PDF(Pubmed)
Abstract：
NOD-Like Receptor Family Pyrin Domain Containing 3 (NLRP3) inflammasome modulation has emerged as a potential therapeutic approach targeting inflammation amplified by pyroptotic innate immune cell death. In diseases characterized by non-cell autonomous neurodegeneration including amyotrophic lateral sclerosis (ALS), the activation of several inflammasomes has been reported. Since functional redundancy can exist among inflammasome pathways, here we investigate the effects of NLRP3 inhibition on NLRP3, NLR family CARD Domain Containing 4 (NLRC4) and non-canonical pathways to understand whether NLRP3 blockade alone can mitigate pro-inflammatory cytokine release and pyroptotic cell death in contexts where single or multiple inflammasome pathways independent of NLRP3 are activated. In this study we do not limit our insights into inflammasome biology by solely relying on the THP-1 monocytic line under the LPS/nigericin-mediated NLRP3 pathway activation paradigm. We assess therapeutic potential and limitations of NLRP3 inhibition in multi-inflammasome activation contexts utilizing various human cellular systems including cell lines expressing gain of function (GoF) mutations for several inflammasomes, primary human monocytes, macrophages, healthy and Amyotrophic Lateral Sclerosis (ALS) patient induced pluripotent stem cells (iPSC)-derived microglia (iMGL) stimulated for canonical and non-canonical inflammasome pathways. We demonstrate that NLRP3 inhibition can modulate the NLRC4 and non-canonical inflammasome pathways; however, these effects differ between immortalized, human primary innate immune cells, and iMGL. We extend our investigation in more complex systems characterized by activation of multiple inflammasomes such as the SOD1G93A mouse model. Through deep immune phenotyping by single-cell mass cytometry we demonstrate that acute NLRP3 inhibition does not ameliorate spinal cord inflammation in this model. Taken together, our data suggests that NLRP3 inhibition alone may not be sufficient to address dynamic and complex neuroinflammatory pathobiological mechanisms including dysregulation of multiple inflammasome pathways in neurodegenerative disease such as ALS.
摘要：
NOD样受体家族Pyrin结构域包含3（NLRP3）炎性体调节已成为一种潜在的治疗方法，靶向通过热变性先天性免疫细胞死亡扩增的炎症。在以包括肌萎缩侧索硬化症（ALS）在内的非细胞自主性神经变性为特征的疾病中，已经报道了几种炎性体的激活。由于炎症小体途径之间可能存在功能冗余，在这里,我们研究了NLRP3抑制对NLRP3,NLR家族CARD结构域包含4(NLRC4)和非规范通路的影响,以了解在独立于NLRP3的单个或多个炎性体通路被激活的情况下,单独的NLRP3阻断是否可以减轻促炎细胞因子释放和细胞凋亡.在这项研究中，我们并不限制我们对炎症小体生物学的见解，仅依靠LPS/尼格林介导的NLRP3途径激活范式下的THP-1单核细胞系。我们评估了NLRP3抑制在多炎症体激活环境中的治疗潜力和局限性，利用各种人类细胞系统，包括表达几种炎症体功能增益（GoF）突变的细胞系，原代人单核细胞，巨噬细胞，健康和肌萎缩性侧索硬化症（ALS）患者诱导的多能干细胞（iPSC）衍生的小胶质细胞（iMGL）刺激了规范和非规范的炎性小体途径。我们证明NLRP3抑制可以调节NLRC4和非经典炎性体途径；然而，这些影响在永生化、人类初级先天免疫细胞，和iMGL。我们将研究扩展到更复杂的系统中，该系统的特征是激活多个炎性体，例如SOD1G93A小鼠模型。通过单细胞质谱术的深度免疫表型分析，我们证明了急性NLRP3抑制不能改善该模型中的脊髓炎症。一起来看,我们的数据表明,在ALS等神经退行性疾病中,仅抑制NLRP3可能不足以解决动态和复杂的神经炎症病理生物学机制,包括多个炎性体通路的失调.
