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

• 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
• 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

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.

恶性促癌物加剧了乳腺癌骨转移治疗的复杂性 病理涉及骨骼动态平衡向侵袭性破骨细胞活动和极化的转变 髓系细胞，支持M2巨噬细胞(MФ)和髓系来源的抑制细胞(MDSC)聚集是关键 免疫抑制的介体。除了肿瘤细胞外，促肿瘤的髓系细胞也对 通过表达检查点配体而级联，钝化效应器T细胞的抗肿瘤功能。因此，一个更好的 了解改变骨骼和免疫动态平衡走向促癌的关键信号机制 这些功能将使针对这种双相效应的新联合疗法的设计成为可能。在这一追求中，我们 已经发现，除了破骨细胞前体的强大激活外，核因子受体激活剂 Kappa-Β配体(RANKL)在M-Ф极化和程序化过程中起着重要的骨免疫纽带作用 死亡配体1(PD-L1)的表达。我们发现BCA细胞中RANKL的升高会引起旁分泌效应 单核细胞向免疫抑制的M2、M、Ф的时空分化初步研究 在本申请中提出的表明，尽管RANKL通过RANK的规范自分泌信号激活了 前馈环，非规范RANKL信号增强M2 MФ和 MDSCs通过富含亮氨酸重复序列的G蛋白偶联受体(LGR4)。 根据我们已公布的和初步的发现，这项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性机制 测试在不干扰肿瘤坏死因子相关的情况下联合应用新的护骨素(OPG)细胞疗法的可能性 凋亡诱导配体(TRAIL)的功能，通过检查点阻断和化疗，逆转肿瘤- 免疫系统和骨骼系统的相关病理学。我们最近采用了一种基于蛋白质结构的 工程方法，并在OPG上确定了用于线索结合的关键区域，并成功开发 并在体内验证了OPG突变体(OPGY49R)，它保留了RANKL结合，但缺乏TRAIL结合。初步 研究，直接比较基于细胞的单应用OPGY49R治疗与多个应用的 中和RANKL单抗治疗表明OPGY49R在单次注射和 与RANKL单抗治疗相比，CD8+T细胞耗竭显著减少。更重要的是，OPGY49R 显著减少体内原发肿瘤的转移，显示出其相对于RANKL的潜在优势 MAb，denosumab，未能延缓高危早期患者的骨转移或疾病复发- 在最近的一项国际双盲随机安慰剂对照的3期研究(D-CARE)中，BCA分期。 这项提议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。 小鼠BCA模型，适用于转移前和转移疾病。