Complement therapeutics meets nanomedicine: overcoming human complement activation and leukocyte uptake of nanomedicines with soluble domains of CD55
Complement therapeutics meets nanomedicine: overcoming human complement activation and leukocyte uptake of nanomedicines with soluble domains of CD55
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DOI：
10.1016/j.jconrel.2019.04.009
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发表时间：
2019-05-28
影响因子：
10.8
通讯作者：
Simberg, Dmitri
中科院分区：
文献类型：
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作者：
Gifford, Geoffrey;Vu, Vivian P.;Simberg, Dmitri
补体活化在静脉给药纳米药物的药代动力学和性能中起着重要作用。对于低补体激活的纳米表面的工程研究已经取得了很大的进展，但由于补体因子的混杂性和途径的冗余性，这仍然是一个重大的挑战。细胞膜锚定衰变加速因子（DAF，又名CD55）是一种有效的膜结合补体调节剂，通过加速经典和替代C3转化酶的自发衰变来抑制它们。在这里，我们测试了人类CD55的各种短一致重复序列（SCRs，“sushi”结构域）对人类血清和血浆中纳米颗粒介导的补体激活的影响。结构建模表明，SCR-2、SCR-3和SCR-4对于结合替代途径C3bBb转化酶至关重要，而SCR-1则是可缺性的。多种结构域在大肠杆菌中表达，并通过亲和柱纯化。将SCRs添加到不同健康受试者的lepirudin血浆或血清中，以监测纳米颗粒介导的补体活化和C3活化。使用超顺磁氧化铁纳米蠕虫（SPIO NWs），我们发现SCR-2-3-4是最有效的抑制剂（IC50类似于0.24 μ M），其次是SCR-1-2-3-4 (IC50类似于0.6 μ M)，而较短的结构域（SCR-3, SCR-2-3, SCR 34）无效。SCR-2-3-4对C5a的产生也有抑制作用（血清中IC50与0.16 μ M相似）。除了SPIO NWs外，SCR-2-3-4还能有效抑制临床批准的纳米颗粒（Feraheme、LipoDox和Onivyde）对C3的活化和C5a的产生。SCR-2-3-4抑制凝集素和纳米颗粒的替代途径激活。当添加到健康供者的lepirudi抗凝血中时，它显著降低了中性粒细胞和单核细胞对SPIO NWs的摄取。这些结果表明，膜结合补体抑制剂的可溶性结构域是防止纳米药物介导的人类补体活化的潜在候选者。
Complement activation plays an important role in pharmacokinetic and performance of intravenously administered nanomedicines. Significant efforts have been directed toward engineering of nanosurfaces with low complement activation, but due to promiscuity of complement factors and redundancy of pathways, it is still a major challenge. Cell membrane-anchored Decay Accelerating Factor (DAF, a.k.a. CD55) is an efficient membrane bound complement regulator that inhibits both classical and alternative C3 convertases by accelerating their spontaneous decay. Here we tested the effect of various short consensus repeats (SCRs, "sushi" domains) of human CD55 on nanoparticle-mediated complement activation in human sera and plasma. Structural modeling suggested that SCR-2, SCR-3 and SCR-4 are critical for binding to the alternative pathway C3bBb convertase, whereas SCR-1 is dispensable. Various domains were expressed in E.coli and purified by an affinity column. SCRs were added to lepirudin plasma or sera from different healthy subjects, to monitor nanoparticle-mediated complement activation as well as C3 opsonization. Using superparamagnetic iron oxide nanoworms (SPIO NWs), we found that SCR-2-3-4 was the most effective inhibitor (IC50 similar to 0.24 mu M for C3 opsonization in sera), followed by SCR-1-2-3-4 (IC50 similar to 0.6 mu M), whereas shorter domains (SCR-3, SCR-2-3, SCR 3 4) were ineffective. SCR-2-3-4 also inhibited C5a generation (IC50 similar to 0.16 mu M in sera). In addition to SPIO NWs, SCR-2-3-4 effectively inhibited C3 opsonisation and C5a production by clinically approved nanoparticles (Feraheme, LipoDox and Onivyde). SCR-2-3-4 inhibited both lectin and alternative pathway activation by nanoparticles. When added to lepirudi-nanticoagulated blood from healthy donors, it significantly reduced the uptake of SPIO NWs by neutrophils and monocytes. These results suggest that soluble domains of membrane-bound complement inhibitors are potential candidates for preventing nanomedicine-mediated complement activation in human subjects.