Natural killer cells: of-the-shelf cytotherapy for cancer immunosurveillance

Natural killer (NK) cells are advantaged innate cytotoxic lymphocytes with characteristics of tumor immunosurveillance and microorganism elimination. Distinguish from the adaptive T and B lymphocytes, the autologous or allogeneic NK cells efficaciously fulfil the function of combating transformed hematological malignancies and metastatic solid tumors via the proverbial mechanisms including direct cytolytic effect and antibody-dependent cell-mediated cytotoxicity (ADCC) as well as paracrine effects dispense with antigen presentation. Herein, we review the candidate sources (e.g., peripheral blood, umbilical cord blood, placental blood, cell lines and stem cells) for large-scale and clinical-grade NK cell manufacturing, ex vivo cultivation (feeder-, cytokine cocktail- or physicochemical irritation-dependent strategies) for NK cell persistence and activation. Furthermore, we also figure out the promising prospects as well as the accompanied challenges of NK cell- or chimeric antigen receptor-transduced NK (CAR-NK) cell-based adoptive immunotherapy in standardizations for industrialized preparation and clinical practices.     

超声实时引导下对晚期直肠癌患者进行NKG2D-CAR基因修饰的NK细胞瘤内局部注射:一例病例报道

嵌合抗原受体NK细胞免疫治疗（Chimeric Antigen Receptor NK-Cell Immunotherapy，CAR-NK）是目前最具潜力的抗肿瘤治疗方式之一，而改善CAR-NK细胞的回输方式是提高CAR-NK细胞抗肿瘤作用的重要途径，现报道一例实时超声引导下的瘤内注射在对晚期直肠癌患者进行NKG2D-CAR基因修饰的NK细胞回输中的应用。     

Chimeric Antigen Receptor-Natural Killer Cells: A New Breakthrough in the Treatment of Solid Tumours

Natural killer (NK) cells can quickly and directly eradicate tumour cells without recognising tumour-specific antigens. NK cells also participate in immune surveillance, which arouses great interest in the development of novel cancer therapies. The chimeric antigen receptor (CAR) family is composed of receptor proteins that give immune cells extra capabilities to target specific antigen proteins or enhance their killing effects. CAR-T cell therapy has achieved initial success in haematological tumours, but is prone to adverse reactions, especially with cytokine release syndrome in clinical applications. Currently, CAR-NK cell therapy has been shown to successfully kill haematological tumour cells with allogeneic NK cells in clinical trials without adverse reactions, proving its potential to become an off-the-shelf product with broad clinical application prospects. Meanwhile, clinical trials of CAR-NK cells for solid tumours are currently underway. Here we will focus on the latest advances in CAR-NK cells, including preclinical and clinical trials in solid tumours, the advantages and challenges of CAR-NK cell therapy and new strategies to improve the safety and efficacy of CAR-NK cell therapy.     

CAR-NK抗肿瘤研究的现状与发展趋势

作为固有免疫中重要的效应细胞,NK细胞具有强大的抗肿瘤功能,在肿瘤免疫治疗方面具有良好的应用前景.随着嵌合抗原受体(chimeric antigen receptor,CAR)修饰的T细胞(CAR-T)技术的兴起,NK细胞的CAR修饰潜力也受到了关注.CAR-NK的研发既传承了经典CAR-T研发的思路,也实现了诸多基于NK细胞生物特点的创新和发展.外周血NK细胞、NK细胞系和干细胞来源的NK细胞各具生物学特色,在CAR-NK研发中被广泛使用.已有靶向多种血液肿瘤和实体瘤抗原的CAR-NK在体外实验和动物模型中取得显著效果.虽然在临床暂未有数据支持CAR-NK的疗效且仍存在一些瓶颈问题需要攻克,但是CAR-NK有望在治疗实体瘤、简化免疫治疗等方面实现新的突破.     

龙生蛭胶囊联合阿加曲班治疗急性脑梗死的临床研究

目的探讨龙生蛭胶囊联合阿加曲班治疗急性脑梗死的临床疗效。方法选取2019年1月—2020年4月天津市中医药大学附属南开中医院收治的急性脑梗死患者146例,随机分为对照组和治疗组,每组各73例。对照组静脉滴注阿加曲班注射液,60 mg加入生理盐水500 mL,持续24 h,3~7 d后调整为20 mg加入生理盐水100 mL,持续2~3 h,2次/d。治疗组患者在对照组的基础上,口服龙生蛭胶囊治疗,5粒/次,疗程为2周。观察两组患者临床疗效,比较治疗前后两组患者生活质量、神经功能缺损评分(NIHSS)、Barthel指数(BI)评分、颈动脉斑块,及钙调素(CAM)、肿瘤坏死因子-α(TNF-α)、正五聚体蛋白-3(PTX-3)、生长分化因子-15(GDF-15)和白细胞介素-6(1L-6)水平。结果治疗后,治疗组的总有效率为93.15%,明显高于对照组的79.45%(P<0.05)。治疗后,两组生理功能、情感功能和社会功能评分较治疗前的明显升高(P<0.05),治疗组升高更明显(P<0.05)。两组患者在治疗的第7天、第14天,NIHSS评分与BI评分较治疗前明显改善(P<0.05),治疗组在治疗的第14天,NIHSS评分与BI评分明显好于对照组(P<0.05);与对照组相比,治疗组IMT厚度和斑块面积降低更为显著(P<0.05)。治疗后,两组CAM、TNF-α、PTX-3、GDF-15和1L-6水平较治疗前显著降低(P<0.05),而且与对照组相比,治疗组下降更为明显(P<0.05)。结论龙生蛭胶囊联合阿加曲班治疗急性脑梗死效果显著,能够有效的提高患者的生活质量,降低神经功能缺损的严重程度和斑块面积,有效的改善临床指标。     

CAR-NK cell in cancer immunotherapy; A promising frontier

Chimeric antigen receptors (CARs) have a unique facet of synthetic biology and offer a paradigm shift in personalized medicine as they can use and redirect the patient's immune cells to attack cancer cells. CAR-natural killer (NK) cells combine the targeted specificity of antigens with the subsequent intracellular signaling ability of the receptors to increase their anti-cancer functions. Importantly, CAR-NK cells can be utilized as universal cell-based therapy without requiring human leukocyte antigen (HLA) matching or earlier contact with tumor-associated antigens (TAAs). Indeed, CAR-NK cells can be adapted to recognize various antigens, hold higher proliferation capacity, and in vivo persistence, show improved infiltration into the tumors, and the ability to overcome the resistant tumor microenvironment leading to sustained cytotoxicity against tumors. Accumulating evidence from recent in vivo studies rendering CAR-NK cell anti-cancer competencies renewed the attention in the context of cancer immunotherapy, as these redirected effector cells can be used in the development of the “off-the-shelf” anti-cancer immunotherapeutic products. In the current review, we focus on the therapeutic efficacy of CAR-NK cell therapies for treating various human malignancies, including hematological malignancies and solid tumors, and will discuss the recent findings in this regard, with a special focus on animal studies.     

基因修饰体细胞国内外临床研究和相关产业现状及未来发展趋势

基因编辑技术可针对性增强体细胞的靶向性,一定程度上解决了传统过继性免疫疗法在肿瘤等多种疾病治疗中显露出的靶向性差等问题。最具代表性的是嵌合抗原受体T细胞(CAR-T)免疫疗法,其在血液系统肿瘤治疗中已取得良好疗效,此外,T细胞受体基因工程化的T细胞(TCR-T)、基因修饰的树突状细胞以及基因修饰的干细胞在多种疾病治疗的临床研究中也显现了良好的安全性和有效性。目前国内外多家研究机构和生物公司已在基因修饰体细胞行业加速布局,相关产业即将进入快速发展期,基因修饰体细胞疗法的临床转化应用具有广阔前景,对目前已经开展疾病临床研究的几种基因修饰免疫细胞疗法进行总结。     

人源IL-15转基因NCG小鼠在CAR-NK细胞肿瘤治疗临床前评价中的应用

目的：探讨人源IL-15转基因NCG小鼠（NCG-hIL-15鼠）在CAR-NK细胞肿瘤治疗临床前评价中的作用。方法：通过qPCR结合WB法检测NCG-hIL-15鼠骨髓及主要器官（脾、肝、肺、肾和胰）中的表达丰度。将人 PBMC 来源的NK（PB-NK）细胞回输到NCG-hIL-15鼠及对照NCG鼠,监测NK细胞的体内扩增与受体鼠的体质量、生存期变化,流式细胞术检测体内扩增NK细胞的活化受体与抑制受体表达差异 ,WB 法检测穿孔素与颗粒酶 B 的表达。MIAPaca-2 细胞原位荷瘤NCG-hIL-15鼠或NCG鼠用anti-MUC1-CAR-NK细胞回输治疗,流式细胞术检测CAR-NK细胞在不同组鼠体内的存活情况,小动物活体成像术检测肿瘤生长并记录荷瘤鼠生存期。结果：NCG-hIL-15鼠中PB-NK细胞可以在10周内稳定增殖,且观察期内未发生显著的移植物抗宿主反应。NCG-hIL-15鼠中扩增NK细胞的KAR与KIR丰度与原始细胞相比差异较小;与NCG鼠体内增殖的NK细胞相比,NCG-hIL-15鼠体内扩增的NK细胞有更高水平的穿孔素与颗粒酶B表达（均P<0.05）。CAR-NK细胞在NCG-hIL-15鼠体内存活比显著增加、抑瘤效果增强,与对照组相比NCG-hIL-15鼠生存期显著延长（均P<0.01）。结论：NCG-hIL-15鼠模型可用于基于NK细胞免疫疗法的临床前实验与生物学评价。     

Current State of the Art of Allogeneic CAR Approaches – Pile ‘Em High and Sell ’Em Cheap

The advent and rapid propagation of Chimeric Antigen Receptor (CAR)-based therapeutics in recent years has taken the oncology field by storm and delivered considerable benefit to cancer patients, many of whom previously had no other treatment options available to them. CAR-based therapies are now a bona fide therapeutic modality in the fight against cancer, along with more “traditional” treatments, such as small molecule and antibody drugs. For the technology to take the next step and reach much larger numbers of patients in need, it will be necessary for those treatments to become “off-the-shelf” offering patients a standardised, consistent, and cost-effective product. This article offers an overview of the evolution and development options for off-the-shelf CAR-based treatments, the advantages and disadvantages of the various approaches, along with key optimisation parameters that must be considered.     

CAR-NK的构建策略及在血液系统肿瘤中的研究进展北大核心

自然杀伤(natural killer cells,NK)细胞是先天免疫系统不可或缺的一部分,为抵御异常细胞攻击人体提供了第一道屏障。嵌合抗原受体T细胞(CAR-T)工程是近年来癌症免疫治疗领域的研讨热点,它改变了血液肿瘤的传统治疗方法,这种新颖的治疗方式已经在血液肿瘤中显示出了令人振奋的结果。然而,CAR-T的临床应用受到了高昂的制造成本和各种副作用的限制。NK细胞由于具有更好的安全性、“现成”生产的可行性和强大的细胞毒性激发了人们开发用于癌症治疗的CAR工程NK细胞(CAR-NK)的兴趣,CAR-NK有望取代CAR-T成为更有前景的抗癌方法。本文中我们描述了当前CAR-NK的构建策略和在血液系统肿瘤中的研究进展。