A Novel ULK1 Pathway as a New Therapeutic Target in Melanoma

项目摘要/摘要 黑色素瘤是一种高度致命的恶性肿瘤，免疫治疗方法已被证明对其有效。 虽然免疫检查点阻断(ICB)改变了晚期黑色素瘤的治疗，但大多数 患者最初对治疗有抵抗力，或有反应，但后来复发。因此，努力找出 对检查点抑制剂的耐药性以及克服原发和获得性耐药性的方法是必要的。 令人惊讶的是，II型干扰素信号通路和干扰素刺激基因(ISGs)的表达 黑色素瘤患者已被证明与ICB治疗的反应或抵抗有关，在一项 依赖于上下文的方式。而肿瘤反应性T细胞的效应功能需要干扰素诱导的基因 在黑色素瘤中，干扰素也可诱导免疫抑制基因的表达 慢性干扰素信号增强获得性耐药。干扰素信号的这种双重作用支持 需要确定有选择地调节其作用以促进细胞毒性T淋巴细胞(CTL)活性的方法 而不会促进免疫抑制。我们最近发现ULK1(UNC-51-like kinase1)控制 特异性干扰素信号事件的激活和特异性干扰素诱导基因的转录参与控制 免疫反应。重要的是，我们的初步研究表明，ULK1的基因或药物靶向 在黑色素瘤细胞中抑制干扰素诱导的免疫抑制基因的表达而不影响表达 免疫刺激因子的表达和ULK1的高水平表达与黑色素瘤的低存活率相关 病人。该项目的目标是确定靶向ULK1是否能阻断干扰素依赖的免疫抑制。 在促进干扰素介导的细胞毒性T细胞杀伤黑色素瘤活性的同时，促进其效应和相关的生物学反应。目标1 将明确ULK1在干扰素诱导的免疫抑制基因表达和细胞毒作用中的特定作用 T淋巴细胞抗黑色素瘤活性。它包括产生特定的敲除黑色素瘤细胞，以及 ULK体外抑制T细胞介导的肿瘤杀伤作用的评价目标2将定义ULK1在ICB中的作用- 黑色素瘤中介导的免疫反应。基因突变或基因突变对黑色素瘤生长的影响 ULK1与免疫检查点抑制剂的联合药理抑制作用将在体内进行检验。 此外，ULK1的表达和激活将与黑色素瘤患者的临床反应相关 接受ICB治疗。这个项目的结果将促进我们对特定的干扰素信号如何 这些事件影响针对黑色素瘤的免疫反应的成功，并将为新的靶向提供基础 干扰素信号转导的特异性调节剂与免疫检查点抑制剂联合治疗的方法 黑色素瘤的治疗。

Project Summary/Abstract Melanoma is a highly fatal malignancy for which immunotherapy approaches have been shown to be effective. While immune checkpoint blockade (ICB) has transformed the treatment of advanced melanoma, the majority of patients are resistant to therapy initially or respond but then relapse. Thus, efforts to identify mechanisms of resistance to checkpoint inhibitors and approaches to overcome primary and acquired resistance are warranted. Surprisingly, type II interferon (IFN) signaling pathways and expression of IFN-stimulated genes (ISGs) in melanoma patients have been shown to correlate with either response or resistance to ICB treatment, in a context-dependent manner. While IFN-inducible genes are required for effector function of tumor-reactive T cells and response to ICB in melanoma, IFN is also known to induce expression of immunosuppressive genes and chronic IFN signaling promotes acquired resistance to ICB. This dual role of IFN signaling supports the need to identify means to selectively regulate its effects in order to promote cytotoxic T lymphocyte (CTL) activity without promoting immunosuppression. We have recently discovered that ULK1 (Unc-51-like kinase 1) controls activation of unique IFN signaling events and transcription of specific IFN-induced genes involved in the control of immune responses. Importantly, our preliminary studies show that genetic or pharmacologic targeting of ULK1 in melanoma cells represses IFN-induced expression of immunosuppressive genes without affecting expression of immunostimulatory ones, and high levels of ULK1 expression correlate with poor survival in melanoma patients. The goal of this project is to determine if targeting ULK1 blocks IFN-dependent immunosuppressive effects and related biological responses, while promoting IFN-mediated CTL activity against melanoma. Aim 1 will define the specific role of ULK1 on IFN-induced expression of immunosuppressive genes and on cytotoxic T lymphocyte activity against melanoma. It includes generation of specific knockout melanoma cells, and evaluation of ULK inhibition on T-cell mediated tumor killing in vitro. Aim 2 will define the role of ULK1 in ICB- mediated immune responses in melanoma. The effects on melanoma tumor growth of genetic or pharmacological inhibition of ULK1 in combination with immune checkpoint inhibitors will be examined in vivo. Additionally, the expression and activation of ULK1 will be correlated to clinical responses in melanoma patients receiving ICB therapy. The results of this project will advance our understanding on how specific IFN signaling events affect the success of immune responses against melanoma and will provide the basis for novel targeted approaches involving combination of specific regulators of IFN signaling with immune checkpoint inhibitors for the treatment of melanoma.