PROTAC Targeting of Tip60 in Syngeneic Cancer Models Using Cereblon Knock-in Mice

项目摘要 我们将研究Tip60的治疗靶向，Tip60是一种组蛋白/蛋白乙酰转移酶，对 Foxp3+T细胞调节性T细胞(Treg)的功能和存活在体内可显著降低Treg功能 保存传统的T细胞反应，导致抑制小鼠肺癌模型的生长。我们会 也评估了Tip60靶向对常规药物诱导的肿瘤内DNA损伤反应的影响 肺癌的治疗，包括放射治疗和顺铂治疗。我们已经开发了Tip60 PROTAC (蛋白水解靶向嵌合体)招募Cereblon E3连接酶以促进Tip60的分子 降解，并采用Cereblon基因敲除(CrbnI391V)免疫活性C57BL/6小鼠在化合物 筛选和肿瘤模型，以促进化合物的鉴定，以便随后的临床开发。 目的1-Tip60i能通过降低Foxp3+抑制免疫活性宿主肿瘤的生长 特雷格抑制功能？我们已经开发了几个Tip60i，包括基于CRBN和VHL的PROTAC 化合物。我们将测试Tip60i PROTAC化合物是否可以调节Treg功能并控制肿瘤的生长 通过(1.1)优化Tip60i PROTAC化合物，并单独(1.2)测试它们，或在 联合(1.3)疫苗接种或(1.4)检查点抑制剂治疗CrbnI391V小鼠。我们还将评估 Tip60i对正常人Treg细胞和/或Tef细胞以及人肺癌相关Treg细胞的影响 细胞。 目的2-确定Tip60i是否干扰对肿瘤细胞生存至关重要的DDR动作。我们的 分子研究将探索Tip60依赖的促进细胞对基因组耐药的机制 不稳定和正常的程序性死亡机制，是癌症发生和发展的基础 化疗耐药性对肺癌细胞的生存至关重要。我们将测试Tip60i化合物是否有直接 通过干扰依赖于Tip60的P53激活和/或DNA修复机制发挥抗肿瘤作用。 我们的研究将促进恶性肿瘤患者免疫治疗新策略的开发， 鉴于新的Tip60导向疗法具有双重作用机制，涉及靶向Foxp3+Treg 细胞和固有的肿瘤细胞对治疗的反应。将产生的数据可能会导致在 肺癌患者，也应该与其他人相关，因为越来越多的人 认识到免疫疗法在许多其他类型的癌症治疗中的潜力。

Project Summary We will investigate whether therapeutic targeting of Tip60, a histone/protein acetyltransferase essential for the functions and survival of Foxp3+ T-regulatory (Treg) can significantly decrease Treg functions in vivo while preserving conventional T cell responses, leading to inhibition of lung tumor growth in murine models. We will also assess the effects of Tip60 targeting on the DNA damage response within tumors induced by conventional therapies for lung cancer, including irradiation and cisplatin therapy. We have developed Tip60 PROTAC (proteolysis targeting chimeric) molecules that recruit the Cereblon E3 ligase so as to promote Tip60 degradation, and we employ Cereblon knock-in (CrbnI391V) immunocompetent C57BL/6 mice in compound screening and tumor models to facilitate identification of compounds for subsequent clinical development. Aim 1 - Can Tip60i limit growth of tumors in immunocompetent hosts by decreasing Foxp3+ Treg suppressive function? We have developed several Tip60i, including Crbn- and VHL-based PROTAC compounds. We will test if Tip60i PROTAC compounds can modulate Treg function and control growth of experimental lung cancers by (1.1) optimizing Tip60i PROTAC compounds, and testing them (1.2) alone, or in conjunction with (1.3) vaccination or (1.4) checkpoint inhibitor therapy in CrbnI391V mice. We will also assess effects of Tip60i on 1.5) normal human Treg and/or Teff cells, and 1.6) human lung cancer associated Treg cells. Aim 2 – Determine if Tip60i disrupts DDR actions essential for tumor cell survival. Our molecular investigations will explore Tip60-dependent mechanisms that promote cellular resistance to genome instability and normal programmed mechanisms of death, and which underpin cancer initiation and chemotherapeutic resistance crucial for lung cancer cell survival. We will test if Tip60i compounds have direct anti-tumor effects by disrupting 2.1) Tip60-dependent p53 activation and/or 2.2) DNA repair mechanisms. Our studies will facilitate the development of new strategies for immunotherapy in patients with malignancies, given that the new Tip60-directed therapies have dual mechanisms of action, involving targeting Foxp3+ Treg cells and intrinsic tumor cell responses to therapy. The data to be generated will likely lead to clinical trials in patients with lung cancers, and should also have relevance to additional individuals, given the increasingly recognized potential for immunotherapy in the management of many other types of cancers.