Application of synthetic gene circuits to novel NK cell immunotherapy

我们建议使用合成生物学原理来开发癌症的下一代治疗方法，特别是癌症，并将重点放在卵巢癌(OC)上，以进行原理证明工作。包括CAR-T细胞和检查点抑制剂在内的几种不同形式的免疫疗法在治疗癌症方面取得了重大成功。然而，由于缺乏肿瘤抗原的可用性、T细胞耗竭或重大安全问题，如细胞因子风暴和自身免疫反应，这些方法对OC的治疗效果很少或根本没有。接入也受到成本和复杂性的限制。在这里，我们的目标是开发一个基于自然杀伤细胞(NK细胞)的通用癌症免疫治疗平台。这将基于一个通用的平台，以降低成本和增加可获得性，同时针对目前OC的糟糕结果。我们将增强和修改现有的基因电路模型，以通过NK细胞激活作为一种新的免疫疗法部署，它可以直接杀死肿瘤细胞。该电路将触发肿瘤细胞特异性分子的表达，这些分子可以增强肿瘤内内源性自然杀伤(NK)细胞的激活，或吸引更多NK细胞进入肿瘤。NK细胞在肿瘤早期清除中发挥关键作用，不需要预先存在的免疫或肿瘤抗原靶向。NK细胞治疗的临床潜力已经被证明(Shimasaki等人。《自然评论药物发现2020》)，没有毒性，也不需要后勤挑战和昂贵的自体生产，就像CAR-T细胞疗法的情况。我们将在Nissim等人2017年描述的基础上开发一种可适应的RNA介导的遗传逻辑电路。这种遗传电路的关键部件是一个布尔与门，它只有在两个输入信号同时存在的情况下才会产生输出信号。基因电路通过病毒载体转移到癌细胞中，在第一个要测试的病例中是卵巢癌细胞。只有一小部分卵巢癌细胞需要通过完整的回路进行转导，以产生输出，从而激活NK。这是该系统的一个主要USP。在临床上，只需要转导肿瘤中的一小部分癌细胞，就可以通过产生有效的输出分子来激活附近的NK细胞，从而产生显著的治疗效果。

We propose to use synthetic biology principles to develop a next-generation therapeutic approach for cancer, in particular carcinomas and focussing on Ovarian Cancer (OC) for our proof-of-principle work. Several different forms of immunotherapy including CAR-T cells and Checkpoint Inhibitors have had important success treating cancer. However, these approaches have had little or no therapeutic benefit for OC due to lack of tumour-antigen availability, T-cell exhaustion or major safety issues e.g. cytokine storms and autoimmune responses. Access is also limited by their cost and complexity. Here we aim to develop a universal platform for cancer immunotherapy based on Natural Killer (NK) cells. This will be based on a universal platform to lower cost and increase accessibility, whilst targeting the currently poor outcomes for OC.We will enhance and modify an existing gene circuit model to be deployed as a novel immunotherapy via NK cell activation, which can directly kill tumour cells. The circuit will trigger tumour cell-specific expression of molecules that can enhance the activation of endogenous Natural Killer (NK) cells within the tumour or attract more NK cells into the tumour. NK cells play a key role in early tumour clearance without requiring pre-existing immunity or tumour-antigen targeting. The clinical potential for NK cell therapy has already been demonstrated, (Shimasaki et al. Nature Reviews Drug Discovery 2020) without toxicity or need for logistically challenging and expensive autologous production, as in the case of CAR-T cell therapy.We will develop an adaptable RNA-mediated genetic logic circuit based on that previously described in Nissim et al., 2017. The key component of this genetic circuit is a Boolean AND gate that generates an output signal only if both input signals are present together. The genetic circuit is transferred by viral vectors into cancer cells, in the first case to be tested, ovarian cancer cells. Only a small proportion of the ovarian cancer cells need to be transduced with the complete circuit to lead to output and hence NK activation. This is a major USP of the system. In a clinical context, only a small proportion of the cancer cells within a tumour would need to be transduced to have a significant therapeutic effect by producing the potent output molecules to activate nearby NK cells.