Targeting CD5 chimeric antigen receptor-engineered natural killer cells against T-cell malignancies.

IF 9.4 1区医学 Q1 HEMATOLOGY

Experimental Hematology & Oncology Pub Date : 2024-10-26 DOI:10.1186/s40164-024-00577-5

Yingling Zu, Quan Ren, Jishuai Zhang, Hongchang Su, Qiumei Lu, Yongping Song, Jian Zhou

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引用次数: 0

Abstract

Background: Chimeric antigen receptor engineered T cells (CAR-T) have demonstrated promising clinical efficacy in B-cell malignancies, and the approach has been extended to T-cell malignancies. However, the use of allogeneic T cells in CAR therapy poses a challenge due to the risk of graft-versus-host disease. Recently, natural killer (NK) cells have exhibited "off‑the‑shelf" availability. The nanobody-based CAR structures have attracted much attention for their therapeutic potential owing to the advantages of nanobody, including small size, optimal stability, high affinity and manufacturing feasibility. CD5, a common surface marker of malignant T cells, has three scavenger receptor cysteine-rich domains (D1-D3) in the extracellular region. The present study aims to construct "off‑the‑shelf" CAR-NK cells targeting the membrane-proximal domain of CD5 derived from nanobody against T-cell malignancies.

Methods: Anti-CD5-D3 nanobody was screened by phage display technology, followed by constructing fourth-generation CAR plasmids ectopically producing IL-15 to generate CD5 CAR-NK cells derived from peripheral blood. And the second-generation CD5 CAR-T cells based on nanobody were generated, referred to as 5D.b CAR-T and 12 C.b CAR-T. Furthermore, CAR-NK cells without IL-15 (IL-15^△ CAR-NK) were generated to assess the impact on cytotoxicity of CAR-NK cells. Cytotoxic activity against CD5⁺ hematologic malignant cell lines and normal T cells was exerted in vitro and NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt mouse model transplanted with Jurkat-Luc cells was used to evaluate the antitumor efficacy of CD5 CAR-NK cells in vivo.

Results: Two nanobodies (5D and 12 C) competed for binding to the epitope of CD5-D3. 12 C CAR-NK cells were superior to 5D CAR-NK cells in antitumor potential and 12 C.b CAR-T cells exhibited superior cytotoxic activity than 5D CAR-T cells ex vivo. So, 12 C was regarded as the optimal nanobody. 12 C CAR-NK cells and IL-15^△ CAR-NK cells exhibited robust cytotoxicity against CD5⁺ malignant cell lines and controlled disease progression in xenograft mouse model. 12 C CAR-NK cells demonstrated greater antitumor activity compared to that of IL-15^△ CAR-NK cells in vitro and in vivo.

Conclusions: Taken together, the fourth-generation nanobody-derived anti-CD5 CAR-NK cells may be a promising therapeutic against T-cell malignancies.

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靶向 CD5 嵌合抗原受体工程化自然杀伤细胞，对抗 T 细胞恶性肿瘤。

背景：嵌合抗原受体工程T细胞（CAR-T）在B细胞恶性肿瘤中显示出良好的临床疗效，这种方法已被推广到T细胞恶性肿瘤中。然而，由于存在移植物抗宿主疾病的风险，在 CAR 疗法中使用异体 T 细胞是一项挑战。最近，自然杀伤（NK）细胞表现出了 "现成 "的可用性。基于纳米抗体的 CAR 结构因其治疗潜力而备受关注，这是因为纳米抗体具有体积小、稳定性最佳、亲和力强和生产可行性高等优点。CD5 是恶性 T 细胞常见的表面标志物，其细胞外区域有三个富含半胱氨酸的清道夫受体结构域（D1-D3）。本研究旨在构建 "现成的 "CAR-NK细胞，以纳米抗体衍生的CD5膜近端结构域为靶点，对抗T细胞恶性肿瘤：方法：通过噬菌体展示技术筛选出抗CD5-D3纳米抗体，然后构建异位产生IL-15的第四代CAR质粒，生成来自外周血的CD5 CAR-NK细胞。并生成了基于纳米抗体的第二代 CD5 CAR-T 细胞，分别称为 5D.b CAR-T 和 12 C.b CAR-T。此外，还生成了不含IL-15的CAR-NK细胞（IL-15△CAR-NK），以评估其对CAR-NK细胞细胞毒性的影响。在体外对CD5+血液恶性细胞系和正常T细胞发挥细胞毒活性，并用移植了Jurkat-Luc细胞的NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt小鼠模型评估CD5 CAR-NK细胞在体内的抗肿瘤功效：结果：两种纳米抗体（5D和12 C）竞争性地与CD5-D3的表位结合。12 C CAR-NK细胞的抗肿瘤潜力优于5D CAR-NK细胞，12 C.b CAR-T细胞在体内的细胞毒活性优于5D CAR-T细胞。因此，12 C 被认为是最佳的纳米抗体。12 C CAR-NK细胞和IL-15△CAR-NK细胞对CD5+恶性细胞株具有强大的细胞毒性，并能控制异种移植小鼠模型的疾病进展。与IL-15△CAR-NK细胞相比，12 C CAR-NK细胞在体外和体内表现出更强的抗肿瘤活性：综上所述，第四代纳米抗体衍生的抗 CD5 CAR-NK 细胞可能是一种治疗 T 细胞恶性肿瘤的有效方法。

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来源期刊

Experimental Hematology & Oncology Medicine-Oncology

CiteScore

12.60

自引率

7.30%

发文量

审稿时长

6 weeks

期刊介绍： Experimental Hematology & Oncology is an open access journal that encompasses all aspects of hematology and oncology with an emphasis on preclinical, basic, patient-oriented and translational research. The journal acts as an international platform for sharing laboratory findings in these areas and makes a deliberate effort to publish clinical trials with 'negative' results and basic science studies with provocative findings. Experimental Hematology & Oncology publishes original work, hypothesis, commentaries and timely reviews. With open access and rapid turnaround time from submission to publication, the journal strives to be a hub for disseminating new knowledge and discussing controversial topics for both basic scientists and busy clinicians in the closely related fields of hematology and oncology.