Development of an autologous humanized model of melanoma exploring human thymic education capacity

开发黑色素瘤自体人源化模型，探索人类胸腺教育能力

• 批准号: 9386506
• 负责人: Antonio Jimeno
• 金额: $ 58.08万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9386506
• 关键词: Address; Adult; Affect; Antigens; Apoptosis; Autologous; Benchmarking; Biological Assay; Biological Models; Biology; Biopsy; Bone Marrow; CD34 gene; CD8-Positive T-Lymphocytes; CSF3 gene; Cells; Clinical Data; Clinical Trials; Cytotoxic T-Lymphocyte-Associated Protein 4; Data; Development; Education; Engraftment; Enrollment; Epithelium; Exposure to; Fibrous capsule of kidney; Gene Expression; Genetic; Genetic Drift; Goals; Graft vs Tumor Effect; Growth; Hematopoietic stem cells; Home environment; Human; Immune; Immune Targeting; Immune checkpoint inhibitor; Immune response; Immune system; Immunity; Immunotherapy; Implant; Malignant Neoplasms; Mediating; Melanoma Cell; Mesenchymal; Mesenchymal Stem Cells; Modeling; Monitor; Morphology; Mus; Natural Killer Cells; Pathway interactions; Patient Education; Patients; Pharmaceutical Preparations; Plasma; Play; Predisposition; Proteins; Refractory; Resistance; Role; Signal Transduction; Source; System; T-Lymphocyte; Testing; Therapy trial; Thymic Tissue; Thymus Gland; Time; Transplantation; Treatment Efficacy; Tumor Cell Line; Validation; cancer cell; cohort; cytokine; data modeling; drug efficacy; experimental study; host neoplasm interaction; human disease; humanized mouse; improved; improved outcome; in vivo; in vivo Model; induced pluripotent stem cell; inhibitor/antagonist; macrophage; melanoma; mouse model; neoplastic cell; novel strategies; personalized medicine; precursor cell; predicting response; prevent; prospective; reconstitution; resistance mechanism; response; reverse genetics; self-renewal; therapy outcome; treatment response; tumor; tumor growth

SUMMARY A constraint to studying human cancer is the limited availability of models with appropriate human stroma and immunity. This caveat is particularly relevant for melanoma, given the pivotal role the immune system plays in its development. To address these limitations, we plan to study melanoma biology and therapy in a humanized mouse (HM) model. We initially developed a mis-matched HM (mHM; tumor and hematopoietic stem cells [HSCs] from different sources), and now have generated an autologous HM (aHM; tumor, HSCs and mesenchymal stem cells [MSCs] from the same patient). In HM bearing tumors the human immune and mesenchymal cells from the patient's bone marrow homed into the growing tumor, migrated into the pre- existing mouse stroma, and interacted with the human cancer cells. Tumors grown in HM more closely resembled the originator tumor than those grown in non-HM mice, and the drift in gene expression caused by prior passaging was partially reversed. Signaling in key immune and stroma pathways was more prominent and closely resembled the originator patient in HM vs. non-HM models. The immune cells mounted effective tumor-specific immune responses, mediated by human immune cells including T cells. Significantly, immune responses upon immune therapies in the HM melanoma models showed a correlation with the patient's therapy results. Two salient and under-studied issues limit the wider application of HM: The relevance of the degree of tumor and immune matching, which can affect the faithfulness of immune response, and that can only be appreciated by comparing mHM and aHM. A second criticism to HM is xenogenic education of human T cells on mouse thymic cells, resulting from the lack of a functional human thymus in such models. In this application we will further address the relevance of an autologous versus mis-matched HM to test which approach gives rise to a more representative model of melanoma; secondly, using the same precursor cells from the patient we will reconstitute a functional thymus. The overarching goal of this proposal is to conduct a co-clinical trial where we will accomplish the following: 1) prospectively generate and characterize aHM and mHM from 20 melanoma patients, 2) test which approach yields a more representative melanoma HM model, 3) test in HM the immune drugs (cytotoxic T-lymphocyte-associated protein 4 [CTLA4] and programmed cell death protein 1 [PD-1] inhibitors) that each patient received and correlate with the clinical data, 4) identify the mechanisms involved in resistance to CTLA4/PD-1 inhibitors; and finally as an exploratory Aim in selected cases, we will 5) generate thymic epithelium from the same patient's HSCs that will result in HM with a fully autologous melanoma, thymus and immune system, thus enabling immune cell education in a strictly human context. This project will advance our understanding of the tumor-host interaction in melanoma and human cancer, by better characterizing the interplay between melanoma cells and the immune and stroma systems, leading to the discovery of new approaches to improve personalization of therapy and improve outcomes.

总结 研究人类癌症的一个限制是具有适当人类基质的模型的有限可用性， 免疫力考虑到免疫系统在黑色素瘤中的关键作用，这一警告与黑色素瘤特别相关。 它的发展。为了解决这些局限性，我们计划在人源化的细胞中研究黑色素瘤的生物学和治疗。 小鼠（HM）模型。我们最初开发了一种不匹配的HM（mHM;肿瘤和造血干细胞）， [HSC]来自不同来源），并且现在已经产生了自体HM（aHM;肿瘤，HSC和 来自同一患者的间充质干细胞（MSC））。在携带HM的肿瘤中，人类免疫和 来自患者骨髓的间充质细胞进入生长的肿瘤，迁移到前 存在的小鼠基质，并与人类癌细胞相互作用。在HM中生长的肿瘤 与非HM小鼠中生长的肿瘤相似， 先前的传代被部分逆转。在关键免疫和基质通路中的信号传导更为突出 并且在HM与非HM模型中与原始患者非常相似。免疫细胞有效地 肿瘤特异性免疫应答，由包括T细胞在内的人免疫细胞介导。重要的是，免疫 在HM黑色素瘤模型中，免疫治疗后的反应显示出与患者的 治疗结果。两个突出的和研究不足的问题限制了HM的更广泛应用： 肿瘤和免疫匹配的程度，这可能会影响免疫反应的忠实性， 只能通过比较mHM和aHM来理解。对HM的第二个批评是人类T的异种教育 在小鼠胸腺细胞上的细胞，这是由于在这种模型中缺乏功能性的人胸腺。在这 应用中，我们将进一步解决自体与错配HM的相关性，以测试 这种方法产生了一个更具代表性的黑色素瘤模型;其次，使用相同的前体细胞， 我们将重建一个功能正常的胸腺本提案的总体目标是开展一项 联合临床试验，我们将完成以下任务：1）前瞻性地生成和表征aHM， 来自20名黑素瘤患者的mHM，2）测试哪种方法产生更有代表性的黑素瘤HM模型， 3)在HM中测试免疫药物（细胞毒性T淋巴细胞相关蛋白4 [CTLA 4]和程序性细胞 死亡蛋白1 [PD-1]抑制剂），并与临床数据相关联，4）确定 CTLA 4/PD-1抑制剂耐药相关机制;最后，作为探索性目的， 例，我们将5）从同一患者的HSC中产生胸腺上皮，这将导致HM具有完全的 自体黑色素瘤，胸腺和免疫系统，从而使免疫细胞教育在一个严格的人类 上下文该项目将促进我们对黑色素瘤和人类肿瘤-宿主相互作用的理解。 癌症，通过更好地表征黑色素瘤细胞与免疫和基质系统之间的相互作用， 从而发现新的方法来改善治疗的个性化和改善结果。