Biomarkers and Therapeutic Targets in Tumor Microenvironment and Metastasis

肿瘤微环境和转移中的生物标志物和治疗靶点

• 批准号: 10486817
• 负责人: Rosandra Kaplan
• 金额: $ 95.43万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10486817
• 关键词: Adjuvant; Adjuvant Therapy; Adult; Aging; Area; Biological; Biological Assay; Biological Markers; Biological Models; Blood; Blood Vessels; Bone Marrow; Bone Marrow Neoplasms; CD8-Positive T-Lymphocytes; CSF1R gene; Cell Communication; Cell Line; Cell Lineage; Cell Therapy; Cells; Chemotherapy and/or radiation; Childhood; Chronic Disease; Clinical; Collaborations; Colony-Forming Units Assay; Colony-Stimulating Factors; Cytotoxic T-Lymphocytes; Data; Development; Disease; Disseminated Malignant Neoplasm; Distant; Effectiveness; Endothelial Cells; Endothelium; Engineering; Enrollment; Epigenetic Process; Ewings sarcoma; Excision; Extracellular Matrix; FLT3 gene; Fibrinogen; Fibroblasts; Flow Cytometry; Gene Expression Profile; Genetic; Genetic Engineering; Genetic Transcription; Giant Cell Tumors; Growth; Growth and Development function; Heart Diseases; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; High-Risk Cancer; Human; Image; Immune; Immune Targeting; Immune response; Immune system; Immunofluorescence Immunologic; Immunosuppression; Immunotherapy; Individual; Inflammatory; Institutional Review Boards; Investigation; Laboratories; Lead; Li-Fraumeni Syndrome; Longevity; Longitudinal Studies; Luciferases; Macrophage Colony-Stimulating Factor; Malignant Neoplasms; Measures; Mediating; Mediator of activation protein; Mesenchymal; Modeling; Molecular; Monitor; Mus; Mutation; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; NF1 gene; Neoplasm Metastasis; Neurofibrosarcoma; Normal tissue morphology; Organ Transplantation; Outcome; Patients; Pericytes; Phase; Play; Plexiform Neurofibroma; Population; Pre-Clinical Model; Primary Neoplasm; Process; RNA; Recurrence; Relapse; Rhabdomyosarcoma; Risk; Role; Sampling; Signal Transduction; Site; Stromal Cells; Supporting Cell; Syndrome; System; TP53 gene; Testing; Therapeutic; Tissues; Translating; Translational Research; Transplant-Related Disorder; Treatment Efficacy; Tumor Cell Line; Tumor Immunity; Tumor Markers; Vascular Diseases; Work; base; bone circulation; cancer predisposition; cell behavior; cell type; childhood sarcoma; clinical investigation; conventional therapy; cytotoxicity; endothelial stem cell; experimental study; fighting; hematopoietic stem cell niche; high risk; improved; in vitro Model; in vivo; individualized medicine; inhibitor/antagonist; insight; macrophage; mast cell; microvesicles; monocyte; myeloid cell development; neoplastic cell; non-genetic; novel; osteosarcoma; pediatric patients; pre-clinical; preclinical study; predictive tools; progenitor; prognostic tool; programs; rare cancer; receptor; recruit; repository; response; single cell sequencing; specific biomarkers; standard care; stem cells; targeted treatment; therapeutic target; therapy design; therapy development; tissue regeneration; tumor; tumor microenvironment; tumor progression; tumor xenograft; tumorigenesis; wound healing

As one of the crucial steps in metastatic progression requires tumor to successfully interact with its local microenvironment, it follows that targeting this cross-talk may be an attractive adjuvant to standard treatment approaches. We are currently focused on developing therapies that target the associated tumor recruited host immmune and stromal cells. We have an IRB approved biological repository study to obtain blood, bone marrow, tumor and adjacent normal tissue when available from patients with malignancy and healthy donors. We continue our on-going studies of measuring and characterizing the circulating bone marrow-derived progenitor, immune, endothelial and mesenchymal cells that may be altered in the setting of cancer and other chronic diseases. Utilizing both quantification and functional assays, including flow cytometry and colony forming unit assays, we are assessing the circulating bone marrow-derived progenitor cell populations in pediatric and adult patients with malignancies. We have broadened our investigations to better understand the changes in the hematopoietic stem cell niche that results in alterations in immune milieu in response to a growing primary tumor. These studies now include in addition to monitoring hematopoietic and endothelial progenitor cells but also CD4 and CD8 T cells and myeloid cells including MDSCs and M1 and M2 macrophages and stromal cell populations. Furthermore, we measure circulating microvesicles released by tumor cells and associated tumor hematopoietic and stromal cells that may impact important cell behavior and are known to be critical to cell-cell communication. We have on-going investigations as to which cells make which microvesicles and their particular content and determining which would be most useful as a biomarker for metastatic risk. Our recent studies have determined host cell plasticity and cell state determine the microvesicles released from these cells and this plasticity in perivascular cells play key roles in regulating metastasis. We are currently investigating markers of this perivascular cell plasticity as a predictor of metastasis and response to conventional therapies and immune based therapies. We continue our collaboration with Dr. Sharon Savage to examine circulating bone marrow-derived cell populations in patients with Li Fraumeni syndrome, which is a high-risk cancer predisposition syndrome related to loss of tumor suppressor p53. We have also developed assays to examine biological functional of specific cells and correlates that can be measured in stored RNA samples in order to correlate outcome data with these biomarkers for metastatic risk. We have established a pre-clinical model system for testing microenvironment-targeting therapy in pediatric sarcomas. Utilizing a Ewings sarcoma (EWS) xenograft tumor cell line and two syngeneic models- rhabdomyosarcoma (RMS) cell line and an osteosarcoma (OS) cell line we have performed flow cytometry and immunofluorescence to demonstrate the influx of myeloid cells and alterations in stromal cell populations in the tumor and pre-metastatic tissues. We also monitor metastatic progression in a resection model using luciferase imaging. In this fashion, pre-metastatic, metastatic colonization and progression to visible metastasis can be followed and compared in treated and untreated groups without requiring multiple terminal end points. We are conducting pre-clinical investigations utilizing inhibitors targeting stromal cell plasticity specifically to assess impact on metastatic progression. We also now have a marker of tumor associated fibroblast activation and stromal cell lineage tracing mice in order to monitor activation of these cells in this process. We have performed serial in vivo mouse experiments examining targeting of myeloid cells and stromal cells to determine their impact on metastatic progression. We are using different investigational agents to determine their specific impact on each microenvironmental cell. These pre-clinical studies will answer whether this approach to treatment may likely be a good window for targeting the recruitment of these microenvironment tumor-associated cells that support tumor progression. Our studies using a colony stimulating factor -one receptor (csf1-R) inhibitor revealed that these models of RMS and Ewings sarcoma secrete a good deal of CSF-1 and lead to the recruitment of CSF-1R expressing cells. These cells are found in early metastatic sites and are immune suppressive and can protect disseminated tumor cells from cytotoxic T cell activity. We are working on developing approaches to promoting myeloid based anti-tumor immunity and performing studies to determine the critical effectors of this cytotoxicity. We have also established a good in vitro model to understand the role of tumor-secreted factors on myeloid cell development and stromal cell plasticity and function and investigating different approaches to targeting this process. These studies allow for investigating function of potential therapeutic inhibitors of the myeloid skewing and polarization process and the activation and expansion of specific perivascular cell populations that promote metastasis. This year we have completed the Phase I portion of Pexidartinib which inihibits FLT3, Kit and CSF1R. We are now planning enrollment on the expanded Phase I in pediatric patients with tenosynovial giant cell tumor, high risk Osteosarcoma, multiply recurrent malignancies in combination with other immunotherapy and Phase II focused on patients with NF1 and plexiform neurofibromas (PN) given a mast cell or other Kit mediated cell- cell crosstalk is thought to promote PN growth and possible progression to malignant peripheral nerve sheath tumors. We are also actively planning two new trials to target immune suppressive myeloid cells. We have developed a new cell therapy based on genetically engineering myeloid cells (GEMys) that can be novel delivery vehicles given their propensity to accumulate in tumor and metastatic sites. These cells can be engineered to deliver Il12 into the tumor milieu and reprogram multiple cell types, change gene transcriptional signatures and reverse immune suppression and enhance anti-tumor immunity. We have developed humanized murine systems to examine human cell therapy with advanced human tumors. On going work to bring this to clinical setting is on-going and harnessing myeloid cells for introduction of signaling in microenvironments. These investigations also include stromal cells and extracellular matrix remodeling. Single cell sequencing can provide exquisite detail of individual cell cluster transcriptional programs. Our laboratory has begun performing single cell sequencing of rare tumors to investigate tumor and microenvironmental interactions. Such studies can be invaluable for tumor and microenvironment genetic and non genetic interactions that provide insights to targeting cross talk and unique aspects of both tumor and associated microenvironment.

由于转移进展的关键步骤之一需要肿瘤与其局部微环境成功相互作用，因此针对这种相互作用可能是标准治疗方法的有吸引力的辅助剂。我们目前专注于开发针对相关肿瘤招募的宿主免疫细胞和基质细胞的疗法。我们进行了 IRB 批准的生物储存库研究，以获取来自恶性肿瘤患者和健康捐赠者的血液、骨髓、肿瘤和邻近正常组织。我们继续进行持续的研究，测量和表征循环骨髓源性祖细胞、免疫细胞、内皮细胞和间充质细胞，这些细胞可能在癌症和其他慢性疾病的情况下发生改变。利用定量和功能测定（包括流式细胞术和集落形成单位测定），我们正在评估患有恶性肿瘤的儿科和成人患者的循环骨髓来源的祖细胞群。我们扩大了研究范围，以更好地了解造血干细胞生态位的变化，这些变化导致免疫环境发生变化，以应对不断生长的原发肿瘤。这些研究现在除了监测造血和内皮祖细胞外，还包括 CD4 和 CD8 T 细胞以及骨髓细胞，包括 MDSC 以及 M1 和 M2 巨噬细胞和基质细胞群。此外，我们还测量肿瘤细胞以及相关肿瘤造血细胞和基质细胞释放的循环微泡，这些微泡可能影响重要的细胞行为，并且已知对细胞间通讯至关重要。我们正在持续研究哪些细胞产生哪些微泡及其特定内容，并确定哪些最适合作为转移风险的生物标志物。我们最近的研究确定了宿主细胞的可塑性和细胞状态决定了这些细胞释放的微泡，血管周围细胞的这种可塑性在调节转移中发挥着关键作用。我们目前正在研究这种血管周围细胞可塑性的标志物，作为转移和对传统疗法和免疫疗法的反应的预测因子。我们继续与 Sharon Savage 博士合作，检查 Li Fraumeni 综合征患者的循环骨髓来源细胞群，这是一种与肿瘤抑制因子 p53 缺失相关的高风险癌症易感综合征。我们还开发了检测特定细胞生物功能的检测方法，并可在存储的 RNA 样本中测量相关性，以便将结果数据与这些转移风险生物标志物相关联。我们建立了一个临床前模型系统，用于测试儿科肉瘤的微环境靶向治疗。利用尤文氏肉瘤 (EWS) 异种移植肿瘤细胞系和两种同基因模型 - 横纹肌肉瘤 (RMS) 细胞系和骨肉瘤 (OS) 细胞系，我们进行了流式细胞术和免疫荧光分析，以证明骨髓细胞的流入和基质细胞群的变化在肿瘤和转移前组织中。我们还使用荧光素酶成像监测切除模型中的转移进展。以这种方式，可以在治疗组和未治疗组中跟踪和比较转移前、转移性定植以及进展为可见转移，而不需要多个终点。我们正在利用针对基质细胞可塑性的抑制剂进行临床前研究，专门评估对转移进展的影响。我们现在还拥有肿瘤相关成纤维细胞激活和基质细胞谱系追踪小鼠的标记，以便监测这些细胞在此过程中的激活。我们进行了一系列体内小鼠实验，检查骨髓细胞和基质细胞的靶向，以确定它们对转移进展的影响。我们正在使用不同的研究药物来确定它们对每个微环境细胞的具体影响。这些临床前研究将回答这种治疗方法是否可能是针对招募这些支持肿瘤进展的微环境肿瘤相关细胞的良好窗口。我们使用集落刺激因子一受体 (csf1-R) 抑制剂进行的研究表明，这些 RMS 和尤文氏肉瘤模型分泌大量 CSF-1，并导致 CSF-1R 表达细胞的募集。这些细胞存在于早期转移部位，具有免疫抑制作用，可以保护播散的肿瘤细胞免受细胞毒性 T 细胞活性的影响。我们正在开发促进基于骨髓的抗肿瘤免疫的方法，并进行研究以确定这种细胞毒性的关键效应子。我们还建立了一个良好的体外模型，以了解肿瘤分泌因子对骨髓细胞发育和基质细胞可塑性和功能的作用，并研究针对该过程的不同方法。这些研究允许研究骨髓偏斜和极化过程的潜在治疗抑制剂的功能以及促进转移的特定血管周围细胞群的激活和扩张。今年我们已经完成了 Pexidartinib 的 I 期部分，该部分抑制 FLT3、Kit 和 CSF1R。我们现在计划扩大 I 期的入组，治疗患有腱鞘巨细胞瘤、高风险骨肉瘤、多发性复发性恶性肿瘤的儿科患者，并与其他免疫疗法相结合，而 II 期则重点关注给予肥大细胞或给予肥大细胞或治疗的 NF1 和丛状神经纤维瘤 (PN) 患者。其他 Kit 介导的细胞间串扰被认为可促进 PN 生长并可能进展为恶性周围神经鞘瘤。我们还积极计划两项针对免疫抑制性骨髓细胞的新试验。我们开发了一种基于基因工程骨髓细胞 (GEMys) 的新细胞疗法，鉴于其在肿瘤和转移部位积聚的倾向，它可以成为新型输送载体。这些细胞可以被设计为将 Il12 传递到肿瘤环境中并重新编程多种细胞类型、改变基因转录特征、逆转免疫抑制并增强抗肿瘤免疫力。我们开发了人源化小鼠系统来检查晚期人类肿瘤的人类细胞疗法。将其引入临床的工作正在进行中，并利用骨髓细胞在微环境中引入信号传导。这些研究还包括基质细胞和细胞外基质重塑。单细胞测序可以提供单个细胞簇转录程序的精致细节。我们的实验室已开始对罕见肿瘤进行单细胞测序，以研究肿瘤和微环境的相互作用。此类研究对于肿瘤和微环境的遗传和非遗传相互作用具有无价的价值，它们为靶向肿瘤和相关微环境的串扰和独特方面提供了见解。