Mechanisms and therapeutic targeting of osteoimmune functions of RANKL in breast cancer

RANKL在乳腺癌中的骨免疫功能的机制和治疗靶点

基本信息

批准号：
10586000
负责人：
Selvarangan Ponnazhagan
金额：
$ 44.67万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10586000
关键词：
Address Adopted Affinity Antibodies Apoptosis Autocrine Communication Binding Bone Diseases Breast Cancer Cell Breast Cancer Model Breast Cancer Risk Factor Breast cancer metastasis CD8-Positive T-Lymphocytes Cell Therapy Cell physiology Cells Clinic Combination Drug Therapy Combined Modality Therapy Disease Disease Progression Double-Blind Method Engineering Environment G-Protein-Coupled Receptors Goals Homeostasis Human Human Pathology Immune Immune Targeting Immune system Immunocompetent Immunocompromised Host Immunosuppression Induction of Apoptosis Injections International Leucine-Rich Repeat Ligand Binding Ligands Link Macrophage Malignant Bone Neoplasm Mediator Metastatic Neoplasm to the Bone Metastatic breast cancer Molecular Monoclonal Antibodies Monoclonal Antibody Therapy Mus Myeloid Cells Myeloid-derived suppressor cells Neoplasm Metastasis Nuclear Osteoclasts Osteolysis Outcome Study Pathology Pathway interactions Patients Phase Placebo Control Play Primary Neoplasm Production Protein Engineering Proteins Publishing Randomized Recurrent disease Refractory Resistance Role Signal Transduction Skeletal system Structure Systemic Therapy TNF-related apoptosis-inducing ligand TRANCE protein Testing Therapeutic Therapeutic Effect Tumor Promotion Tumor necrosis factor receptor 11b Variant antagonist bone chemotherapy cytokine design effector T cell exhaustion genetically modified cells high risk immune checkpoint blockade immune resistance improved in vivo in vivo Model malignant breast neoplasm monocyte mouse model neoplastic cell new combination therapies novel paracrine phase 3 study polarized cell programmed cell death ligand 1 programmed cell death protein 1 protein structure receptor receptor function skeletal skeletal-related events spatiotemporal targeted treatment taxane therapeutic target tumor tumor growth tumor microenvironment

项目摘要

Complexities in treating breast cancer (BCa) with bone metastasis are aggravated by a vicious protumorigenic pathology involving a shift in skeletal homeostasis towards aggressive osteoclast activity and polarization of myeloid cells, favoring M2 macrophage (MФ) and myeloid-derived suppressor cell (MDSC) accumulation as key mediators of immunosuppression. In addition to the tumor cells, protumorigenic myeloid cells contribute to the cascade by expressing checkpoint ligands, blunting antitumor functions of effector T cells. Hence, a better understanding of key signaling mechanisms that alter skeletal and immune homeostasis towards protumorigenic functions will enable the designing of new combination therapies targeting this biphasic effect. In this pursuit, we have identified that in addition to robust activation of osteoclast precursors, receptor activator of nuclear factor kappa-Β ligand (RANKL) plays an important role as an osteoimmune link in MФ polarization and programmed death-ligand 1 (PD-L1) expression. We identified that elevated RANKL from BCa cells induce paracrine effects on differentiation of monocytes to immunosuppressive M2 MФ in a spatiotemporal manner. Preliminary studies presented in this application indicate that whereas RANKL canonical autocrine signaling via RANK activates a feed-forward loop in BCa cells, non-canonical RANKL signaling enhances PD-L1 expression in M2 MФ and MDSCs via the leucine-rich repeat containing G-protein coupled receptor (Lgr4). Based on our published and preliminary findings, the overarching goal of this proposal is to expand our understanding on the pleiotropic mechanisms of RANKL in BCa immunosuppression and bone damage, and to test the potential of combining a novel osteoprotegerin (OPG) cell therapy without interfering in TNF-related apoptosis-inducing ligand (TRAIL) function, with checkpoint blockade and chemotherapies, to reverse tumor- associated pathology in the immune and skeletal systems. We recently adopted a protein structure-based engineering approach and identified a critical domain on OPG for TRAIL binding and successfully developed and validated in vivo an OPG variant (OPGY49R) that retains RANKL binding, but lacks TRAIL binding. Preliminary studies, directly comparing a cell-based, single-application OPGY49R treatment with multiple applications of a neutralizing RANKL mAb therapy indicated systemically stable levels of OPGY49R from a single injection and a significant decrease in CD8+ T cell exhaustion, compared to RANKL mAb treatment. More importantly, OPGY49R greatly decreased metastasis of primary tumors in vivo, demonstrating its potential advantage over the RANKL mAb, denosumab, which failed to delay bone metastasis or disease recurrence in patients with high-risk early- stage BCa in a recent international double-blinded randomized placebo-controlled, phase 3 study (D-CARE). This proposal will test this novel, biologically driven combination therapy approach by using immunocompetent mouse models of BCa, as applicable to both pre-metastatic and metastatic disease.

恶性促肿瘤因素加剧了治疗伴有骨转移的乳腺癌 (BCa) 的复杂性涉及骨骼稳态向侵袭性破骨细胞活性和极化的转变的病理学骨髓细胞，有利于 M2 巨噬细胞 (MФ) 和骨髓源性抑制细胞 (MDSC) 积累是关键除了肿瘤细胞外，促肿瘤骨髓细胞也有助于免疫抑制。通过表达检查点配体，削弱效应 T 细胞的抗肿瘤功能，从而产生更好的级联反应。了解改变骨骼和免疫稳态以促进肿瘤发生的关键信号机制功能将使针对这种双相效应的新组合疗法的设计成为可能。已经发现，除了破骨细胞前体的强烈激活之外，核因子的受体激活剂 kappa-β 配体 (RANKL) 作为骨免疫环节在 MФ 极化和程序化中发挥着重要作用我们发现 BCa 细胞的 RANKL 表达升高会诱导旁分泌效应。单核细胞向免疫抑制 M2 MФ 的时空分化的初步研究。本申请中提出的 RANKL 规范自分泌信号通过 RANK 激活 BCa 细胞中的前馈回路，非经典 RANKL 信号传导增强 M2 MФ 和 MФ 中的 PD-L1 表达 MDSC 通过含有 G 蛋白偶联受体 (Lgr4) 的富含亮氨酸重复序列。根据我们已发表的初步调查结果，该提案的总体目标是扩大我们的了解 RANKL 在 BCa 免疫抑制和骨损伤中的多效机制，并测试结合新型骨保护素 (OPG) 细胞疗法而不干扰 TNF 相关的潜力凋亡诱导配体（TRAIL）功能，通过检查点阻断和化疗来逆转肿瘤我们最近采用了基于蛋白质结构的相关病理学。工程方法并确定了 OPG 上用于 TRAIL 结合的关键结构域并成功开发并在体内验证了保留 RANKL 结合但缺乏 TRAIL 结合的 OPG 变体 (OPGY49R)。研究，直接比较基于细胞的单次应用 OPGY49R 治疗与多次应用中和 RANKL mAb 疗法表明单次注射和一次注射后 OPGY49R 的系统水平稳定。与 RANKL mAb 治疗相比，CD8+ T 细胞耗竭显着减少，更重要的是，OPGY49R。大大减少了体内原发肿瘤的转移，证明了其相对于 RANKL 的潜在优势单克隆抗体、狄诺塞麦未能延缓高危早期患者的骨转移或疾病复发最近的一项国际双盲随机安慰剂对照 3 期研究 (D-CARE) 已达到 BCa 期。该提案将通过使用免疫活性来测试这种新颖的、生物驱动的联合治疗方法 BCa 小鼠模型，适用于转移前和转移性疾病。