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

项目摘要 我们将研究Tip 60的治疗靶向是否是一种组蛋白/蛋白乙酰转移酶， Foxp 3+调节性T细胞（Treg）的功能和存活可显著降低体内Treg功能， 保留常规的T细胞应答，导致在鼠模型中抑制肺肿瘤生长。我们将 还评估了Tip 60靶向对肿瘤内由常规药物诱导的DNA损伤反应的影响。 肺癌的治疗，包括放疗和顺铂治疗。我们开发了Tip 60 PROTAC （蛋白水解靶向嵌合）分子，其募集Cereblon E3连接酶以促进Tip 60 我们使用Cereblon敲入（CrbnI 391 V）免疫活性C57 BL/6小鼠， 筛选和肿瘤模型，以促进化合物的鉴定，用于随后的临床开发。 目的1 -Tip 60 i能否通过减少Foxp 3+来限制免疫活性宿主中的肿瘤生长 Treg抑制功能？我们已经开发了几种Tip 60 i，包括基于Crbn和VHL的PROTAC 化合物.我们将测试Tip 60 i PROTAC化合物是否可以调节Treg功能并控制肿瘤细胞的生长。 通过（1.1）优化Tip 60 i PROTAC化合物，并单独测试它们（1.2），或 在CrbnI 391 V小鼠中与（1.3）疫苗接种或（1.4）检查点抑制剂疗法联合。我们亦会评估 Tip 60 i对1.5）正常人Treg和/或Teff细胞，和1.6）人肺癌相关Treg的作用 细胞 目的2 -确定Tip 60 i是否破坏肿瘤细胞存活所必需的DDR作用。我们 分子研究将探索Tip 60依赖的机制，促进细胞对基因组的抗性， 不稳定性和正常的程序性死亡机制，这是癌症发生的基础， 化疗耐药性对肺癌细胞存活至关重要。我们将测试Tip 60 i化合物是否具有直接 通过破坏2.1）Tip 60依赖性p53活化和/或2.2）DNA修复机制来产生抗肿瘤作用。 我们的研究将促进恶性肿瘤患者免疫治疗新策略的发展， 鉴于新的Tip 60导向疗法具有双重作用机制，涉及靶向Foxp 3 + Treg， 细胞和内在肿瘤细胞对治疗的反应。所产生的数据可能会导致临床试验， 肺癌患者，并且还应该与其他个体相关，因为越来越多的 在许多其他类型的癌症的管理中，免疫疗法的公认潜力。

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.