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: 10283773
• 负责人: ERIKA J CROSBY
• 金额: $ 16.15万
• 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/10283773
• 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; Engineering; Foundations; 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; Progression-Free Survivals; Proteins; Publishing; Receptor Signaling; Recombinant Interleukin-12; Recurrence; Research; Research Design; Resources; Role; Site; T-Lymphocyte; T-Lymphocyte Epitopes; TP53 gene; Testing; Time; Toxic effect; Transgenic Organisms; Translating; Treatment Protocols; Tumor-infiltrating immune cells; Up-Regulation; Work; adaptive immune response; anti-PD-L1; anti-tumor immune response; base; cancer type; chemokine; design; disorder control; follow-up; genetic signature; genomic tools; immune activation; 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; 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）包括由不同的分子亚型组成的多种疾病 3特征是不同的致癌驱动因素和独特的治疗方案。尽管存在这些差异， 4个亚型，患有晚期，经常性或转移性疾病的个体仍有有限的治疗选择，并且 5个整体生存结果差。免疫疗法提供了治疗患者的机会 6个亚型。该建议利用肿瘤内（IT）免疫疗法作为“许可”治疗腿的替代方案 7在放大免疫传输检查点轴的同时，产生T细胞与抑制免疫物的生产比 8并最终提高对免疫抑制剂（ICI）的敏感性。使用TNBC和单个模型 9细胞RNA测序，我们证明了质粒IL-12（PIL-12）可以转化不良的免疫原性/低 10通过协调的上调到高度发炎的免疫主动病变中的10个肿瘤 11 CXCR3轴在浸润的免疫电池中影响迁移，分化和激活 12个先天和适应性免疫小球。对于患者，此CXCR3签名也显着增强 13 CD8 T细胞在PIL-12疗法后浸润到治疗的肿瘤中的预后改善 14总体生存。我们假设靶向CXCR3轴它将增强tils并将患者转化为 BC的所有分子亚型中的15个ICI响应者。拟议的工作将利用此初步数据 16以下3个目的：1。）表明肿瘤特异性T细胞的浸润增加了IT PIL-12-EP 17治疗并验证ICI响应者中CXCR3或与人口相关的基因签名的诱导2.） 18使用A增强对抗PDL1的反应性，评估了使用A的作用 19基于CRISPR的屏幕； 3.）评估IT注入质粒CXCL9，以确定CXCR3的直接靶向 20轴足以增强T细胞浸润。克罗斯比博士的长期目标是建立一个研究计划 21有助于理解免疫细胞对肿瘤的浸润，以更好地设计，结合和预测 22对免疫疗法的反应，特别关注BC。这类研究的关键障碍是 23缺乏致癌基因驱动的赞助BC肿瘤模型，这阻碍了翻译的适用性 24许多临床前发现。 Crosby博士独立研究的关键资源是赞助的HER2- 她创建，出版并将拥有25驱动器和P53/BRCA1/PTEN驱动的TNBC模型 26进行这些和许多其他研究。解决如何增强T细胞浸润的基本问题 27进入肿瘤对改变卑诗省患者的治疗范例具有重要意义 28使用肿瘤内质粒方法很容易改变以跟进新确定的目标。这 29个K22奖提供的资金将保护克罗斯比博士的研究时间来开发和发布这些 30基础，将支持随后的R01赠款提交的研究。