Role of EphB4-ephrin-B2 interaction in regulating crosstalk between cancer cell and vascular tumor microenvironment in head and neck cancer

EphB4-ephrin-B2相互作用在头颈癌中调节癌细胞与血管肿瘤微环境之间的串扰中的作用

基本信息

批准号：
10565689
负责人：
SANA KARAM
金额：
$ 47.58万
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：
2019-03-01 至 2025-02-28
项目状态：
未结题

项目摘要

1 The outcomes for high-risk HPV-negative head and neck squamous cell carcinoma (HNSCC) remain poor 2 despite aggressive therapy. One possible explanation is the development of therapeutic resistance by aberrant 3 regulation of tumor angiogenesis. The receptor tyrosine kinase receptor, EphB4, and its membrane-bound 4 ligand, ephrinB2 (EFNB2), can both signal and have been shown to play pro-tumorigenic and pro-angiogenic 5 roles in numerous malignancies and in early embryonic development. Our data demonstrate strong correlation 6 between elevated gene expression of EphB4 and EFNB2 and survival outcomes in HPV negative HNSCC 7 patients. We show that EphB4 is predominantly expressed on cancer cells and EFNB2 is predominantly 8 present on endothelial cells (EC). Our data also show that targeting HNSCC tumors with a recombinant fusion 9 protein that blocks EphB4-EFNB2 interaction enhances cancer cell apoptosis, reduces 10 angiogenesis/lymphangiogenesis, and decreases tumor growth in patient-derived xenograft models. This, we 11 demonstrate, is associated with an increase in FasR, Bax, Bim, and caspase 3 cleavage apoptotic markers as 12 well as a decrease in VEGFR2/3, JAK2 and Stat3 signaling components. Based on these data, we hypothesize 13 that the interaction between EphB4 and EFNB2 at the cancer-EC junction simultaneously leads to activation of 14 survival and anti-apoptotic pathways for the cancer cell and angiogenic pathways for the EC. Therefore, we 15 propose, its inhibition will decrease tumor vascular/lymph angiogenesis and decreases cancer cell apoptosis. 16 This EphB4-EFNB2 interaction defines a novel cancer-EC “synapse,” where the focus is on the crosstalk and 17 not just on ECs, thus providing an opportunity for greater therapeutic efficacy than the currently available anti- 18 angiogenic therapies. In Aim 1, we will differentiate the compartmental effects of targeting the EphB4- 19 expressing cancer cell versus EFNB2-expressing ECs, by using an EFNB2 inducible knockout murine model 20 with selective deletion of EFNB2 in the adult ECs. Mouse HNSCC cell lines with shRNA and dominant negative 21 (DN) knockdown of EphB4 on cancer cells will be implanted and perfusion and immunofluorescence will be 22 used to assess effects on growth and angiogenesis/lymphangiogenesis. In Aims 2 and 3, in-depth interrogation 23 of signaling mechanisms underlying EphB4-EFNB2 interaction will be tested on in vivo tissue as well as 24 cancer-EC co-culture assays using shRNA and DN knockdowns for EphB4 and EFNB2. Targeted inhibition of 25 JAK2-STAT3-ERK and FasR, BAX, and PI3K will be done with genetic and pharmacologic inhibitors to assess 26 their role in vascular sprouting and cancer cell apoptosis. Proximity ligation assay will be done to evaluate 27 intracellular and intercellular interactions. Finally, tissue from the in vivo mouse model as well as from a clinical 28 trial incorporating an EphB4-EFNB2 inhibitor in HNSCC will be assessed using RNAseq, angiogenic, and 29 apoptotic arrays. Understanding the mechanistic underpinnings of interaction between the vasculature and 30 cancer cell will break down barriers that have stymied the angiogenesis field for decades.

1高风险HPV阴头和颈部鳞状细胞癌（HNSCC）的结果仍然很差 2目的地侵略性疗法。一种可能的解释是形成异常的理论抵抗力的发展 3调节肿瘤血管生成。受体酪氨酸激酶受体EPHB4及其膜结合 4配体Ephrinb2（EFNB2）都可以发出信号，并且已证明可以发挥促肿瘤和促血管生成作用在许多恶性肿瘤和早期胚胎发育中的5个角色。我们的数据表明相关性很强 6在HPV阴性HNSCC中，EPHB4和EFNB2的基因表达升高与生存结果之间 7例患者。我们表明，EPHB4主要在癌细胞上表达，EFNB2主要是 8存在于内皮细胞（EC）上。我们的数据还表明，针对重组融合的HNSCC肿瘤 9阻断EPHB4-EFNB2相互作用的蛋白质可增强癌细胞凋亡，减少 10血管生成/淋巴管生成，并降低患者衍生的纳学模型中的肿瘤生长。这个，我们 11证明，与FASR，BAX，BIM和caspase 3裂解凋亡标记的增加有关 12以及VEGFR2/3，JAK2和STAT3信号传导组件的减少。基于这些数据，我们假设 13癌症-EC连接处的EPHB4和EFNB2之间的相互作用只是导致激活 14癌细胞的生存和抗凋亡途径和EC的血管生成途径。因此，我们 15提案，其抑制作用将减少肿瘤血管/淋巴血管生成并减少癌细胞凋亡。 16这种EPHB4-EFNB2相互作用定义了一种新型的癌症-EC“突触”，其中重点是串扰和 17不仅在EC上，因此提供了比当前可用的反效率更高的治疗效率的机会 18种血管生成疗法。在AIM 1中，我们将区分靶向EPHB4-的隔室效应通过使用EFNB2诱导的敲除鼠模型，表达癌细胞与表达EFNB2的ECS 20在成人EC中有选择性删除EFNB2。小鼠HNSCC细胞系具有shRNA和显性阴性 EPHB4在癌细胞上的21（DN）将被植入，灌注和免疫荧光将是 22用于评估对生长和血管生成/淋巴管发生的影响。在目标2和3中，深入审讯 EPHB4-EFNB2相互作用的23个信号传导机制将在体内组织以及 24使用SHRNA和DN敲低的EPHB4和EFNB2的癌症-EC共培养分析。目标抑制 25 JAK2-STAT3-ERK和FASR，BAX和PI3K将使用遗传和药物抑制剂进行评估 26它们在血管发芽和癌细胞凋亡中的作用。接近连接测定将进行评估 27细胞内和细胞间相互作用。最后，来自体内小鼠模型以及临床的组织 28 HNSCC中的试验抑制剂将使用RNASEQ，血管生成和 29个凋亡阵列。了解脉管系统与 30癌细胞将打破数十年来阻碍血管生成场的障碍。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The interplay between cancer associated fibroblasts and immune cells in the context of radiation therapy.

DOI：
10.1002/mc.23205
发表时间：
2020-07
期刊：
Molecular carcinogenesis
影响因子：
4.6
作者：
Piper M;Mueller AC;Karam SD
通讯作者：
Karam SD

The role of concomitant chemoradiotherapy versus radiation alone in T1-3N0 HPV-positive and HPV-negative oropharyngeal squamous cell carcinoma.

DOI：
10.1016/j.oraloncology.2022.105907
发表时间：
2022-07
期刊：
ORAL ONCOLOGY
影响因子：
4.8
作者：
Shiao, Jay C.;Holt, Douglas;Ladbury, Colton;Gao, Dexiang;Jones, Bernard;Karam, Sana D.;Amini, Arya
通讯作者：
Amini, Arya

Tuberculosis-Cancer Parallels in Immune Response Regulation.