Improvement of NK cell-infusion therapy for metastatic breast cancer by targeting tumour-associated macrophages

通过靶向肿瘤相关巨噬细胞改善转移性乳腺癌的 NK 细胞输注疗法

• 批准号: MR/S006982/1
• 负责人: Takanori Kitamura
• 金额: $ 140.61万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FS006982%2F1
• 关键词: Improvement; NK; cell; infusion; therapy

Breast cancer is one of the leading causes of cancer death in women. Although most cancer in the breast (primary tumours) can be resected surgically, it often spreads to the different part of the body such as the bone and lung (metastatic tumours) and this is responsible for the high mortality of the disease. Since existing therapies are not sufficient to prolong survival of patients with metastatic breast cancer, novel approaches are required to improve outcome. An emerging therapeutic strategy is the infusion of immune cells called natural killer (NK) cell that can directly kill certain types of cancer cells. Clinical and pre-clinical studies have shown that infusion of NK cells exerts modest therapeutic effects in breast cancer patients without adverse side effects. However, several reports show that NK cells cannot exhibit full anti-tumour ability in the environment of the tumour, suggesting that elimination of such a suppressive environment is needed to use the NK cells as a therapeutic tool. Recent studies suggest that other types of immune cells called tumour-associated macrophages (TAMs) can suppress NK cell functions in the primary tumours. However, their roles in the metastatic tumours are largely unknown. Using animal models of metastatic breast cancer, I have reported that TAMs are one of the most abundant cell types in the environment of metastatic tumours, and their accumulation helps cancer cells to establish the lethal metastatic tumours. Furthermore, my recent studies suggest that the TAMs in the metastatic tumours can suppress NK cell-induced tumour cell death in vitro. We have also found that macrophages acquire NK cell suppressive functions when they are stimulated with conditioned media that are harvested from cultured cancer cells and thus include several factors secreted by cancer cells. I thus predict that the TAMs in the metastatic tumor environment are educated by cancer cells to prevent therapeutic effects of the infused NK cells on the metastatic breast cancer. The overarching ambition of my project is to improve efficacy of the NK cell-based therapy against metastatic breast cancer by blocking functions of TAMs. To achieve this ultimate goal, this proposal has three specific aims. The first aim is to define how TAMs suppress NK cell functions. We will isolate TAMs from murine metastatic tumors, and investigate their effects on NK cell movement, activation, killing factor expression, and tumor killing activity using in vitro assays established by us. The second aim is to identify therapeutic targets that can potentiate human NK cell efficacy. We will identify candidate factors that are highly expressed in human macrophages educated by cancer cells, and assess whether ablation of these factors prevent the macrophage mediated suppression of the NK cell functions. We will also define the cancer cell-derived factors that educate human macrophages to be NK suppressive cells. The third aim is to develop clinically relevant models for validation of the therapeutic targets and further drug discovery. We have established a protocol to differentiate human induced pluripotent stem cell (iPSC) into mature macrophages and NK cells. Using these cells, we will develop our in vitro assay into a more clinically relevant model. We will also establish a mouse model by which we can assess therapeutic effects of human NK cell infusion combined with blockade of identified therapeutic targets on the metastasis formation of human breast cancer cells. I believe that this research will show us the essential factors for TAMs to suppress NK cell functions, which will lead us to develop novel approaches to improve efficacy of NK cell-based therapy for metastatic diseases. The funding from this award would allow me to expand my research group, enhance my independence, develop further collaborations, and thereby identify new strategies that will potentially prolong survival of metastatic breast cancer patients.

乳腺癌是女性癌症死亡的主要原因之一。虽然大多数乳腺癌（原发性肿瘤）可以通过手术切除，但它通常会扩散到身体的不同部位，如骨和肺（转移性肿瘤），这是导致该疾病高死亡率的原因。由于现有的治疗方法不足以延长转移性乳腺癌患者的生存期，因此需要新的方法来改善结果。一种新兴的治疗策略是输注称为自然杀伤（NK）细胞的免疫细胞，可以直接杀死某些类型的癌细胞。临床和临床前研究表明，输注NK细胞在乳腺癌患者中发挥适度的治疗作用，没有不良副作用。然而，一些报告显示NK细胞在肿瘤环境中不能表现出完全的抗肿瘤能力，这表明需要消除这种抑制性环境以使用NK细胞作为治疗工具。最近的研究表明，称为肿瘤相关巨噬细胞（TAM）的其他类型的免疫细胞可以抑制原发性肿瘤中的NK细胞功能。然而，它们在转移性肿瘤中的作用在很大程度上是未知的。使用转移性乳腺癌的动物模型，我已经报道了TAM是转移性肿瘤环境中最丰富的细胞类型之一，它们的积累有助于癌细胞建立致命的转移性肿瘤。此外，我最近的研究表明，转移性肿瘤中的TAM可以抑制体外NK细胞诱导的肿瘤细胞死亡。我们还发现，当巨噬细胞用条件培养基刺激时，它们获得NK细胞抑制功能，所述条件培养基从培养的癌细胞收获，因此包括癌细胞分泌的几种因子。因此，我预测转移性肿瘤环境中的TAM被癌细胞训练以阻止输注的NK细胞对转移性乳腺癌的治疗作用。我的项目的总体目标是通过阻断TAM的功能来提高基于NK细胞的治疗对转移性乳腺癌的疗效。 为了实现这一最终目标，本建议有三个具体目标。第一个目标是确定TAM如何抑制NK细胞功能。我们将从小鼠转移性肿瘤中分离TAM，并使用我们建立的体外试验研究它们对NK细胞运动、活化、杀伤因子表达和肿瘤杀伤活性的影响。第二个目的是确定可以增强人NK细胞功效的治疗靶点。我们将鉴定在受癌细胞训练的人巨噬细胞中高度表达的候选因子，并评估这些因子的消融是否阻止巨噬细胞介导的NK细胞功能抑制。我们还将定义癌细胞衍生的因子，这些因子使人巨噬细胞成为NK抑制细胞。第三个目标是开发临床相关模型，用于验证治疗靶点和进一步的药物发现。我们已经建立了一种将人诱导多能干细胞（iPSC）分化为成熟巨噬细胞和NK细胞的方案。使用这些细胞，我们将把我们的体外试验发展成更临床相关的模型。我们还将建立一种小鼠模型，通过该模型我们可以评估人NK细胞输注联合阻断已鉴定的治疗靶点对人乳腺癌细胞转移形成的治疗效果。我相信这项研究将向我们展示TAM抑制NK细胞功能的基本因素，这将引导我们开发新的方法来提高基于NK细胞的治疗转移性疾病的疗效。该奖项的资助将使我能够扩大我的研究小组，增强我的独立性，发展进一步的合作，从而确定可能延长转移性乳腺癌患者生存期的新策略。

项目成果

Tumour-associated macrophages as a potential target to improve natural killer cell-based immunotherapies.

• DOI: 10.1042/ebc20230002
• 发表时间: 2023-09-28
• 期刊: Essays in biochemistry
• 影响因子: 6.4

Enzyme-Activatable Chemokine Conjugates for In Vivo Targeting of Tumor-Associated Macrophages

用于体内靶向肿瘤相关巨噬细胞的酶激活趋化因子缀合物

• DOI: 10.1002/ange.202207508
• 发表时间: 2022
• 期刊: Angewandte Chemie
• 作者: Barth N

Enzyme-Activatable Chemokine Conjugates for In Vivo Targeting of Tumor-Associated Macrophages.

• DOI: 10.1002/anie.202207508
• 发表时间: 2022-10-10
• 期刊: ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
• 影响因子: 16.6
• 作者: Barth, Nicole D.;Van Dalen, Floris J.;Karmakar, Utsa;Bertolini, Marco;Mendive-Tapia, Lorena;Kitamura, Takanori;Verdoes, Martijn;Vendrell, Marc
• 通讯作者: Vendrell, Marc

Fluorogenic Granzyme A Substrates Enable Real-Time Imaging of Adaptive Immune Cell Activity

荧光颗粒酶 A 底物可实现适应性免疫细胞活性的实时成像

• DOI: 10.1002/ange.202216142
• 发表时间: 2023
• 期刊: Angewandte Chemie
• 作者: Cheng Z

Fluorogenic Granzyme A Substrates Enable Real-Time Imaging of Adaptive Immune Cell Activity.

• DOI: 10.1002/anie.202216142
• 发表时间: 2023-02-13
• 期刊: Angewandte Chemie (International ed. in English)