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Regulation of Zbtb44-Eomes complex in CD8+T cells and anti-tumor immunity

CD8 T 细胞中 Zbtb44-Eomes 复合物的调节和抗肿瘤免疫

基本信息

批准号：
10574602
负责人：
Venuprasad K Poojary
金额：
$ 41.01万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-24 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10574602
关键词：
Binding CAR T cell therapy CD8-Positive T-Lymphocytes CD8B1 gene CRISPR/Cas technology Cancer Model Carcinoembryonic Antigen Cell Therapy Cell physiology Cells Clinical Trials Colon Carcinoma Complex Dissection Equilibrium Event Family Functional disorder Gene Expression Profile Immune Immune checkpoint inhibitor Interferons MC38 Mediating Molecular Mus Pathway interactions Patients Play Post-Translational Protein Processing Regulation Role Solid Neoplasm Sumoylation Pathway T cell response T cell therapy Testing Tumor Immunity Tumor-Infiltrating Lymphocytes Ubiquitination anti-CTLA4 antibodies anti-PD-1 cancer therapy chimeric antigen receptor chimeric antigen receptor T cells exhaust exhaustion improved outcome member novel patient derived xenograft model prevent programmed cell death protein 1 promoter protein complex protein degradation therapeutic evaluation therapeutic target transcription factor tumor tumor growth tumor microenvironment ubiquitin ligase

项目摘要

ABSTRACT CAR-T cell therapy is an emerging option for cancer treatment, but its efficacy is limited, especially in solid tumors because the effector CD8+T cells become dysfunctional and exhausted in the tumor microenvironment (TME). However, the key pathways that define the delicate balance between the effector vs exhausted state of CD8+T cells remain unclear. Our preliminary studies demonstrate that sumoylation of the T-box transcription factor, Eomesodermin (Eomes), facilitates its association with Zbtb44, a member of the ThPOK family of transcription factors. The Zbtb44-Eomes complex promotes the effector function and anti-tumor activity of CD8+ tumor infiltrated lymphocytes (TILs). In exhausted CD8+ TILs, the ubiquitin ligase Trim47 targets Zbtb44 for degradation and disrupts the Zbtb44-Eomes complex. Furthermore, CRISPR-Cas9-mediated inhibition of Trim47 rescues exhausted CD8+ TILs and restores their effector function. These preliminary findings led us to hypothesize that ubiquitination and sumoylation of the Zbtb44/Eomes complex are critical molecular events that dictate the effector vs exhaustion of CD8+ TILs which can be therapeutically targeted. In Aim1, we will determine the mechanism by which the Zbtb44-Eomes complex promotes effector CD8+T cell function and anti-tumor immunity. We will use newly generated Zbtb44-/- mice to investigate how sumoylation of Eomes at Lys(K)-446 facilitates the formation of the Zbtb44-Eomes complex via the SUMO interacting motif (SIM) within Zbtb44. Further, we will delineate the mechanism by which the Zbtb44-Eomes complex cooperatively binds to and transactivates the IFN- promoter. In Aim 2, we will determine the mechanism by which Trim47-mediated ubiquitination of Zbtb44 leads to dysfunction of CD8+T cells. We will investigate how Trim47, which is upregulated in exhausted (PD1+Tim3+) CD8+ TILs, targets Zbtb44 for ubiquitination at K139 and promotes its degradation. Using newly generated Trim47-/- mice, we will determine how disruption of the Zbtb44- Eomes complex leads to the inhibitory transcriptional profile of exhausted CD8+T cells. In Aim 3, we will target the Zbtb44-Trim47 pathway to promote anti-tumor immunity. We will test the therapeutic potential of blocking Zbtb44 ubiquitination in CAR-T cells against carcinoembryonic antigen (CEA) in the MC38 and in a patient- derived xenograft (PDX) colon cancer model. Completion of these studies will lead to: 1) dissection of the novel Zbtb44-Eomes complex that is critical for effector CD8+ T cell function, 2) determination of how Trim47-mediated ubiquitination disrupts this complex leading to alternate transcription profile in exhausted CD8+ TILs, and 3) evaluate the means to target the Zbtb44- Trim47 pathway to overcome the current limitations of CAR-T cell therapy for solid tumors.

摘要 CAR-T细胞疗法是癌症治疗的新兴选择，但其疗效有限，尤其是在实体瘤中因为效应CD 8 +T细胞在肿瘤微环境（TME）中变得功能失调并耗尽。然而，定义效应细胞与CD 8 +T细胞耗竭状态之间微妙平衡的关键途径，细胞仍然不清楚。我们的初步研究表明，T-box转录因子Eomesodermin（Eomes）的sumoylation，促进其与Zbtb 44（转录因子的ThPOK家族的成员）的缔合。Zbtb44-Eomes 复合物促进CD 8+肿瘤浸润淋巴细胞（TIL）的效应子功能和抗肿瘤活性。在当CD 8 + TIL耗尽时，泛素连接酶Trim 47靶向Zbtb 44降解并破坏Zbtb 44-Eomes 复杂.此外，CRISPR-Cas9介导的Trim 47的抑制拯救了耗尽的CD 8 + TIL并恢复了CD 8 + TIL。其效应器功能。这些初步的发现使我们假设，泛素化和类小泛素化的蛋白质， Zbtb 44/Eomes复合物是决定CD 8 + TIL的效应子与耗竭的关键分子事件其可以是治疗靶向的。在Aim 1中，我们将确定Zbtb 44-Eomes复合物促进效应CD 8 +T细胞增殖的机制。功能和抗肿瘤免疫。我们将使用新产生的Zbtb 44-/-小鼠来研究类小泛素化是如何影响小鼠的免疫应答的。赖氨酸（K）-446位的Eomes通过SUMO相互作用基序促进Zbtb 44-Eomes复合物的形成 (SIM)在ZBTB 44中。此外，我们将描述Zbtb 44-Eomes复合物协同结合并反式激活IFN-γ启动子。在目标2中，我们将通过以下方式确定机制： Trim 47介导的Zbtb 44泛素化导致CD 8 +T细胞功能障碍。我们将调查如何 Trim 47在耗尽的（PD 1 + Tim 3+）CD 8 + TIL中上调，靶向Zbtb 44在K139处的泛素化，促进其退化。使用新产生的Trim 47-/-小鼠，我们将确定如何破坏Zbtb 44-/-小鼠。 Eomes复合物导致耗尽的CD 8 +T细胞的抑制性转录谱。在目标3中，我们将针对 Zbtb 44-Trim 47通路促进抗肿瘤免疫。我们将测试阻断在MC 38和患者中CAR-T细胞中针对癌胚抗原（CEA）的Zbtb 44泛素化衍生的异种移植物（PDX）结肠癌模型。这些研究的完成将导致：1）对新的Zbtb 44-Eomes复合物的解剖，该复合物对于效应CD 8 + T细胞功能，2）确定Trim 47介导的泛素化如何破坏该复合物导致耗尽的CD 8 + TIL中的替代转录谱，和3）评估靶向Zbtb 44-TIL的方法， Trim 47通路克服目前CAR-T细胞治疗实体瘤的局限性。