Sexual dimorphism and the immuno-modulatory role of estrogen signaling in HNSCC

HNSCC 中雌激素信号的性别二态性和免疫调节作用

• 批准号: 10751465
• 负责人: SANA KARAM
• 金额: $ 64.63万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-03 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751465
• 关键词: Adhesions; Affect; Agonist; Antigen Presentation; Antigen Presentation Pathway; Antigens; Autoimmunity; Automobile Driving; Biological; Biological Assay; Biological Markers; Blood; Bone Marrow; Castration; Cell Cycle Kinetics; Cell Differentiation process; Cell Line; Cell Survival; Cell physiology; Cells; Cellular biology; Chimera organism; Circulation; Clinical Trials; Coculture Techniques; Data; Dendritic Cells; Dependence; Disease; Dose; Endosomes; Estradiol; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogen Receptors; Estrogens; FLT3 ligand; Female; Flow Cytometry; Fluorescent Antibody Technique; Genetic; Genetically Engineered Mouse; Goals; Gonadal Steroid Hormones; Head and Neck Cancer; High Endothelial Venule; Homing; Hormones; Human; Immune; Immune response; Immunosuppression; Immunotherapeutic agent; Immunotherapy; In Vitro; Incidence; Inflammation; Knockout Mice; Label; Lymphocyte; Macrophage; Male Castration; Mediating; Mediator; Menopausal Status; Migration Assay; Modeling; Molecular; Mus; Nodal; Outcome; Ovalbumin; Ovariectomy; Patients; Phenotype; Play; Population; Postmenopause; Premenopause; Process; Proteomics; Receptor Signaling; Regulatory T-Lymphocyte; Reporting; Research; Risk; Role; Selection for Treatments; Sex Preselection; Signal Transduction; Site; System; T cell differentiation; T-Cell Activation; T-Lymphocyte; Target Populations; Testing; Testosterone; Therapeutic; Time; Toll-like receptors; Tomatoes; Tumor Antigens; Up-Regulation; Woman; antagonist; antigen-specific T cells; cancer cell; cancer therapy; cellular targeting; density; draining lymph node; effector T cell; experimental study; head and neck cancer patient; immunoregulation; improved; in vitro Assay; in vivo; lymphatic vessel; male; men; menopausal hormone therapy; mouse model; negative affect; novel marker; novel therapeutics; pharmacologic; pre-clinical; programmed cell death protein 1; receptor; response; restraint; sex; sexual dimorphism; therapy resistant; trafficking; treatment response; tumor; tumor microenvironment; tumor progression; uptake

ABSTRACT Head and neck cancer (HNSCC) afflicts mostly men. Female patients who develop HNSCC tend to be postmenopausal, with menopausal hormone therapy lowering their risk of developing the disease. We show that premenopausal female HNSCC patients have an improved response to therapy including immunotherapy (IO). Using clinical trial data, we show no difference in response to IO from testosterone. Similarly, our preclinical data show no IO response effect from gonadal castration. But 0.1 mg estradiol (E2) improves response in castrated male mice to that of female mice. Our overall objective is to understand how the premenopausal female hormone E2 contributes to progression and treatment resistance. Our data show that E2 had no effect on cancer cell viability, suggesting the tumor microenvironment (TME) as the site of action. E2 is known to play a major role in autoimmunity and inflammation, where it modulates dendritic cell (DC) and regulatory T cells (Treg) differentiation and function, and lymphatic vessel maturation. These immune cell populations regulate response to immunotherapy in HNSCC, but how they are affected by sex hormones remains largely uncharacterized. Our preclinical data also show that compared to male mice, females have increased DCs, less suppressive Tregs, and respond better to immunotherapeutic strategies with enhanced Teff function. Such response is removed with oophorectomy (OVX) but rescued with E2 of 0.1mg. OVX also changes the TME towards an immunosuppressive one with enhanced immunosuppressive Treg activity, similar to that of male mice. Finally, within the TME, OVX reduces the formation of high endothelial venules (HEVs) in the TME and draining lymph nodes (dLN), which enhance Teff cell priming and subsequent trafficking into the TME. We hypothesize that the sexually dimorphic response is driven by E2 acting via its receptors on Tregs and DCs to enhance antigen presentation and increase Teff activation, thus selectively determining response to immunotherapy. We also hypothesize that E2 will increase homing and trafficking of antigen specific T cells by enhancing HEV formation and maturation. In Aim1, we will interrogate the regulatory mechanisms of E2's effects on the DC biology by using genetically engineered mouse models (GEMMS), in vitro assays, and proteomic analysis to examine receptor subtypes and molecular mediators of DC differentiation, maturation, and antigen presentation. Aim 2 will dissect mechanisms of how E2's effect on Tregs can drive response and selection of sex-specific immunotherapies. We will test this with GEMMS, mouse chimeras, and pharmacological manipulation in in vivo and in vitro co-culture experiments, to identify signaling and downstream effectors. Focusing on T cell trafficking in Aim 3, we will examine E2's effect on HEV formation, and T cell egress from the dLN to the TME and systemic circulation using a combination of flow and immunofluorescence techniques in GEMMs and in vitro systems. These studies will collectively elucidate the basic mechanisms for advancing sex- directed novel biomarkers and therapeutics in HNSCC.

摘要 头颈癌（HNSCC）主要困扰男性。发展为HNSCC的女性患者往往 绝经后，绝经激素治疗降低了他们患上这种疾病的风险。我们证明了 绝经前女性HNSCC患者对包括免疫疗法（IO）在内的疗法具有改善的应答。 使用临床试验数据，我们显示对IO的反应与睾酮没有差异。同样，我们的临床前数据 显示性腺去势无IO反应效应。但0.1 mg雌二醇（E2）可改善去势大鼠的反应， 雄性小鼠与雌性小鼠相比。我们的总体目标是了解绝经前女性激素 E2有助于疾病进展和治疗抵抗。我们的数据表明，E2对癌细胞没有影响， 生存力，表明肿瘤微环境（TME）作为作用部位。众所周知，E2在以下方面发挥着重要作用： 自身免疫和炎症，其中它调节树突状细胞（DC）和调节性T细胞（Treg） 分化和功能以及淋巴管成熟。这些免疫细胞群调节反应 HNSCC的免疫治疗，但它们如何受到性激素的影响仍然在很大程度上没有特征。我们 临床前数据还显示，与雄性小鼠相比，雌性小鼠具有增加的DC，较少的抑制性T细胞， 并且对具有增强的Teff功能的免疫策略响应更好。这样的回应被删除 行卵巢切除术（OVX），但用0.1mg E2挽救。OVX还将TME改变为 具有增强的免疫抑制性Treg活性的免疫抑制性小鼠，类似于雄性小鼠。最后， 在TME内，OVX减少了TME和引流淋巴中高内皮微静脉（HEV）的形成 淋巴结（dLN），其增强Teff细胞引发和随后运输到TME中。我们假设 性二态反应由E2通过其受体作用于T细胞和DC来驱动， 抗原呈递并增加Teff活化，从而选择性地决定对 免疫疗法我们还假设E2会增加抗原特异性T细胞的归巢和运输， 通过增强戊型肝炎病毒的形成和成熟来增强细胞。在目标1中，我们将询问监管机制 通过使用基因工程小鼠模型（GEMMS）、体外试验和 蛋白质组学分析以检查DC分化、成熟和分化的受体亚型和分子介导物， 抗原呈递目标2将剖析E2对TTRY的影响如何驱动反应的机制， 选择性别特异性免疫疗法。我们将用GEMMS、小鼠嵌合体和药理学方法来测试这一点。 在体内和体外共培养实验中的操作，以鉴定信号传导和下游效应物。 关注目标3中的T细胞运输，我们将研究E2对HEV形成的影响，以及T细胞从目标3中的排出。 使用流动和免疫荧光技术的组合， GEMM和体外系统。这些研究将共同阐明促进性的基本机制- 指导HNSCC的新生物标志物和治疗。