Data_Sheet_1_Examining Peripheral and Tumor Cellular Immunome in Patients With Cancer.PDF

Immunotherapies are rapidly being integrated into standard of care (SOC) therapy in conjunction with surgery, chemotherapy, and radiotherapy for many cancers and a large number of clinical studies continue to explore immunotherapy alone and as part of combination therapies in patients with cancer. It is evident that clinical effectiveness of immunotherapy is limited to a subset of patients and improving immunotherapy related outcomes remains a major scientific and clinical effort. Understanding the immune cell subset phenotype and activation/functional status (cellular immunome) prior to and post therapy is therefore critical to develop biomarkers that (1) will predict if a patient will respond to immunotherapy and (2) are a result of immunotherapy. In this study, we investigated local (tumor) and peripheral (blood) cellular immunome of patients with melanoma, breast cancer, and brain cancer using a rapid and reliable standardized, multiparameter flow cytometry assay. We used this approach to monitor changes in the peripheral cellular immunome in women with breast cancer undergoing SOC therapy. Our analysis is unique because it is conducted using matched fresh tumor tissue and blood from patients in real-time, within 2–3 h of sample acquisition, and provides insight into the innate and adaptive immune cell profile in blood and tumor. Specific to blood, this approach involves no manipulation and evaluates all immune subsets such as T cells, B cells, natural killer (NK) cells, monocytes, dendritic cells (DCs), neutrophils, eosinophils, and basophils using 0.5 ml of blood. Analysis of the corresponding tumor provides much needed insight into the phenotype and activation status of immune cells, especially T and B cells, in the tumor microenvironment vs. the periphery. This analysis will be used to assess baseline and therapy-mediated changes in local and peripheral cellular immunome in patients with glioblastoma, breast cancer, and melanoma in planned immunotherapy clinical studies.

免疫治疗正快速融入多种癌症的标准治疗（standard of care, SOC）体系，常与手术、化疗及放疗联合应用；目前大量临床研究仍在探索单药免疫治疗，以及其作为联合治疗组分在癌症患者中的应用效果。显而易见，免疫治疗的临床有效性仅局限于部分患者群体，因此改善免疫治疗相关结局仍是当前科研与临床领域的核心攻关方向。因此，明确治疗前后免疫细胞亚群的表型与激活/功能状态（细胞免疫组，cellular immunome），对于开发两类生物标志物至关重要：其一可预测患者对免疫治疗的应答情况，其二可反映免疫治疗的作用效应。本研究采用快速可靠的标准化多参数流式细胞术检测方法，对黑色素瘤、乳腺癌及脑癌患者的局部（肿瘤组织）与外周（血液）细胞免疫组进行了分析。研究同时利用该方法，对接受标准治疗的乳腺癌女性患者的外周血细胞免疫组变化进行了监测。本分析的独特之处在于：我们采用配对的新鲜肿瘤组织与血液样本，在样本采集后2~3小时内完成实时检测，可全面解析血液与肿瘤组织中的先天免疫及适应性免疫细胞谱特征。针对血液样本，该方法无需额外处理，仅需0.5ml血液即可全面检测所有免疫细胞亚群，包括T细胞、B细胞、自然杀伤（natural killer, NK）细胞、单核细胞、树突状细胞（dendritic cells, DCs）、中性粒细胞、嗜酸性粒细胞及嗜碱性粒细胞。对配对肿瘤组织的分析，则可针对性解析肿瘤微环境中免疫细胞（尤其是T、B细胞）的表型与激活状态，并与外周免疫细胞特征形成对照。本分析方法将用于后续计划开展的免疫治疗临床研究中，评估胶质母细胞瘤、乳腺癌及黑色素瘤患者局部与外周细胞免疫组的基线水平及治疗诱导的变化。