Determining the role of the SDF-1/CXCR4 pathway and its intersection with chronic stress to establish novel precision approaches to head and neck cancer management

项目摘要/摘要 大量临床前证据表明，在头部动物模型中，应激诱导肿瘤进展 和宫颈癌(HNC)。临床上，局部晚期人乳头瘤病毒(HPV)阴性的患者 HNC暴露在广泛的慢性应激源中。癌症进展与慢性应激之间的联系 可能是HPV阴性的HNC的肿瘤学结果持续较差的原因之一 人口。为了评估慢性应激的机制和发展治疗方法，需要建立慢性应激的动物模型。 然而，现有的模型要么使用免疫缺陷小鼠，要么使用同基因小鼠肿瘤，这两种模型都不是 概述人类肿瘤微环境(TME)。为了克服这些限制，我们开发了一部小说 慢性应激小鼠模型设计成具有人类免疫系统并携带人HNC异种移植。 在使用该模型的初步研究中，慢性应激导致肿瘤生长和相关转移的增加。 循环基质衍生因子-1(SDF-1)水平升高和TME的免疫改变。 综上所述，我们假设慢性应激导致SDF-1/CXCR4途径上调，从而诱导 免疫耐受的TME和增加HNC的肿瘤生长和转移。为了检验这一假说， 确定临床相关性，我们将研究应激诱导的肿瘤生长的机制。 细胞来源的异种移植(CDX)和患者来源的异种移植(PDX)人源化的HNC模型。我们将进一步 通过对接受HNC手术的患者进行前瞻性观察研究，评估临床相关性。 在目标1中，人源化的CDX小鼠将被随机分为慢性社会隔离应激或对照条件。三 将设计的ARM包括(1)用CXCR4拮抗剂治疗的动物，(2)用CXCR4治疗的HNC异种移植 缺失，以及(3)带有野生型HNC异种移植的对照臂。肿瘤的生长和肺转移将 量过了。将进行肿瘤内免疫表型和RNA测序。在目标2中，我们将招募 局部晚期HPV阴性HNC患者计划接受手术切除，患有 预后不佳，长期应激源的基线暴露。患者将接受血浆细胞因子 评估和肿瘤切除标本将接受免疫表型和RNA测序。在子集中 在这些患者中，将产生自体人源化患者衍生异种移植(PDX)模型，在该模型中 人类的免疫系统和肿瘤来自同一个患者。这对于HNC手术来说是唯一可行的 随着腓骨重建，作为一段腓骨被常规丢弃，其中的造血干细胞 可以收获用于自体人性化。这些自体PDX模型将被随机分配到慢性 应激和控制条件，并用CXCR4拮抗剂处理与对照注射相比较。 这项研究将是第一次在SDF的背景下研究应激诱导的HNC生长的机制。 1/CXCR4轴和人TME。来自人性化临床前模型和平行前瞻性的结果 临床分析将为精确疗法的发展和合理的临床试验设计奠定基础。

PROJECT ABSTRACT/SUMMARY Substantial pre-clinical evidence has demonstrated stress-induced tumor progression in animal models of head and neck cancer (HNC). Clinically, patients with locoregionally advanced Human Papillomavirus (HPV)-negative HNC are exposed to a broad range of chronic stressors. The link between cancer progression and chronic stress may be one contributing explanation for the persistently poor oncologic outcomes of the HPV-negative HNC population. To evaluate mechanism and develop therapeutics, animal models of chronic stress are needed. However, existing models use either immunodeficient mice or syngeneic murine tumors, neither of which recapitulate the human tumor microenvironment (TME). To overcome these limitations, we developed a novel mouse model of chronic stress engineered to have a human immune system and carry human HNC xenografts. In preliminary studies using this model, chronic stress led to increased tumor growth and metastases associated with higher levels of circulating stromal-derived factor 1 (SDF-1) and immune alterations in the TME. Taken together, we hypothesize that chronic stress leads to SDF-1/CXCR4 pathway upregulation which induces an immunotolerant TME and increased tumor growth and metastasis in HNC. To test this hypothesis and determine clinical relevance, we will investigate the mechanisms underlying stress-induced tumor growth in both cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) humanized models of HNC. We will further evaluate clinical relevance through a prospective observational study in patients undergoing HNC surgery. In Aim 1, humanized CDX mice will be randomized to chronic social isolation stress or control conditions. Three arms will be designed including (1) animals treated with a CXCR4 antagonist, (2) HNC xenografts with a CXCR4 deletion, and (3) a control arm with wild-type HNC xenografts. Tumor growth and lung metastases will be measured. Intra-tumoral immunophenotyping and RNA sequencing will be performed. In Aim 2, we will recruit patients with locoregionally advanced HPV-negative HNC planned for surgical resection, a population with a poor prognosis and significant baseline exposure to chronic stressors. Patients will undergo plasma cytokine assessment and tumor resection specimens will undergo immunophenotyping and RNA sequencing. In a subset of these patients, autologous humanized patient-derived xenograft (PDX) models will be generated, in which the human immune system and tumor are derived from the same patient. This is uniquely feasible for HNC surgery with fibula reconstruction, as a segment of fibula bone is routinely discarded from which hematopoietic stem cells can be harvested for autologous humanization. These autologous PDX models will be randomized to chronic stress and control conditions and treated with a CXCR4 antagonist as compared with control injection. This study will be the first to investigate the mechanism of stress-induced HNC growth in the context of the SDF- 1/CXCR4 axis and the human TME. Results from the humanized pre-clinical models and parallel prospective clinical analyses will lay the foundation for development of precision therapeutics and rational clinical trial design.