蛋白酶体抑制增强嵌合抗原受体(CAR)表达的 NK 细胞对急性髓性白血病的抗白血病疗效

IF 29.5 1区 医学 Q1 HEMATOLOGY
David Sedloev, Qian Chen, Julia M. Unglaub, Nicola Schanda, Yao Hao, Eleni Besiridou, Brigitte Neuber, Anita Schmitt, Simon Raffel, Yi Liu, Maike Janssen, Carsten Müller-Tidow, Michael Schmitt, Tim Sauer
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引用次数: 0

摘要

复发性和难治性急性髓性白血病(AML)的预后很差。CAR T细胞对急性髓性白血病的疗效有限,部分原因是自体T细胞功能失调,以及患者特异性CAR T细胞的生成时间延长。异体 NK 细胞疗法是一种很有前景的替代疗法,但可能需要采取一些策略来提高疗效和持久性。蛋白酶体抑制剂(PI)会诱导表面蛋白质组发生变化,这可能会使恶性细胞更容易受到 NK 介导的细胞毒性的影响。在此,我们研究了将蛋白酶抑制剂与 CAR 表达的异体 NK 细胞结合治疗急性髓细胞性白血病的潜在益处。我们确定了硼替佐米和卡非佐米对几种急性髓细胞白血病细胞系的 IC50 浓度。多参数流式细胞术测定了蛋白酶体抑制剂处理后I类HLA分子和应激相关蛋白的表面表达。利用功能性体外试验,我们探索了PIs预处理与表达或不表达AML特异性CAR构建体的NK细胞对AML细胞系和原发性患者样本的抗白血病疗效之间的协同作用。此外,我们还研究了在两种不同的急性髓细胞白血病小鼠异种移植模型中应用单一 PI 策略,然后输注(CAR-)NK 细胞的耐受性和疗效。急性髓细胞白血病细胞系和原发性急性髓细胞白血病患者样本对硼替佐米和卡非佐米介导的细胞毒性易感。对阿扎胞苷/Venetoclax的条件耐药性不会产生对PIs的原发性耐药性。用PIs治疗急性髓细胞性白血病细胞可减少急性髓细胞性白血病细胞表面I类HLA分子的表达,这种表达具有时间和剂量依赖性。应激相关蛋白在转录水平和细胞表面上调。NK 细胞介导的对 AML 细胞的杀伤以协同方式得到增强。PI预处理增加了效应细胞-靶细胞共轭物的形成和干扰素-γ的分泌,从而增强了NK细胞在体外对AML细胞系和原代样本的活性。CD33和CD70特异性CAR的表达进一步提高了抗白血病的疗效。在体内,硼替佐米(Bortezomib)预处理后输注 CAR-NK 细胞可减少急性髓细胞白血病的生长,从而延长总生存期。PIs增强了CAR表达的异体NK细胞在体外和体内对AML的抗白血病疗效,值得在早期临床试验中进一步探索这种组合疗法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Proteasome inhibition enhances the anti-leukemic efficacy of chimeric antigen receptor (CAR) expressing NK cells against acute myeloid leukemia
Relapsed and refractory acute myeloid leukemia (AML) carries a dismal prognosis. CAR T cells have shown limited efficacy in AML, partially due to dysfunctional autologous T cells and the extended time for generation of patient specific CAR T cells. Allogeneic NK cell therapy is a promising alternative, but strategies to enhance efficacy and persistence may be necessary. Proteasome inhibitors (PI) induce changes in the surface proteome which may render malignant cells more vulnerable to NK mediated cytotoxicity. Here, we investigated the potential benefit of combining PIs with CAR-expressing allogeneic NK cells against AML. We established the IC50 concentrations for Bortezomib and Carfilzomib against several AML cell lines. Surface expression of class-I HLA molecules and stress-associated proteins upon treatment with proteasome inhibitors was determined by multiparameter flow cytometry. Using functional in vitro assays, we explored the therapeutic synergy between pre-treatment with PIs and the anti-leukemic efficacy of NK cells with or without expression of AML-specific CAR constructs against AML cell lines and primary patient samples. Also, we investigated the tolerability and efficacy of a single PI application strategy followed by (CAR-) NK cell infusion in two different murine xenograft models of AML. AML cell lines and primary AML patient samples were susceptible to Bortezomib and Carfilzomib mediated cytotoxicity. Conditioned resistance to Azacitidine/Venetoclax did not confer primary resistance to PIs. Treating AML cells with PIs reduced the surface expression of class-I HLA molecules on AML cells in a time-and-dose dependent manner. Stress-associated proteins were upregulated on the transcriptional level and on the cell surface. NK cell mediated killing of AML cells was enhanced in a synergistic manner. PI pre-treatment increased effector-target cell conjugate formation and Interferon-γ secretion, resulting in enhanced NK cell activity against AML cell lines and primary samples in vitro. Expression of CD33- and CD70-specific CARs further improved the antileukemic efficacy. In vivo, Bortezomib pre-treatment followed by CAR-NK cell infusion reduced AML growth, leading to prolonged overall survival. PIs enhance the anti-leukemic efficacy of CAR-expressing allogeneic NK cells against AML in vitro and in vivo, warranting further exploration of this combinatorial treatment within early phase clinical trials.
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来源期刊
CiteScore
48.10
自引率
2.10%
发文量
169
审稿时长
6-12 weeks
期刊介绍: The Journal of Hematology & Oncology, an open-access journal, publishes high-quality research covering all aspects of hematology and oncology, including reviews and research highlights on "hot topics" by leading experts. Given the close relationship and rapid evolution of hematology and oncology, the journal aims to meet the demand for a dedicated platform for publishing discoveries from both fields. It serves as an international platform for sharing laboratory and clinical findings among laboratory scientists, physician scientists, hematologists, and oncologists in an open-access format. With a rapid turnaround time from submission to publication, the journal facilitates real-time sharing of knowledge and new successes.
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