Generation and characterization of humanized affinity-matured EGFL6 antibodies for ovarian cancer therapy.
Generation and characterization of humanized affinity-matured EGFL6 antibodies for ovarian cancer therapy.
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DOI：
10.1016/j.ygyno.2023.02.004
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发表时间：
2023-02
影响因子：
4.7
通讯作者：
Huijuan Tang;A. Fayomi;Shoumei Bai;Navneet Gupta;S. Cascio;Dongli Yang;R. Buckanovich
中科院分区：
文献类型：
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作者：
Huijuan Tang;A. Fayomi;Shoumei Bai;Navneet Gupta;S. Cascio;Dongli Yang;R. Buckanovich
目的表皮生长因子（EGF）样结构域多聚体6（EGFL 6）在高级别浆液性卵巢癌中高表达，并促进内皮细胞增殖/血管生成和癌细胞增殖/转移。因此，它被认为是一种治疗靶点。EGFL 6作为一种分泌性因子，是抗体治疗的候选者。本研究的目的是创建和验证人源化亲和力成熟的EGFL 6中和抗体的临床development.MethodsA选择的鼠EGFL 6抗体进行人源化，使用CDR移植创建26个变体人源化抗体。这些被筛选并且先导候选物是亲和力成熟的。7人源化亲和力成熟的EGFL 6抗体筛选其阻断EGFL 6对癌细胞的活性在体外的能力，其中两个被选中，并测试其治疗activityinvivo.ResultsHumanized亲和力成熟的抗体表现出高亲和力EGFL 6（150 pM至2.67 nM）。我们发现几种人源化亲和力成熟的EGFL 6抗体特异性结合重组和天然人EGFL 6。两种先导抗体能够在体外抑制EGFL 6介导的（i）癌细胞迁移，（ii）增殖，和（iii）癌细胞中ERK磷酸化的增加。两种先导抗体均限制表达EGFL 6的卵巢癌患者来源的异种移植物的生长。治疗的人肿瘤异种移植物的分析表明，抗EGFL 6治疗抑制血管生成，抑制肿瘤细胞增殖，并促进肿瘤细胞apoptosis.ConclusionsOur研究证实这些人源化亲和力成熟的抗体，以中和EGFL 6和作为一种治疗，以限制癌症的生长能力。这项工作支持这些抗体的开发，用于首次人体临床试验。
ObjectivesEpidermal growth factor EGF-like domain multiple-6 (EGFL6) is highly expressed in high grade serous ovarian cancer and promotes both endothelial cell proliferation/angiogenesis and cancer cell proliferation/metastasis. As such it has been implicated as a therapeutic target. As a secreted factor, EGFL6 is a candidate for antibody therapy. The objectives of this study were to create and validate humanized affinity-matured EGFL6 neutralizing antibodies for clinical development.MethodsA selected murine EGFL6 antibody was humanized using CDR grafting to create 26 variant humanized antibodies. These were screened and the lead candidate was affinity matured. Seven humanized affinity-matured EGFL6 antibodies were screened for their ability to block EGFL6 activity on cancer cellsin vitro, two of which were selected and tested their therapeutic activityin vivo.ResultsHumanized affinity matured antibodies demonstrated high affinity for EGFL6 (150 pM to 2.67 nM). We found that several humanized affinity-matured EGFL6 antibodies specifically bound to recombinant, and native human EGFL6. Two lead antibodies were able to inhibit EGFL6-mediated (i) cancer cell migration, (ii) proliferation, and (iii) increase in ERK phosphorylation in cancer cellsin vitro. Both lead antibodies restricted growth of an EGFL6 expressing ovarian cancer patient derived xenograft. Analysis of treated human tumor xenografts indicated that anti-EGFL6 therapy suppressed angiogenesis, inhibited tumor cell proliferation, and promoted tumor cell apoptosis.ConclusionsOur studies confirm the ability of these humanized affinity-matured antibodies to neutralize EGFL6 and acting as a therapeutic to restrict cancer growth. This work supports the development of these antibody for first-in-human clinical trials.