文献类型：期刊文献

英文题名：Complement-coupled erythrocyte platform for rapid enrichment of candidate NLRP3 modulators

作者：Shi, Lijia[1];Liu, Wenjing[1];Xie, Xueman[1];Zhang, Yuanxing[1];Liu, Qin[1];Chen, Shouwen[1]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2026

卷号：829

外文期刊名：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001806393800001)】；

基金：The authors thank the staff at the Blood Donation Center of Xijing Hospital for their assistance in blood sample collection. The authors thank Prof. Jian Li (School of Pharmacy, East China University of Science and Technology) for generously providing the small-molecule compound library. The presented work was supported by Science and Technology Commission of Shanghai Municipality (24HC2820500), the National Key Research and Development Program of China (2022YFC3400103), and the ECUST-OPM Open Fund (20220701).

语种：英文

外文关键词：Complement-mediated hemolysis; High-throughput drug discovery; NLRP3; Phenotypic screening; Spectosis

摘要：Despite the validation of NLRP3 as a therapeutic target for inflammatory disorders, conventional cell-based screens for NLRP3 modulators remain resource-intensive and technically demanding. Here, we report a phenotypic screening platform based on spectosis, a complement-driven erythrocyte death pathway mechanistically coupled to NLRP3 signaling. By monitoring complement-mediated hemolysis, this system enables highthroughput phenotypic enrichment of compounds for subsequent evaluation. A diverse library of 1000 small molecules was screened using a permissive primary threshold of 20% inhibition to ensure comprehensive coverage, yielding 365 initial actives. These underwent three rounds of confirmatory screening under identical conditions, yielding 11 validated hits that consistently passed all rounds. Importantly, counter-screening against terminal complement component C9 deposition showed that ten of these eleven compounds did not reduce C9 deposition, indicating that the majority of hits do not act via direct complement suppression. Translational validation in THP-1 macrophages identified five candidates that effectively attenuated both IL-1(3 maturation and LDH release, consistent with NLRP3 pathway modulation. This erythrocyte-based platform thus enables rapid, cost-effective enrichment of candidate chemotypes with potential NLRP3-related activity from large chemical libraries while circumventing the throughput limitations of conventional macrophage-based assays.