA 检查点抑制免疫疗法在IRB批准的一项积极临床研究中的应用，以确定基因 NLRP3基因变异可能与这些irAEs的发生有关。总体而言，这项工作将提供 对irAEs发展的独特机制洞察和确定合理的治疗策略 和预测性生物标志物，用于改善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.