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Anti-programmed death-1 antibody (αPD-1 Ab) as a single agent for treating human hepatocellular cancer (HCC) was withdrawn from the US market on July 26, 2021, because a multi-center phase III study did not demonstrate its efficacy in improving patient survival over controls.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.

摘要 抗程序性死亡-1抗体(αPD-1 Ab)作为单药治疗人肝细胞癌的研究 于2021年7月26日从美国市场撤出，因为一项多中心III期研究没有证明 与对照组相比，它在提高患者存活率方面的有效性。因此，迫切需要确定和确定关键的目标 细胞和分子调节器，以设计新的免疫检查点治疗(ICT)策略，以对抗肝癌。一个 建立了独特的、原位的和临床相关的小鼠肝癌模型，该模型模拟了肝癌的起源和发展 它反映了人类疾病典型的肿瘤生物学、免疫学和组织学特征。在……里面 在这个模型中，SV40T抗原(Tag)作为可追踪的肿瘤特异性抗原(TSA)仅在肿瘤中表达， 在肿瘤的起始、进展和治疗反应期间进行TSA免疫研究。使用这个模型， 人们开发了几种基于免疫的肝癌治疗策略，其中一种新的基于微生物的策略是 最近证实，显著提高了αPD-1抗体对肝癌的治疗效果。具体来说， 类杆菌thetaiotaomicron(B.th)是类杆菌属的一员，其富含CpG的核酸 作为TLR9激动剂，被确定为显著促进αPD-1抗体治疗的微生物调节剂 对肝细胞癌的疗效。这一令人兴奋的发现导致了对潜在的细胞和分子机制的研究。单人- 细胞RNA测序(scRNA-seq)显示，肝癌生长上调了肿瘤组织中的Kruppel样因子-2(KLF2)。 辅助性巨噬细胞(TAM)和B.th激活的效应T细胞和改进的αPD-1 对肝癌的治疗作用与其降低TAMs中KLF2的表达有关。此外， KLF2基因敲除的巨噬细胞(MΦS)过继细胞转移可诱导TSA免疫 显著抑制肝癌生长。相反，KLF2在MΦS中的过表达(OE)抑制了B.TH/αPD-1- 诱导对肝癌的治疗性抑制。这些令人信服的结果突显了KLF2作为一个关键的调节器 微生物对肝癌发生的影响及B.Th/αPD-1免疫治疗调节M-ΦS的进一步研究 提示KLF2通过调控TLR9的MΦ表达和信号调节蛋白α(Sirpα)来调节MΦ 肿瘤吞噬和免疫调节功能。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M-α重振肝癌中的ΦPD-1AbICT 通过KLF2调控TLR9和Sirpα的表达。在目标1中，KLF2指导的MΦS作为细胞基础介导 肝癌的发病机制和免疫耐受的研究将朝着开发新的治疗方法的方向发展 通过将KLF2-KO MΦS与 αPD-1抗体在目标2中，分子机制和调节因子 B.Th/αPD-1通过调控TLR9和Sirpα的表达抑制KLF2对M-Sirp的重新编程。这个 这项研究产生的知识不仅将确定未被认识的内源性调节因子在 在肝癌中规划MΦ，也说明这些因素是触发MΦS和铅的有效靶点 为改善肝细胞癌的信息和通信技术。