Intratumoral immunotherapy to enhance T cell infiltration and augment immune checkpoint blockade responses across molecular subtypes of breast cancer

肿瘤内免疫疗法可增强 T 细胞浸润并增强乳腺癌分子亚型的免疫检查点阻断反应

• 批准号: 10689654
• 负责人: ERIKA J CROSBY
• 金额: $ 11.76万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10689654
• 关键词: Address; Antitumor Response; BRCA1 gene; Biological Markers; Breast Cancer Model; Breast Cancer Patient; Breast Cancer Treatment; CD8-Positive T-Lymphocytes; CRISPR screen; CXCL9 gene; CXCR3 gene; Cells; Clinical; Clinical Data; Clinical Research; Combination immunotherapy; Data; Data Set; Disease; ERBB2 gene; Electroporation; Electroporation Therapy; Engineering; Funding; Gene Targeting; Generations; Genes; Goals; Grant; Half-Life; Immune; Immune checkpoint inhibitor; Immunologics; Immunotherapy; Individual; Infiltration; Injections; Interleukin-12; K22 Award; Lesion; Licensing; Ligands; Mammary Neoplasms; Modeling; Molecular; Monitor; Oncogenes; Oncogenic; Outcome; PD-1/PD-L1; PTEN gene; Paclitaxel; Pathway interactions; Patients; Plasmids; Productivity; Progression-Free Survivals; Proteins; Publishing; Recombinant Interleukin-12; Recurrence; Research; Research Design; Resources; Role; Signal Transduction; Site; T cell infiltration; T-Lymphocyte; T-Lymphocyte Epitopes; TP53 gene; Testing; Time; Toxic effect; Transgenic Organisms; Translating; Treatment Protocols; Up-Regulation; Work; adaptive immune response; anti-PD-L1; anti-PD-L1 therapy; anti-tumor immune response; cancer infiltrating T cells; cancer type; chemokine; design; disorder control; follow-up; genetic signature; genomic tools; immune activation; immune cell infiltrate; immune checkpoint; immune checkpoint blockade; immunogenic; improved; improved outcome; individualized medicine; insight; knock-down; malignant breast neoplasm; migration; molecular subtypes; mouse model; phase 2 study; pre-clinical; prognostic; programs; receptor; response; single-cell RNA sequencing; survival outcome; systemic toxicity; trafficking; transcriptome sequencing; triple-negative invasive breast carcinoma; tumor; tumor microenvironment

1 Abstract 2 Breast cancer (BC) encompasses multiple diseases made up of different molecular subtypes that are 3 characterized by distinct oncogenic drivers and unique treatment regimens. Despite these differences, across all 4 subtypes, individuals with advanced, recurrent, or metastatic disease still have limited treatment options and 5 poor overall survival outcomes. Immunotherapies offer an opportunity to treat patients regardless of molecular 6 subtypes. This proposal leverages intratumoral (IT) immunotherapy as an alternative to ‘license’ treated lesions 7 to yield productive ratios of T cells to suppressive immune subsets while amplifying immune checkpoint axes 8 and ultimately increasing sensitivity to immune checkpoint inhibitors (ICI). Using a model of TNBC and single 9 cell RNA sequencing, we demonstrate that IT plasmid IL-12 (pIL-12) can convert poorly immunogenic/low 10 TIL tumors into highly inflamed, immunologically active lesions through the coordinated upregulation of the 11 CXCR3 axis in infiltrating immune cells that impacts the migration, differentiation, and activation of both 12 innate and adaptive immune cells. This CXCR3 signature was also significantly enhanced in patients that had 13 an increase in CD8 T cell infiltration into treated tumors post IT pIL-12 therapy and prognostic of improved 14 overall survival. We hypothesize that targeting the CXCR3 axis IT will enhance TILs and convert patients into 15 ICI responders across all molecular subtypes of BC. The proposed work will leverage this preliminary data in 16 the following 3 aims: 1.) Demonstrate increased infiltration of tumor-specific T cells following IT pIL-12-EP 17 treatment and validate the induction of a CXCR3 or trafficking-associated gene signature in ICI responders 2.) 18 Evaluate the role of trafficking-associated proteins in enhancing responsiveness to anti-PDL1 in TNBC using a 19 CRISPR based screen; 3.) Assess IT injection of plasmid CXCL9 to determine if direct targeting of the CXCR3 20 axis is sufficient to enhance T cell infiltration. Dr. Crosby’s long-term goal is to build a research program that 21 contributes to an understanding of immune cell infiltration into tumors to better design, combine, and predict 22 responses to immunotherapies, with a specific focus on BC. A critical impediment to these types of studies is 23 the lack of oncogene-driven, spontaneous BC tumor models which hampers the translational applicability of 24 many pre-clinical findings. Key resources for Dr. Crosby’s independent research are the spontaneous HER2- 25 driven and p53/BRCA1/PTEN-driven TNBC models that she has created, published, and will uniquely possess 26 to perform these and many other studies. Addressing the basic question of how to enhance T cell infiltration 27 into tumors has significant implications for changing the paradigm of treatment for BC patients, particularly 28 using an intratumoral plasmid approach that is easily altered to follow up on newly identified targets. The 29 funding provided by this K22 award will protect Dr. Crosby’s research time to develop and publish these 30 foundational, studies which will support subsequent R01 grant submissions.

1 摘要 2 乳腺癌 (BC) 涵盖由不同分子亚型组成的多种疾病，这些疾病 3 尽管存在这些差异，但仍具有不同的致癌驱动因素和独特的治疗方案。 4 种亚型，患有晚期、复发或转移性疾病的个体的治疗选择仍然有限，并且 5 总体生存结果较差的免疫疗法为治疗患者提供了机会，无论分子水平如何。 该提案利用瘤内 (IT) 免疫疗法作为“许可”治疗病变的替代疗法。 7 产生 T 细胞与抑制性免疫子集的生产率，同时放大免疫检查点轴 8 使用 TNBC 和单一模型最终提高对免疫检查点抑制剂 (ICI) 的敏感性。 9细胞RNA测序，我们证明IT质粒IL-12（pIL-12）可以将免疫原性差/低 10 TIL 肿瘤通过协调上调 11 浸润免疫细胞中的 CXCR3 轴影响两者的迁移、分化和激活 12 先天性和适应性免疫细胞的这种 CXCR3 特征在患有此病的患者中也显着增强。 13 IT pIL-12 治疗后，CD8 T 细胞浸润到治疗的肿瘤中有所增加，并且预后得到改善 14 总生存率我们勇敢地认为，针对 CXCR3 轴 IT 将增强 TIL 并将患者转化为 15 BC 所有分子亚型的 ICI 反应者拟议的工作将利用这些初步数据。 16 以下 3 个目标：1.) 证明 IT pIL-12-EP 后肿瘤特异性 T 细胞的浸润增加 17 治疗并验证 ICI 应答者中 CXCR3 或贩运相关基因特征的诱导 2.) 18 使用 TNBC 评估贩运相关蛋白在增强抗 PDL1 反应性方面的作用 19 基于 CRISPR 的筛选；3.) 评估质粒 CXCL9 的 IT 注射以确定是否直接靶向 CXCR3 20 轴足以增强 T 细胞浸润。 Crosby 博士的长期目标是建立一个研究计划， 21 有助于了解免疫细胞浸润到肿瘤中，以便更好地设计、组合和预测 22 对免疫疗法的反应，特别关注 BC。此类研究的一个关键障碍是。 23 缺乏癌基因驱动的自发性 BC 肿瘤模型，这阻碍了 24 Crosby 博士独立研究的许多临床前发现的关键资源是自发的 HER2-。 她创建、发布并将唯一拥有 25 个驱动和 p53/BRCA1/PTEN 驱动的 TNBC 模型 26 进行这些研究和许多其他研究，解决如何增强 T 细胞浸润的基本问题。 27 转化为肿瘤对于改变 BC 患者的治疗模式具有重大意义，特别是 28 使用肿瘤内质粒方法，可以轻松改变以跟踪新确定的靶标。 29 该 K22 奖项提供的资金将保护 Crosby 博士开发和发表这些成果的研究时间 30 项基础研究将支持后续 R01 拨款申请。