Targeting EphB4-ephrinB2 to decrease immunosuppression in HNSCC

靶向 EphB4-ephrinB2 减少 HNSCC 的免疫抑制

• 批准号: 10704596
• 负责人: SANA KARAM
• 金额: $ 32.47万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704596
• 关键词: Adhesions; Affect; Autoimmune; Beds; Biological Assay; Blood Vessels; Blood specimen; Cell Communication; Cell Proliferation; Cell Survival; Cell physiology; Cells; Clinical Trials; Coculture Techniques; Colorado; Cytometry; Cytoplasmic Tail; Cytoskeleton; Data; Disease; Embryonic Development; Endothelial Cells; Endothelium; Environment; EphB4 Receptor; Ephrin-B2; Flow Cytometry; Genetic; Genetically Engineered Mouse; Goals; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Human; Image; Immune; Immune Evasion; Immune Targeting; Immune checkpoint inhibitor; Immunocompetent; Immunofluorescence Immunologic; Immunofluorescence Microscopy; Immunologics; Immunosuppression; Immunotherapeutic agent; Implant; In Vitro; Infiltration; Intervention; L-Selectin; Ligands; Macrophage; Macrophage Activation; Malignant Neoplasms; Mediating; Mediator; Methods; Molecular; Molecular Target; Outcome; Participant; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Phosphorylation; Play; Population; Pre-Clinical Model; Process; Proto-Oncogene Proteins c-akt; Publishing; Radiation; Radiation therapy; Receptor Protein-Tyrosine Kinases; Regulatory T-Lymphocyte; Research; Resistance; Role; STAT3 gene; Sampling; Signal Transduction; Smoking History; Specimen; Stress Fibers; System; T cell response; Target Populations; Testing; Tissues; Tumor Promotion; Tumor-associated macrophages; Tumor-infiltrating immune cells; Vascular Endothelial Cell; angiogenesis; anti-tumor immune response; biomarker identification; cancer cell; cell motility; chemoradiation; cytokine; effector T cell; head and neck cancer patient; high risk; immunological status; immunoregulation; immunotherapy trials; in vivo; inhibitor; intravital microscopy; knock-down; migration; monocyte; neoplastic cell; novel; participant enrollment; pharmacologic; potential biomarker; pre-clinical; predictive marker; radiation response; radiological imaging; receptor; recruit; response; response biomarker; side effect; therapeutic target; therapy resistant; trafficking; transcriptome sequencing; treatment response; tumor; tumor growth; tumor microenvironment; tumor progression; tumor-immune system interactions

SUMMARY, Project 1 Treatment resistance remains a challenging problem for high-risk head and neck cancer (HNSCC) patients. Current trials with immune checkpoint inhibitors have yielded sub-optimal response rates. This could be due to TME factors that substantially modulate the immune landscape’s composition, phenotype, and function. Preferential recruitment of unique immunosuppressive cell subtypes to the tumor bed may play an important role in contributing to the promotion of tumor growth and progression. The tumor endothelium can act as a selective barrier that regulates entry, stability, and activation status of immune cells, but the mechanistic underpinnings remain poorly understood. The EphB4 receptor tyrosine kinase and its ligand EFNB2 define novel molecular targets. Although they have been studied in cell migration and angiogenesis in early embryonic development, little has been published on their role in modulating the cancer immune microenvironment. Our preliminary data show that HNSCCs are enriched in regulatory T-cells (Tregs) and tumor associated macrophages (TAMs), both of which contribute to resistance after RT. EFNB2 is upregulated on tumor vasculature after RT whereas EphB4 is expressed on Tregs and TAMs, and among all immune cells, Tregs and TAMs are most sensitive to either pharmacologic inhibition with TNYL-RAW or genetic knockdown of endothelial EFNB2 in the context of RT. We therefore hypothesize that RT upregulates EFNB2 on tumor vascular endothelial cells, which acts preferentially to recruit EphB4 expressing Tregs and TAMs. Blockade of EphB4-EFNB2 signaling at the tumor endothelial barrier will hinder Tregs’ and TAMs’ ability to infiltrate and promote cancer cell survival or suppress Teff function. In Aim1, we will analyze the mechanistic outcome of the interaction between immune cell EphB4 and endothelial EFNB2 on immune trans-endothelial trafficking, survival, polarization, and differentiation of immune cells as well as vascular normalization of endothelial cells in response to RT. We will use pharmacological inhibitors as well as GEMMs with EFNB2 deletion on endothelial cells or EphB4 deletion on Tregs or TAMs. Aim 2 will study the cellular and molecular mechanisms triggered by the interaction between endothelial EFNB2 and EphB4-expressing immune cells on immune cell processes in response to RT using endothelial-immune cells co-culture assays. Targeted inhibition of STAT3, AKT, and Erk pathway will be done with pharmacological inhibitors based on our preliminary data. Aim 3 will use tissue from a human clinical trial with sEphB4-HSA (an inhibitor currently in Phase I and II trials) before and after drug alone or drug and chemoradiation alone. This will serve to identify the molecular mediators that drive the anti-tumor immune response to this therapy. These studies will also collectively elucidate the molecular and cellular parameters of EphB4-EFNB2 inhibitors and will allow for the identification of potential predictive markers for this using EphB4- EFNB2 inhibitors.

项目1摘要 治疗耐药性仍然是高危头颈癌（HNSCC）患者面临的一个具有挑战性的问题。 目前使用免疫检查点抑制剂的试验产生了次优的应答率。这可能是由于 TME因子实质上调节免疫景观的组成、表型和功能。 独特的免疫抑制细胞亚型优先募集到肿瘤床可能发挥重要作用 有助于促进肿瘤生长和进展。肿瘤内皮细胞可以作为选择性的 调节免疫细胞进入、稳定和激活状态的屏障，但其机制基础 仍然知之甚少。EphB 4受体酪氨酸激酶及其配体EFNB 2定义新的分子 目标的虽然它们已经在早期胚胎发育中的细胞迁移和血管生成中进行了研究， 关于它们在调节癌症免疫微环境中的作用的报道很少。我们的初步数据 显示HNSCC富含调节性T细胞（TCFs）和肿瘤相关巨噬细胞（TAMs）， RT后，EFNB 2在肿瘤血管系统上上调，而EphB 4在肿瘤血管系统上上调。 在所有的免疫细胞中，T细胞和TAM对T细胞和TAM最敏感， 在RT的背景下，用TNYL-RAW或内皮EFNB 2的遗传敲低进行药理学抑制。 因此，我们假设RT上调肿瘤血管内皮细胞上的EFNB 2， 优先募集表达THB 4和TAM的EphB 4。EphB 4-EFNB 2信号传导在细胞中的阻断 肿瘤内皮屏障将阻碍TNM和TAM的浸润能力并促进癌细胞存活 或抑制Teff函数。在Aim 1中，我们将分析免疫系统之间相互作用的机制结果。 细胞EphB 4和内皮EFNB 2对免疫跨内皮运输、存活、极化和 免疫细胞的分化以及内皮细胞对RT的反应的血管正常化。 使用药理学抑制剂以及内皮细胞上EFNB 2缺失或EphB 4缺失的GEMM 在TMF或TAM上。目的2将研究细胞和分子机制触发的相互作用， 内皮EFNB 2和EphB 4表达免疫细胞对免疫细胞过程的影响 内皮-免疫细胞共培养测定。将进行STAT 3、AKT和Erk通路的靶向抑制 使用药物抑制剂来治疗。AIM 3将使用来自人体临床试验的组织 与sEphB 4-HSA（目前在I期和II期试验中的抑制剂）在单独药物或药物之前和之后， 单独放化疗这将有助于识别驱动抗肿瘤免疫的分子介质， 对这种疗法的反应。这些研究也将共同阐明的分子和细胞参数， EphB 4-EFNB 2抑制剂的作用，并将允许使用EphB 4-EFNB 2抑制剂鉴定潜在的预测标志物。 EFNB 2抑制剂。