Targeting KLF2 in macrophages to improve immune checkpoint therapy for hepatocellular cancer

靶向巨噬细胞中的 KLF2 以改善肝细胞癌的免疫检查点治疗

• 批准号: 10677187
• 负责人: Guangfu Li
• 金额: $ 56.75万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677187
• 关键词: 16S ribosomal RNA sequencing; Adoptive Cell Transfers; Agonist; Androgens; Antibiotic Therapy; Antibodies; Antibody Therapy; Antigens; BAY 54-9085; Bacteroides; Bacteroides thetaiotaomicron; C57BL/6 Mouse; CD8-Positive T-Lymphocytes; CTLA4 gene; Cancer Model; Carbon Tetrachloride; Cells; Combined Modality Therapy; Development; Disease; Growth; Hepatic; Hepatocarcinogenesis; Hepatocyte; Histology; Homeostasis; Human; Immune; Immune Tolerance; Immune response; Immunity; Immunization; Immunology; Immunotherapeutic agent; Immunotherapy; Inflammatory; Knowledge; Ligands; Liver; Liver Fibrosis; Macrophage; Malignant Neoplasms; Marketing; Mediating; Microbe; Modeling; Molecular; Mus; Myeloid-derived suppressor cells; Nucleic Acids; Oncogenic; PTPNS1 gene; Pathogenesis; Patients; Phagocytosis; Phenotype; Play; Population; Primary carcinoma of the liver cells; Progression-Free Survivals; Regulatory T-Lymphocyte; Resistance; Role; SHPS-1 protein; SV40 T Antigens; Shapes; Testing; Therapeutic; Therapeutic Effect; Therapeutic antibodies; Transgenic Mice; Treatment Efficacy; Tumor Antigens; Tumor Biology; Tumor Immunity; Tumor-associated macrophages; Unresectable; Up-Regulation; anti-PD-1; anti-PD-L1 antibodies; anti-PD1 antibodies; cancer immunotherapy; cancer initiation; checkpoint therapy; clinically relevant; design; exhaust; fecal transplantation; gut microbiota; human disease; immune activation; improved; in vivo; invention; knock-down; loss of function; melanoma; member; microbial; monocyte; novel; novel therapeutic intervention; overexpression; phase 3 study; programmed cell death ligand 1; programmed cell death protein 1; programs; single-cell RNA sequencing; transcription factor; treatment response; tumor; tumor growth; tumor initiation; tumor microbiota; tumor progression; tumorigenesis

ABSTRACT Anti-programmed death-1 antibody (αPD-1 Ab) as a single agent for treating human hepatocellular cancer (HCC) was withdrawn from the US market on July 26, 2021, because a multi-center phase III study did not demonstrate its efficacy in improving patient survival over controls. Thus, there is an urgent need to identify and target critical cellular and molecular regulators to design new immune checkpoint therapy (ICT) strategies against HCC. A unique, orthotopic, and clinically relevant murine HCC model was established that mimics HCC initiation and progression in humans and reflects the tumor biology, immunology, and histology typical of human disease. In this model, SV40 T antigen (TAg) is expressed solely in tumors as a trackable tumor-specific antigen (TSA), enabling TSA immunity study during tumor initiation, progression, and response to treatments. Using this model, several immune-based therapeutic strategies for HCC were developed and a novel microbe-based strategy was recently established that significantly improves the therapeutic efficacy of αPD-1 Ab for HCC. Specifically, Bacteroides thetaiotaomicron (B.th), one member of genus Bacteroides, with CpG-rich nucleic acid which functions as TLR9 agonist, was identified as a microbial regulator to significantly boost αPD-1 Ab therapeutic efficacy for HCC. This exciting finding led to studies of the underlying cellular and molecular mechanisms. Single- cell RNA sequencing (scRNA-seq) revealed that HCC growth upregulated Kruppel like factor-2 (KLF2) in tumor- associated macrophages (TAMs), and B.th addition activated effector CD8+ T cells and improved αPD-1 therapeutic effect against HCC, which was associated with reduced KLF2 expression in TAMs. Moreover, adoptive cell transfer (ACT) of macrophages (MΦs) with KLF2 knockdown (KD) enabled TSA immunization to significantly suppress HCC growth. Conversely, KLF2 overexpression (OE) in MΦs compromised B.th/αPD-1- induced therapeutic suppression of HCC. These compelling results highlight KLF2 as a key regulator mediating microbes’ impact on hepatocarcinogenesis and B.th/αPD-1 immunotherapy by modulating MΦs. Further studies indicate that KLF2 controls MΦ expression of TLR9 and signal-regulatory protein α (SIRPα) to regulate MΦ tumor phagocytosis and immune regulatory function. These findings generate the following hypothesis: B.th, with CpG-rich nucleic acid, reinvigorates αPD-1 Ab ICT in HCC by phenotypically and functionally programming MΦ via KLF2-controlled expression of TLR9 and SIRPα. In Aim 1, KLF2-directed MΦs as the cellular basis mediating HCC pathogenesis and immune tolerance will be studied toward the development of a new therapeutic approach by integrating KLF2-KO MΦs with αPD-1 Ab. In Aim 2, the molecular mechanism and regulators by which B.th/αPD-1 suppress KLF2 to reprogram MΦ by controlling TLR9 and SIRPα expression will be studied. The knowledge generated from this study will not only identify unrecognized endogenous regulators with a role in programming MΦ in HCC, but also make a case that these factors are effective targets to trigger MΦs and lead to improved ICT for HCC.

抽象的 抗程序性死亡 1 抗体 (αPD-1 Ab) 作为单一药物治疗人肝细胞癌 (HCC) 于2021年7月26日从美国市场撤出，因为多中心III期研究未证明 与对照相比，其在改善患者生存方面的功效因此，迫切需要识别和瞄准关键的目标。 细胞和分子调节剂设计针对 HCC A 的新免疫检查点疗法 (ICT) 策略。 建立了独特的、原位的、临床相关的小鼠 HCC 模型，模拟 HCC 的发生和发展 在人类中，反映了人类疾病典型的肿瘤生物学进展、免疫学和组织学。 在该模型中，SV40 T 抗原 (TAg) 仅在肿瘤中表达为可追踪的肿瘤特异性抗原 (TSA)， 使用该模型能够在肿瘤发生、进展和治疗反应期间进行 TSA 免疫研究， 开发了几种基于免疫的 HCC 治疗策略，并开发了一种新的基于微生物的策略 最近发现显着提高 αPD-1 Ab 对 HCC 的治疗效果。 Bacteroides thetaiotaomicron (B.th)，拟杆菌属成员之一，具有富含 CpG 的核酸， 作为 TLR9 激动剂，被确定为微生物调节剂，可显着增强 αPD-1 Ab 治疗效果 这一令人兴奋的发现引发了对潜在细胞和分子机制的研究。 细胞 RNA 测序 (scRNA-seq) 揭示，HCC 生长上调肿瘤中的 Kruppel 样因子 2 (KLF2) 相关巨噬细胞 (TAM) 和 B.th 附加激活效应 CD8+ T 细胞和改进的 αPD-1 对 HCC 的治疗作用，这与 TAM 中 KLF2 表达的减少有关。 巨噬细胞 (MΦs) 的过继细胞转移 (ACT) 与 KLF2 敲低 (KD) 使 TSA 免疫能够 显着抑制离线 HCC 生长，MΦ 中的 KLF2 过表达 (OE) 受损 B.th/αPD-1- 这些引人注目的结果凸显了 KLF2 作为介导的关键调节因子。 微生物通过调节 MΦs 对肝癌发生和 B.th/αPD-1 免疫治疗的影响。 表明KLF2控制TLR9和信号调节蛋白α（SIRPα）的MΦ表达来调节MΦ 这些发现产生了以下假设：B.th，具有。 富含 CpG 的核酸通过表型和功能编程 MΦ 重振 HCC 中的 αPD-1 Ab ICT 通过 KLF2 控制的 TLR9 和 SIRPα 表达在目标 1 中，KLF2 指导的 MΦ 作为介导的细胞基础。 将研究 HCC 发病机制和免疫耐受，以开发新的治疗方法 通过将 KLF2-KO MΦ 与 αPD-1 Ab 在目标 2 中，其分子机制和调节因子。 将研究 B.th/αPD-1 通过控制 TLR9 和 SIRPα 表达来抑制 KLF2 重编程 MΦ。 这项研究产生的知识不仅将识别出未识别的内源性调节因子 在 HCC 中规划 MΦ，同时也证明这些因素是触发 MΦ 并导致 改善 HCC 的 ICT。