Molecular Characterization of Autoreactive B Cells in Immune Checkpoint Inhibitor-Induced Autoimmune Sicca

免疫检查点抑制剂诱导的自身免疫性干燥症中自身反应性 B 细胞的分子特征

• 批准号: 10287167
• 负责人: Rachel H Bonami
• 金额: $ 8.65万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10287167
• 关键词: Activities of Daily Living; Acute; Affinity; Antigens; Autoantibodies; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmunity; B lymphocyte immortalization; B-Cell Antigen Receptor; B-Cell Receptor Binding; B-Lymphocyte Subsets; B-Lymphocytes; B-cell receptor repertoire sequencing; Binding; Biological Markers; Blood specimen; Cancer Patient; Cell surface; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Clinical; Clonal Evolution; Clone Cells; Collaborations; Communities; Computerized Medical Record; Data; Data Set; Databases; Development; Diagnostic; Disease; Doctor of Medicine; Exhibits; Female; Frequencies; Future; Genes; Genetic Transcription; Human; Hybridomas; Immune Tolerance; Immune checkpoint inhibitor; Immune system; Immunosuppression; Individual; Insulin-Dependent Diabetes Mellitus; Investigation; Knowledge; Letters; Memory; Molecular; Monoclonal Antibodies; Mutate; Mutation; Nature; Oncologist; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Phylogenetic Analysis; Plasmablast; Play; Prednisone; Production; Receptor Gene; Research; Risk; Role; Shapes; Sjogren' s Syndrome; Symptoms; Syndrome; T cell response; T-Lymphocyte; Technology; Trees; Work; autoimmune pathogenesis; autoreactive B cell; autoreactivity; beta diversity; biobank; cancer therapy; checkpoint therapy; design; experimental analysis; experimental study; immune-related adverse events; improved; insight; man; melanoma; new technology; peripheral blood; phenotypic biomarker; phenotypic data; programs; recruit; response; rituximab; searchable database; specific biomarkers; tool; transcriptome sequencing; transcriptomics; tumor

PROJECT SUMMARY Autoimmune sicca can arise spontaneously, as in Sjӧgren’s syndrome (SjS), or acutely, as an immune-related adverse event (irAE) following immune checkpoint inhibitor (ICI) therapy for cancer. Of the approximately 500,000 patients treated with ICIs annually, an estimated 3-24% develop new-onset sicca symptoms; these numbers will increase as FDA approvals for ICI use broaden. We define ICI-induced autoimmune sicca (ICIA- sicca) as the 10-25% of ICI-sicca patients who develop demonstrable B cell autoimmunity, as shown by Ro52 or Ro60 autoantibodies. New, specific biomarkers are needed to predict who will develop which types of irAEs. The “experiment of man” in ICIA-sicca enables comparison of B cells before and after Ro autoimmunity develops, in contrast to the SjS “experiment of nature”, in which the initiation point of autoimmunity is not known. B cell clones arise via T cell selection and B cell receptor (BCR) affinity maturation in SjS, but it is unknown whether such highly focused B cell responses result from ICI use in ICIA-sicca, or which functional B cell subsets (e.g. memory) are expanded. To fill these gaps in knowledge, we built a carefully curated biobank of peripheral blood samples collected from patients before ICI therapy and following ICIA-sicca development in collaboration with Doug Johnson, M.D., M.S.C.I., Vanderbilt Melanoma Research Program Director and irAE expert. High-throughput human hybridoma technology will be used in Aim 1 to identify the molecular features of BCRs expressed by Ro52 and Ro60-binding B cells. We will discern the role that mutation and selection plays in autoreactive B cell expansion that precedes autoantibody production. Peripheral blood expansion of CD21lo B cells (an autoreactive-prone subset) is observed in SjS patients and predicts irAEs in ICI-treated patients. We will therefore investigate expansion of this and other B cell subsets in Aim 2 by comparing single- cell repertoire (BCRseq), phenotypic (CITEseq), and transcriptomic (RNAseq) B cell signatures in ICIA-sicca patients before ICI treatment and following ICIA-sicca development. We will use these data to identify hallmarks of expanded B cell clones and subsets in ICIA-sicca. We will further integrate Aim 1 and Aim 2 data to determine Ro52/Ro60-specific V gene identity, clonal relatedness, and phenotypic information to infer the functional capacity and developmental origins of B cells that recognize SjS-associated autoantigens in ICIA- sicca. Autoreactive B cells that recognize other autoantigens in ICIA-sicca will be identified by the unbiased approach in Aim 2. These studies will uncover specific sequence and phenotypic biomarkers that can be used in the future to predict impending ICIA-sicca and assess immunosuppressive efficacy in ICIA-sicca. Human Ro52 and Ro60 monoclonal antibodies, the autoreactive BCR motif database, and the parallel BCRseq/CITEseq/RNAseq data we will generate will be made publicly available to support irAE and spontaneous autoimmune disease research. The experimental and analysis blueprints we will create will set the stage for future studies to investigate the origins of other irAEs following ICI treatment.

项目概要 自身免疫性干燥可以自发出现，如干燥综合征 (SjS)，也可以急性出现，如免疫相关疾病 癌症免疫检查点抑制剂（ICI）治疗后的不良事件（irAE）。其中大约 每年有 500,000 名接受 ICI 治疗的患者，估计有 3-24% 出现新发干燥症状；这些 随着 FDA 对 ICI 使用范围的批准扩大，数量将会增加。我们定义 ICI 诱导的自身免疫性干燥 (ICIA- 干燥症），如 Ro52 所示，10-25% 的 ICI-干燥症患者出现明显的 B 细胞自身免疫 或 Ro60 自身抗体。需要新的、特定的生物标志物来预测谁将出现哪种类型的 irAE。 ICIA-sicca 中的“人体实验”可以比较 Ro 自身免疫前后的 B 细胞 与 SjS“自然实验”相反，其中自身免疫的起始点不是 已知。 B 细胞克隆是通过 SjS 中的 T 细胞选择和 B 细胞受体 (BCR) 亲和力成熟而产生的，但它是 尚不清楚这种高度集中的 B 细胞反应是否是由于 ICIA-sicca 中使用 ICI 引起的，或者是哪种功能性 B 细胞反应所致。 细胞子集（例如记忆）得到扩展。为了填补这些知识空白，我们建立了一个精心策划的生物库 在 ICI 治疗前和 ICIA-干燥发展后从患者收集的外周血样本 与 Doug Johnson 医学博士、MSCI、范德比尔特黑色素瘤研究项目主任和 irAE 合作 专家。目标1将使用高通量人类杂交瘤技术来鉴定分子特征 Ro52 和 Ro60 结合 B 细胞表达的 BCR。我们将辨别突变和选择的作用 在自身抗体产生之前参与自身反应性 B 细胞扩增。外周血扩张 在 SjS 患者中观察到 CD21lo B 细胞（易发生自身反应的子集），并预测 ICI 治疗中的 irAE 患者。因此，我们将通过比较单细胞来研究目标 2 中该 B 细胞亚群和其他 B 细胞亚群的扩展。 ICIA-sicca 中的细胞库 (BCRseq)、表型 (CITEseq) 和转录组 (RNAseq) B 细胞特征 ICI 治疗前和 ICIA-干燥发展后的患者。我们将使用这些数据来识别 ICIA-sicca 中扩增的 B 细胞克隆和亚群的标志。我们将进一步整合Aim 1和Aim 2数据 确定 Ro52/Ro60 特异性 V 基因身份、克隆相关性和表型信息，以推断 ICIA-中识别 SjS 相关自身抗原的 B 细胞的功能能力和发育起源 干燥。识别 ICIA-sicca 中其他自身抗原的自身反应性 B 细胞将被公正的鉴定 目标 2 中的方法。这些研究将揭示可使用的特定序列和表型生物标志物 未来预测即将发生的 ICIA-sicca 并评估 ICIA-sicca 的免疫抑制功效。人类 Ro52 和 Ro60 单克隆抗体、自身反应 BCR 基序数据库以及并行数据库 我们将生成的 BCRseq/CITEseq/RNAseq 数据将公开以支持 irAE 和 自发性自身免疫性疾病研究。我们将创建的实验和分析蓝图将设定 这是未来研究 ICI 治疗后其他 irAE 起源的研究阶段。