Harnessing the immunogenic potential of ferroptosis to improve immunotherapy in liver cancer

铁死亡是最近发现的一种程序性细胞死亡形式，在包括癌症在内的许多疾病中都是失调的，新的研究表明，铁死亡可能比其他类型的程序性细胞死亡更具免疫原性，这可能有助于癌症治疗。因此，我们的目标是研究铁下垂在肝癌中的免疫原性，这是迫切需要新的和更有效的治疗方法。原发性肝癌——主要是肝细胞癌（HCC）和肝内胆管癌（CCA）——是癌症相关死亡的第四大原因，通常发生在慢性炎症性肝病患者中。免疫疗法正在成为许多类型癌症的新治疗策略，但肝癌的反应率仍然很低。越来越清楚的是，免疫治疗的疗效高度依赖于炎症性肿瘤表型与肿瘤反应性T细胞的高浸润。因此，能够将未发炎的肿瘤微环境转化为发炎程度更高的肿瘤微环境的治疗方法有很大的潜力来提高免疫治疗的疗效。先前的研究表明，诱导免疫原性细胞死亡，其特征是释放可以激活免疫系统的介质，可以诱导强烈的肿瘤特异性免疫反应。铁下垂是一种由于铁积累和氧化应激而发生的程序性细胞死亡，越来越多的证据表明铁下垂可能具有免疫原性潜力，可以作为改善免疫治疗的目标。虽然参与铁下垂调节的分子途径和行为者已经被彻底研究，但免疫细胞在铁下垂背景/环境中的作用以及铁下垂如何影响免疫系统仍然知之甚少。本研究旨在描述铁下垂对免疫系统的影响，特别是在抗癌免疫治疗的背景下。我们认为，肝肿瘤细胞的铁下垂可以诱导肿瘤特异性免疫反应，并且这种免疫系统的激活将与肝癌免疫治疗协同作用并增加（目前不足的）疗效。我们将使用体外培养系统和肝癌小鼠模型来表征铁下垂引起免疫反应的机制。铁下垂引起的免疫变化将采用创新的多重技术（光谱流式细胞术，成像细胞术）进行分析。基于这些结果，我们还将把铁下垂诱导与其他免疫疗法（如检查点阻断）结合起来。最后，我们将使用肝癌患者的肝脏和血清样本验证临床前结果，以便将结果转化为临床试验。我们的研究结果将确定如何将铁下垂纳入针对HCC和CCA的治疗策略。

Ferroptosis is a recently identified form of programmed cell death that is dysregulated in many diseases including cancer, and emerging studies indicate that ferroptosis might be more immunogenic than other types of programmed cell death, which could prove beneficial in cancer treatment. Therefore, we aim to study the immunogenicity of ferroptosis in the context of liver cancer, where new and more efficient therapies are urgently needed. Primary liver cancer - mainly hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (CCA) - is the fourth leading cause of cancer-related death and typically occurs in patients with chronic inflammatory liver diseases. Immunotherapies are emerging as new treatment strategies for many types of cancer, but the response rates in liver cancer are still very low. It has become increasingly clear that efficacy of immunotherapy highly depends on an inflamed tumor phenotype with high infiltration of tumor-reactive T cells. Therefore, therapies that can convert an uninflamed tumor microenvironment to a more inflamed environment have a strong potential to boost efficacy of immunotherapy. Prior work suggests that induction of immunogenic cell death, which is characterized by the release of mediators that can activate the immune system, can induce strong tumor-specific immune responses. Ferroptosis is a type of programmed cell death that occurs as a result of iron accumulation and oxidative stress and there is increasing evidence that ferroptosis might have immunogenic potential and could be targeted to improve immunotherapy. While the molecular pathways and actors involved in the regulation of ferroptosis have been thoroughly investigated, the role of immune cells in a ferroptotic context/environment and how ferroptosis can influence the immune system remains poorly understood. This proposal aims to characterize the influence of ferroptosis on the immune system, especially in the context of anticancer immunotherapy. We propose that ferroptosis of hepatic tumor cells can induce a tumor-specific immune response and that this activation of the immune system will synergize with and increase the (currently insufficient) efficacy of immunotherapy in liver cancer. We will use in vitro culture systems and mouse models of liver cancer to characterize the mechanisms by which ferroptosis elicits an immune response. The immunological changes induced by ferroptosis will be analyzed using innovative multiplex technologies (spectral flow cytometry, imaging mass cytometry). Based on the results, we will also combine ferroptosis induction with other immunotherapies, such as checkpoint blockade. Finally, we will validate the pre-clinical results using human liver and serum samples from patients with liver cancer to facilitate the translation of the results to clinical trials. The results of our studies will establish how ferroptosis may be incorporated into therapeutic strategies against HCC and CCA.