A novel murine model of squamous lung cancer

鳞肺癌（SQC）是一种非小细胞肺癌（NSCLC）。目前还没有特定的治疗药物被许可用于治疗SQC，晚期患者的预后非常差。对于另一种常见类型的非小细胞肺癌——肺腺癌（ADC），有一些靶向治疗方法，而且对其驱动的分子事件有更多的了解。同样，也有肺腺癌的小鼠模型，这对理解其生物学非常有帮助，并且可以与临床疾病直接相关。与人类类器官方法相比，小鼠模型的一个主要优势是，它们可以用来理解肿瘤发展和进展及其微环境之间的动态和复杂关系。临床迫切需要改进我们的SQC方法，这可以通过一个合理的小鼠模型来极大地帮助，该模型可以复制肿瘤发展的遗传和组织微环境背景。迄今为止的努力已经取得了一些成功，但不幸的是，它们在转化研究或测试潜在治疗组合方面的效用有限。为了解决这一未满足的需求，我们开发了一种制作SQC小鼠模型的新方法。我们使用来自动物肺的小鼠细胞，并在组织培养中扩增它们，然后使用实验室技术对它们进行操作，以重现在人类疾病中看到的遗传异常。然后，我们将这些细胞注射回基因相同（近亲繁殖）的小鼠的肺部，这些细胞导致SQC肿瘤。该议定书对3r的任务有重大影响。一个原因是，对于这种疾病的传统遗传模型来说，需要许多育种才能获得正确的基因组合来发展癌症类型。在这种情况下，基因是在细胞中而不是在动物中进行操作的，因此该过程使用的老鼠明显更少，技术上也更容易。第二个原因是，我们将细胞移植到肺的一部分，这样动物就不会出现明显的不适，而只会长出局部肿瘤。这将使我们能够完善协议，从而大大减少对动物福利的总体影响。第三，许多现有的SQC模型使用毒素产生潜在复杂且严重副作用的SQC。在这个建议中，我们不寻求在任何阶段对任何小鼠施用毒素。最后，我们的目标是改造这些细胞，这样我们就可以在单个动物身上监测疾病和治疗的影响。这些措施的进一步综合影响是在SQC上做小鼠实验的成本显著降低。我们已经在试点数据中证明，我们的方法在3周内产生早期小鼠SQC病变，在野生型免疫能力动物中大约4个月产生大的侵袭性肿瘤。我们的目标是验证和复制我们的试点数据，并证明其与人类疾病的直接相关性。此外，我们将证明该模型可以有效地用于解决关键的基础和转化问题：癌细胞保持存活的特定驱动癌基因的必要性；使用药物预防鳞状肺癌发展的潜力；这个模型将用于研究肿瘤如何与其微环境相互作用。我们对这个项目的可能性感到兴奋，并热衷于与该方法在学术界和制药行业的潜在最终用户进行沟通，以确保该模型得到广泛应用。

Squamous lung cancer (SQC) is a type of non-small cell lung cancer (NSCLC). There are currently no specific therapeutic agents licensed to treat SQC and outcomes for patients with advanced disease are very poor. There are some targeted therapeutics available for the other common type of NSCLC - lung adenocarcinoma (ADC) - and more is understood about the molecular events that drive it. Likewise, there are mouse models of lung adenocarcinoma available that have been very helpful in understanding its biology and that can have direct relevance to the clinical disease. A major advantage that mouse models have over human organoid approaches is that they can be used to understand the dynamic and complex relationship between the developing and progressing tumour and its microenvironment. There is an urgent clinical need to improve our approach to SQC and this could be helped enormously by a rational mouse model that can replicate the genetic and tissue microenvironment context in which the tumour develops. Efforts to date have had some successes but, unfortunately, they are of limited utility in terms of translational studies or testing potential therapeutic combinations.In order to address this unmet need we have developed a new way of making a mouse model of SQC. We use mouse cells from an animal's lung and expand them in tissue culture before manipulating them using laboratory techniques to recreate the genetic abnormalities seen in the human disease. We then inject these cells back into the lung of a genetically identical (inbred) mouse and these cells lead to SQC tumours.This protocol has a major impact on the mission of the 3Rs. One reason is that for the traditional genetic models of this disease many breedings are required to get the right combination of genes to develop the type of cancer. In this case the genes are manipulated in cells rather than in an animal, so the process uses markedly fewer mice and is technically facile. A second reason is that we deliver the cells to one part of the lung, so that the animals develop a local tumour without becoming overtly unwell. This should allow us to refine the protocol so that the overall impact on animal welfare is significantly lessened. Third, many of the existing models of SQC use toxins to generate SQC with potentially complex and serious side effects. In this proposal we do not seek to administer toxins to any mouse at any stage. Finally we aim to engineer these cells so that we can monitor disease and the impact of a treatment in a single animal. A further combined impact of these measures is a marked reduction in the cost of doing mouse experiments on SQC.We have demonstrated in pilot data that our approach generates early murine SQC lesions within 3 weeks and large invasive tumours at around 4 months in wild type immunocompetent animals. We aim to validate and reproduce our pilot data and demonstrate its direct relevance to the human disease. Further we will show that the model can be used efficiently to address key basic and translational issues: the necessity of a specific driving oncogene for the cancer cells to stay alive; the potential to use drugs to prevent squamous lung cancer development; and a demonstration that this model will have utility for studying how the tumour interacts with its microenvironment.We are excited at the possibilties of this project and are enthusiastic about communicating with the potential end-users of this approach in academic community and pharma to ensure that the model is widely used.