Molecular and functional characterization of the normal and malignant plasma cell – bone interface – from single cell mutual interaction analysis to clinical implications in myeloma bone disease and survival

正常和恶性浆细胞的分子和功能特征 â 骨界面 â 从单细胞相互作用分析到骨髓瘤骨病和生存的临床意义

• 批准号: 401358321
• 负责人: Professorin Dr. Regina Ebert, since 9/2020
• 金额: --
• 依托单位: Hematology and Immunology Research Group
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/401358321?language=en
• 关键词: Molecular; functional; characterization; normal; malignant

Multiple Myeloma is a disease of malignant plasma cells spreading and accumulating in the bone marrow with subsequent induction of bone disease and angiogenesis. These features are in common with other malignant diseases. Multiple myeloma is different in two regards: Myeloma cells (MMC) spread to and live in the same “natural environment” as their physiological counterpart, bone marrow plasma cells (BMPC). Further, as shown by us, MMC rely at least partly on normal plasma cell functions for malignant behaviour when inducing angiogenesis and interacting with bone turnover. Malignant like normal plasma cells depend on receiving survival-support by interaction with other cell types in the bone marrow, collectively called “niche”. In vitro, we have shown in myeloma/ mesenchymal stromal cell (MSC)//osteoblast co-cultures that this interaction is prominently exerted by physical interaction and leads to changes in expression and behaviour in both cells types. We hypothesize that this physical interaction creates niches and elicits individual signatures that strengthen adhesion, support tumor cells, and disrupt osteogenesis, clinically leading to bone loss and pathological fractures. Niches may confer dormancy or resistance (minimal residual disease) features to myeloma. We have shown this molecular crosstalk being drugable exemplified by blocking the osteocyte-specific wnt antagonist sclerostin, observing increased osteoblast numbers and bone formation in myeloma bearing mice. We also showed that the contact-induced KISS1R system is a putative target as a theranostic PET tracer.In this application, we will focus on surface molecules featuring the interaction between normal and malignant plasma cells and MSC/osteoblasts. We will use expression analysis including single cell RNA-seq to address population heterogeneity, and functional assessment of interaction by atomic-force microscopy and CRISPR/CAS9 screen with subsequent collaborative in vivo validation in myeloma mouse models. In a second step, our in vitro findings will be related to a large cohort of molecularly characterized malignant plasma cell samples alongside whole-body imaging assessing bone disease, plasma cell infiltration and angiogenesis. We have shown that relation of data at nano-/micro- to large scale clinical data sets can act as a filter for relevance and will allow identification of pre-clinically applicable findings. The first is evidenced by sclerostin being expressed in osteocytes only, in contrast to malignant plasma cells from 630 MM patients; the second by our co-development of the T-cell bispecific antibody EM801/901 against the BMPC/MMC cell-surface molecule BCMA entering clinical trials in Q1/18, building the basis for a separate application for development of a theranostic BCMA-PET-Tracer. We will contribute to a collaborative network for myeloma dissemination, bone disease and angiogenesis related research questions within the µBone-consortium.

多发性骨髓瘤是一种恶性浆细胞在骨髓中扩散和积聚，随后诱发骨病和血管生成的疾病。这些特征与其他恶性疾病相同。多发性骨髓瘤在两个方面是不同的：骨髓瘤细胞（MMC）与它们的生理对口物骨髓浆细胞（BMPC）扩散并生活在相同的“自然环境”中。此外，如我们所示，MMC在诱导血管生成和与骨转换相互作用时，至少部分依赖于正常浆细胞功能的恶性行为。恶性肿瘤与正常浆细胞一样，依靠与骨髓中的其他细胞类型相互作用来获得生存支持，这些细胞类型统称为“生态位”。在体外，我们已经在骨髓瘤/间充质基质细胞(MSC)/成骨细胞共培养中表明，这种相互作用主要通过物理相互作用发挥，并导致两种细胞类型的表达和行为改变。我们假设这种物理相互作用创造了壁龛并引发了个体特征，这些特征加强了粘附，支持肿瘤细胞，破坏了成骨，在临床上导致骨质流失和病理性骨折。小生境可能赋予骨髓瘤休眠或抵抗（最小残留病）特征。通过阻断骨细胞特异性wnt拮抗剂硬化蛋白，我们已经证明这种分子串扰是可药物化的，观察到骨髓瘤小鼠成骨细胞数量和骨形成的增加。我们还发现，接触诱导的KISS1R系统可能是一种治疗性PET示踪剂。在本应用中，我们将重点研究正常和恶性浆细胞与MSC/成骨细胞相互作用的表面分子。我们将使用表达分析，包括单细胞RNA-seq来解决群体异质性，并通过原子力显微镜和CRISPR/CAS9筛选对相互作用进行功能评估，随后在骨髓瘤小鼠模型中进行协作体内验证。在第二步中，我们的体外研究结果将与大量分子表征的恶性浆细胞样本以及评估骨病、浆细胞浸润和血管生成的全身成像相关。我们已经表明，纳米/微观数据与大规模临床数据集的关系可以作为相关性的过滤器，并允许识别临床前应用的发现。第一个证据是，与630名MM患者的恶性浆细胞相比，硬化蛋白仅在骨细胞中表达；第二个是我们共同开发的针对BMPC/MMC细胞表面分子BCMA的t细胞双特异性抗体EM801/901于2018年第一季度进入临床试验，为开发治疗性BCMA- pet示踪剂的单独应用奠定基础。我们将在µbone -consortium中为骨髓瘤传播、骨病和血管生成相关研究问题的合作网络做出贡献。