新型选择性抗Notch1中和抗体阻断Notch1可通过促进发炎的TME提高黑色素瘤的免疫治疗效果。

IF 11.4 1区 医学 Q1 ONCOLOGY Journal of Experimental & Clinical Cancer Research Pub Date : 2024-11-04 DOI:10.1186/s13046-024-03214-5
Juliano Tiburcio de Freitas, Varsha Thakur, Kathryn M LaPorte, Vijay S Thakur, Brian Flores, Valentina Caicedo, Chioma G E Ajaegbu, Giuseppe Ingrasci, Zoe M Lipman, Keman Zhang, Hong Qiu, Thomas R Malek, Barbara Bedogni
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引用次数: 0

摘要

背景:免疫检查点抑制剂(ICI免疫检查点抑制剂(ICI)大大延长了转移性黑色素瘤患者的寿命。然而,约有一半的患者仍对这些治疗产生抗药性。我们之前已经证明,Notch1 有助于黑色素瘤的非炎症 TME,从而降低对 ICI 的反应。在此,我们探讨了我们生产的新型抗Notch1中和抗体在黑色素瘤模型中单独或与免疫检查点抑制剂联合使用的治疗效果:方法:抗 Notch1 被设计为干扰配体结合。用包含人Notch1的EGF样重复序列11-15的多肽免疫小鼠,这是配体结合和Notch1激活所需的最小区域。对阳性克隆进行扩增并测试其中和能力。抗Notch1-NIC用于确定抗Notch1是否能减少Notch1的裂解;而抗SNAP23和BCAT2则分别作为Notch1和Notch2的下游靶点。研究使用了 K457 人类黑色素瘤细胞以及 YUMM2.1 和 1.7 合成小鼠黑色素瘤细胞。通过胰蓝染色确定抗Notch1处理后的细胞死亡情况,并与γ-分泌酶抑制剂DBZ的效果进行比较。10 mg/kg anti-Notch1用于YUMM2.1和1.7细胞的体内肿瘤生长。使用针对主要免疫细胞群的抗体对肿瘤进行测量和流式细胞术处理:结果:抗 Notch1 可选择性抑制 Notch1,但不能抑制 Notch2;在体外可导致黑色素瘤细胞大量死亡,但不影响正常黑色素细胞。在体内,它能延缓肿瘤生长,且无明显的胃肠道毒性症状;重要的是,它能通过增加细胞毒性 CD8+ T 细胞而减少耐受性 Tregs 和 MDSCs 来促进炎性 TME,从而增强抗 PD-1 的疗效:结论:抗Notch1能安全地发挥抗黑色素瘤作用,并提高免疫检查点抑制剂的疗效。结论:抗Notch1能安全地发挥抗黑色素瘤的作用,并能提高免疫检查点抑制剂的疗效,因此,抗Notch1是黑色素瘤免疫疗法的新成员。
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Notch1 blockade by a novel, selective anti-Notch1 neutralizing antibody improves immunotherapy efficacy in melanoma by promoting an inflamed TME.

Background: Immune checkpoint inhibitors (ICI) have dramatically improved the life expectancy of patients with metastatic melanoma. However, about half of the patient population still present resistance to these treatments. We have previously shown Notch1 contributes to a non-inflamed TME in melanoma that reduces the response to ICI. Here, we addressed the therapeutic effects of a novel anti-Notch1 neutralizing antibody we produced, alone and in combination with immune checkpoint inhibition in melanoma models.

Methods: Anti-Notch1 was designed to interfere with ligand binding. Mice were immunized with a peptide encompassing EGF-like repeats 11-15 of human Notch1, the minimal required region that allows ligand binding and Notch1 activation. Positive clones were expanded and tested for neutralizing capabilities. Anti-Notch1-NIC was used to determine whether anti-Notch1 was able to reduce Notch1 cleavage; while anti-SNAP23 and BCAT2 were used as downstream Notch1 and Notch2 targets, respectively. K457 human melanoma cells and the YUMM2.1 and 1.7 syngeneic mouse melanoma cells were used. Cell death after anti-Notch1 treatment was determined by trypan blue staining and compared to the effects of the gamma-secretase inhibitor DBZ. 10 mg/kg anti-Notch1 was used for in vivo tumor growth of YUMM2.1 and 1.7 cells. Tumors were measured and processed for flow cytometry using antibodies against major immune cell populations.

Results: Anti-Notch1 selectively inhibited Notch1 but not Notch2; caused significant melanoma cell death in vitro but did not affect normal melanocytes. In vivo, it delayed tumor growth without evident signs of gastro-intestinal toxicities; and importantly promoted an inflamed TME by increasing the cytotoxic CD8+ T cells while reducing the tolerogenic Tregs and MDSCs, resulting in enhanced efficacy of anti-PD-1.

Conclusions: Anti-Notch1 safely exerts anti-melanoma effects and improves immune checkpoint inhibitor efficacy. Thus, anti-Notch1 could represent a novel addition to the immunotherapy repertoire for melanoma.

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来源期刊
CiteScore
18.20
自引率
1.80%
发文量
333
审稿时长
1 months
期刊介绍: The Journal of Experimental & Clinical Cancer Research is an esteemed peer-reviewed publication that focuses on cancer research, encompassing everything from fundamental discoveries to practical applications. We welcome submissions that showcase groundbreaking advancements in the field of cancer research, especially those that bridge the gap between laboratory findings and clinical implementation. Our goal is to foster a deeper understanding of cancer, improve prevention and detection strategies, facilitate accurate diagnosis, and enhance treatment options. We are particularly interested in manuscripts that shed light on the mechanisms behind the development and progression of cancer, including metastasis. Additionally, we encourage submissions that explore molecular alterations or biomarkers that can help predict the efficacy of different treatments or identify drug resistance. Translational research related to targeted therapies, personalized medicine, tumor immunotherapy, and innovative approaches applicable to clinical investigations are also of great interest to us. We provide a platform for the dissemination of large-scale molecular characterizations of human tumors and encourage researchers to share their insights, discoveries, and methodologies with the wider scientific community. By publishing high-quality research articles, reviews, and commentaries, the Journal of Experimental & Clinical Cancer Research strives to contribute to the continuous improvement of cancer care and make a meaningful impact on patients' lives.
期刊最新文献
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