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.

项目摘要 像Sjnamegren的综合症（SJS）一样，自身免疫性SICCA可以发挥作用，或者作为免疫相关的敏锐 免疫粘液抑制剂（ICI）治疗癌症后不良事件（IRAE）。大约 每年有500,000名接受ICIS治疗的患者，估计有3-24％的患者出现新发明的SICCA症状；这些 随着FDA批准ICI使用扩展，数字将增加。我们定义ICI诱导的自身免疫性Sicca（ICIA- SICCA）作为10-25％的ICI-SICCA患者，患有可证明的B细胞自身免疫性，如RO52所示 或RO60自身抗体。需要新的，特定的生物标志物来预测谁将开发哪种类型的伊拉斯。 ICIA-SICCA中的“人类实验”可以比较RO自身免疫之前和之后的B细胞 与SJS“自然实验”相反，发展的主动性不是 已知。 B细胞克隆通过T细胞选择和B细胞受体（BCR）亲和力成熟而产生，但它是 尚不清楚这种高度焦点的B细胞反应是由ICI中的ICIA使用引起的，或者是哪个功能B 细胞子集（例如，内存）扩展。为了填补知识中的这些空白，我们建立了精心策划的生物库 在ICI治疗前从患者那里收集的外周血样本和ICIA-SICCA在发育中 与M.D. Doug Johnson，M.S.C.I.，Vanderbilt黑色素瘤研究计划主管和IRAE合作 专家。高通量人类杂交瘤技术将用于AIM 1来识别分子特征 由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的自动反应性B细胞扩张中发挥作用。外周血膨胀 在SJS患者中观察到CD21LO B细胞（一个自动反应性易发的子集），并预测ICI治疗的IRAE 患者。因此，我们将通过比较单个单一的单一和其他B细胞子集的扩展来调查单个B细胞子集的扩展 细胞曲目（BCRSEQ），表型（CITESEQ）和icia-Sicca中的转录组（RNASEQ）B细胞特征 ICI治疗前的患者并遵循ICIA-SICCA发育。我们将使用这些数据来识别 ICIA-SICCA中扩展的B细胞克隆和子集的标志。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 B细胞的功能能力和发育起源，识别ICIA中与SJS相关的自身抗原 Sicca。识别ICIA-SICCA中其他自身抗原的自动反应性B细胞将由无偏见识别 AIM 2中的方法。这些研究将发现可以使用的特定序列和表型生物标志物 将来可以预测即将来临的ICIA-SICCA并评估ICIA-SICCA的免疫抑制效率。人类 RO52和RO60单克隆抗体，自动反应性BCR图案数据库和平行 我们将生成的BCRSEQ/CITESEQ/RNASEQ数据将公开用于支持IRAE和 赞助自身免疫性疾病研究。我们将创建的实验和分析蓝图将设置 ICI治疗后，未来研究研究其他伊拉斯的起源的阶段。