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

基本信息

批准号：
9752259
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
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9752259
关键词：
Address Adult Affect Antigens Autologous Benchmarking Biological Assay Biological Models Biology Biopsy Bone Marrow CD34 gene CD8-Positive T-Lymphocytes CSF3 gene CTLA4 gene Cells Clinical Data Clinical Trials 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 Nivolumab Pathway interactions Patient Education Patients Plasma Play Predisposition Refractory Resistance Role SLEB2 gene Signal Transduction Source System T-Lymphocyte Testing Therapy trial Thymic Tissue Thymus Gland Time Transplantation Treatment Efficacy Tumor Cell Line Validation anti-tumor immune response cancer cell cohort cytokine drug efficacy experimental study hematopoietic differentiation host neoplasm interaction human disease human model humanized mouse improved improved outcome in vivo in vivo Model induced pluripotent stem cell inhibitor/antagonist macrophage melanoma mouse model neoplastic cell novel strategies patient response personalized medicine precursor cell predicting response prevent prospective reconstitution resistance mechanism response reverse genetics self-renewal therapy outcome treatment response tumor tumor growth

项目摘要

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.

概括研究人类癌症的一个限制是具有适当的人类基质和模型的可用性有限免疫。鉴于免疫系统在黑色素瘤中发挥的关键作用，这一警告对于黑色素瘤尤其重要。它的发展。为了解决这些局限性，我们计划以人性化的方式研究黑色素瘤生物学和治疗小鼠（HM）模型。我们最初开发了一种错配的HM（mHM；肿瘤和造血干细胞 [HSCs] 来自不同来源），现在已经生成了自体 HM（aHM；肿瘤、HSCs 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归巢到正在生长的肿瘤中，并迁移到预存在于小鼠基质中，并与人类癌细胞相互作用。肿瘤在 HM 中生长得更紧密与非 HM 小鼠中生长的肿瘤相比，其与起源肿瘤相似，并且由以下因素引起的基因表达漂移之前的传代被部分逆转。关键免疫和基质途径中的信号传导更为突出并且与 HM 模型和非 HM 模型中的原始患者非常相似。免疫细胞有效安装由包括 T 细胞在内的人类免疫细胞介导的肿瘤特异性免疫反应。重要的是，免疫 HM 黑色素瘤模型中免疫治疗的反应显示出与患者的治疗结果。两个突出且尚未充分研究的问题限制了 HM 的更广泛应用：肿瘤与免疫匹配的程度，会影响免疫反应的真实性，进而影响免疫反应的准确性。只有通过比较 mHM 和 aHM 才能理解。对 HM 的第二个批评是人类 T 的异种教育小鼠胸腺细胞上的细胞，这是由于此类模型中缺乏功能性人类胸腺所致。在这个应用程序中，我们将进一步解决自体与不匹配 HM 的相关性，以测试哪个该方法产生了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从患者身上重建功能性胸腺。该提案的总体目标是开展联合临床试验，我们将完成以下任务：1）前瞻性地生成和表征 aHM 和来自 20 名黑色素瘤患者的 mHM，2) 测试哪种方法产生更具代表性的黑色素瘤 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[CTLA4]和程序化细胞）每位患者接受的死亡蛋白 1 [PD-1] 抑制剂）并与临床数据相关联，4) 确定 CTLA4/PD-1抑制剂耐药机制；最后作为选定的探索性目标在这种情况下，我们将 5) 从同一患者的 HSC 中生成胸腺上皮，这将导致 HM 具有完全的自体黑色素瘤、胸腺和免疫系统，从而能够在严格的人类环境中进行免疫细胞教育语境。该项目将增进我们对黑色素瘤和人类肿瘤与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，导致发现改善个性化治疗和改善结果的新方法。