Tackling Treg mediated resistance to radiation and anti-PDL1 in HNSCCs

解决 HNSCC 中 Treg 介导的放射抗性和抗 PDL1

基本信息

批准号：
10590767
负责人：
SANA KARAM
金额：
$ 43.91万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10590767
关键词：
Adjuvant Radiotherapy Affect Animal Model Back Biological Assay Blood Blood specimen CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene CXC chemokine receptor 3 CXCL9 gene CXCR3 gene Cell secretion Cells Chemotaxis Clinical Trials Design Coculture Techniques Cytometry Data FOXP3 gene Feeds Flow Cytometry Genetic Engineering Genetic Transcription Genetically Engineered Mouse Goals Growth Growth Factor Head and Neck Cancer Head and Neck Squamous Cell Carcinoma Human Immune Immune Evasion Immune Targeting Immune checkpoint inhibitor Immunofluorescence Immunologic Immunotherapy In Vitro Infiltration Interferon Type II Investigational Therapies Knockout Mice Knowledge Laboratories LoxP-flanked allele Mediating Modeling Molecular Mus Neoadjuvant Therapy PD-L1 blockade Pathway interactions Patients Phase Phase I Clinical Trials Phenotype Phosphorylation Plasma Proliferating Radiation Radiation therapy Regulatory T-Lymphocyte Relapse Research Research Proposals Resistance Resistance development STAT1 gene STAT3 gene Signal Transduction Signaling Molecule T-Cell Activation T-Cell Depletion TCR Activation Testing Tissue Sample Tissues Up-Regulation anti-CTLA4 anti-PD-L1 anti-PD-L1 therapy anti-tumor immune response cancer cell carcinogenesis checkpoint therapy chemokine clinical trial analysis cofactor cytokine effective therapy effector T cell first-in-human genome sequencing high risk immune checkpoint blockade improved inhibitor inhibitor therapy knock-down migration mouse model negative affect novel therapeutic intervention pharmacologic phase I trial polarized cell potential biomarker programmed cell death ligand 1 recruit response synergism targeted treatment therapy development therapy resistant transcriptome sequencing tumor tumor eradication tumor microenvironment whole genome

项目摘要

Abstract Current treatment of head and neck squamous cell carcinoma (HNSCC) with immune checkpoint inhibitors and radiotherapy (RT) are initially effective, but relapse is common. The goal of our research is to understand mechanisms of response and resistance and to use this knowledge to improve therapy. We show that during the response phase to treatment, RT increases STAT1 phosphorylation, CXCL9/10 secretion, and PDL1 expression on cancer cells, and increases CD8 and CD4 infiltration and activation. These effects, however, dissipate in the resistance phase, and a significant increase in regulatory T cells (Tregs), STAT3, and TGF1 are observed. Treg depletion and anti-STAT3 targeting restores response to RT and anti-PDL1, resulting in suppression of plasma levels of TGF1, activation of T effector cells (Teff, Cd44+, IFN-G+ CD4 and CD8 T cells) and complete tumor eradication in an orthotopic model of HNSCC. This effect is unachievable with the addition of anti-CTLA4, anti-Tim3, or with hypofractionation of RT. Our in vitro preliminary data show that RT enhances STAT3 phosphorylation on CD4 T cells, leading to increased Treg conversion. This is reversed with STAT3 inhibitors. CD4 T cell conversion toward a Treg phenotype represents an important mechanism of immune evasion. TGF1 has been established as a necessary growth factor for conversion of naïve CD4 T cells to Tregs through induction of FoxP3 expression. In addition, STAT3 is a necessary transcription co-factor for FoxP3 expression. Our central hypothesis is that during the response phase, RT induces a distinct cancer cell STAT1-mediated CXCL9/10 secretion which recruits and activates Teff resulting in enhanced synergy with anti-PDL1. RT, however, also promotes CD4 conversion to Treg cells by phosphorylation of STAT3, which leads to the development of treatment resistance by inhibiting Teff function. Using knockdown constructs of STAT1 and CXCL9/10 on cancer cells and a CXCR3-/- mouse model, we will determine the effect of RT-induced chemokines in Teff chemotaxis, proliferation, and TCR activation (Aim1). To determine how RT affects CD4 T cell conversion and how that feeds back to negatively affect Teff function, we will use the CD4-Cre/ Stat3flox/flox mice with knockdown of STAT3 on CD4 T cells as well as STAT3 inhibitors (Aim2). Finally, we will use tissue and blood samples from a Phase I clinical trial aimed at using neoadjuvant combination RT and anti-PDL1 and will examine whether that changes the immune landscape to increase Teff/Treg ratio and enhance Teff activation profile (Aim3). We expect these studies to elucidate molecular and cellular parameters of RT plus immune checkpoint blockade and to help develop more effective therapy for HNSCC.

抽象的目前使用免疫检查点抑制剂和免疫检查点抑制剂治疗头颈鳞状细胞癌（HNSCC）放射治疗 (RT) 最初是有效的，但复发很常见，我们研究的目的是了解这一点。我们证明了反应和抵抗机制，并利用这些知识来改进治疗。在治疗反应阶段，RT 会增加 STAT1 磷酸化、CXCL9/10 分泌和 PDL1 癌细胞上的表达，并增加 CD8 和 CD4 的浸润和激活。抵抗期消散，调节性 T 细胞 (Treg)、STAT3 和 TGF1 显着增加观察到 Treg 耗竭和抗 STAT3 靶向可恢复对 RT 和抗 PDL1 的反应，从而导致抑制血浆 TGF1 水平，激活 T 效应细胞（Teff、Cd44+、IFN-G+ CD4 和 CD8 T）细胞）并在 HNSCC 原位模型中完全根除肿瘤，这种效果是 HNSCC 无法实现的。添加抗 CTLA4、抗 Tim3 或联合大分割 RT 我们的体外初步数据表明 RT。增强 CD4 T 细胞上的 STAT3 磷酸化，导致 Treg 转化增加。 STAT3 抑制剂是 CD4 T 细胞向 Treg 表型转化的重要机制。 TGF1 已被确定为幼稚 CD4 T 转化的必要生长因子。细胞通过诱导 FoxP3 表达形成 Tregs 此外，STAT3 是必要的转录辅助因子。我们的中心假设是，在反应阶段，RT 会诱导明显的变化。癌细胞 STAT1 介导的 CXCL9/10 分泌招募并激活 Teff，从而导致然而，与抗 PDL1 RT 的增强协同作用也可促进 CD4 转化为 Treg 细胞。 STAT3 磷酸化，通过抑制 Teff 导致治疗耐药性的产生在癌细胞和 CXCR3-/- 小鼠上使用 STAT1 和 CXCL9/10 的敲低构建体。模型中，我们将确定 RT 诱导的趋化因子对 Teff 趋化性、增殖和 TCR 的影响确定 RT 如何影响 CD4 T 细胞转化以及它如何产生负面影响。影响 Teff 功能，我们将使用 CD4-Cre/ Stat3flox/flox 小鼠，同时敲低 CD4 T 细胞上的 STAT3 最后，我们将使用来自 I 期临床试验的组织和血液样本。使用新辅助组合 RT 和抗 PDL1，并将检查这是否会改变免疫景观以增加 Teff/Treg 比率并增强 Teff 激活曲线（目标 3）。阐明放疗加免疫检查点阻断的分子和细胞参数，并帮助开发更多 HNSCC 的有效治疗方法。