Bioengineered, patient-specific bonemarrow model for studying leukemic niche interactions

项目摘要/摘要 急性白血病是儿童和年轻人中最常见的癌症(约30%)。这个 高危白血病的实验疗法的进展受到了以下方面的限制 永生化细胞系和体内异种移植模型的繁琐性质和有限的吞吐量。在这 背景，缺乏用于原发白血病样本体外培养的强大系统是阻碍 儿童白血病有效遗传和化学筛查的发展。体外系统，包括 工程组织和单芯片器官系统对干细胞和癌症越来越感兴趣 这些领域是人类特有的疾病研究和治疗测试平台。骨骼的体外模型 骨髓(BM)尚未获得发展势头，这主要是由于它们的吞吐量减少、生物方面的技术障碍 研究，以及起始基质细胞群体的异质性。此外，只有很少的尝试 在工程系统中培养原代供者来源的恶性血细胞，使患者特异性 对疾病的研究。在这个拟议的项目中，我将(目标1)设计一种人类诱导多能干细胞(IPSC)- 衍生骨髓组织模型，由成骨细胞、间充质基质细胞和内皮细胞组成 在骨支架内，用于在体外维持急性淋巴细胞白血病(ALL)的表型。然后我将使用 此模型旨在研究恶性ALL细胞的分泌体或ALL细胞自身如何相互作用 在工程模型中，使用健康的造血干/祖细胞(HSPC)和健康的基质。 我推测，一个能够在体外支持HSPC的工程化人类骨髓微环境将 保持所有原始细胞的表型比单层培养或患者更接近未处理的样本 衍生的异种移植模型，进一步研究淋巴母细胞与健康人的直接和间接相互作用 骨髓。众所周知，急性白血病会改变它们的微环境，而且在 许多情况下，在治疗过程中使用间质来保护恶性克隆；我假设这个模型系统 将能够更好地预测治疗反应，识别所有潜在的耐药机制。我们的 Lab带来了在人体组织工程、IPSC技术和治疗测试方面的强大专业知识，以及 在造血、癌症和测序专家的支持下，我相信拟议的项目将成功 建立一种研究恶性白血病转化过程中人骨髓的新工具 对治疗的抗拒。

PROJECT SUMMARY / ABSTRACT Acute leukemias represent the most frequent group of cancer (~30%) in children and young adults. The advancement of experimental therapeutics for high risk leukemias has been limited by the inadequacy of immortalized cell lines and the cumbersome nature and limited throughput of in vivo xenograft models. In this context, the lack of robust systems for in vitro culture of primary leukemia samples is a significant barrier for the development of effective genetic and chemical screens in pediatric leukemia. In vitro systems, including engineered tissues and organ-on-a-chip systems, are gaining increased interest in the stem cell and cancer fields as human-specific platforms for the study of disease and therapeutic testing. In vitro models of the bone marrow (BM) have yet to gain momentum, largely due to their reduced throughput, technical barriers in biological research, and the heterogeneity of starting stromal cell populations. Further, there have been only few attempts to culture primary donor-derived malignant blood cells in engineered systems, which enable patient-specific studies of disease. In this proposed project, I will (Aim 1) engineer a human, induced pluripotent stem cell (iPSC)- derived bone marrow tissue model, comprised of osteoblasts, mesenchymal stromal cells, and endothelial cells within a bone scaffold, for maintenance of acute lymphoblastic leukemia (ALL) phenotype in vitro. I will then use this model to (Aim 2) study how the secretome of malignant ALL blasts, or the ALL blasts themselves, interact with both healthy hematopoietic stem and progenitor cells (HSPCs) and healthy stroma in the engineered model. I hypothesize that an engineered human BM microenvironment, capable of supporting HSPCs in vitro, will maintain the phenotype of ALL blasts closer to unmanipulated samples than monolayer cultures or patient derived xenograft models, further enabling studies of direct and indirect lymphoblast interactions with the healthy bone marrow. It has been well established that acute leukemias alter their microenvironmental niche, and in many cases, use the stroma to protect malignant clones during treatment; I hypothesize that this model system will be better able to predict therapeutic responses, identifying potential mechanisms of resistance in ALL. Our lab brings strong expertise in engineering human tissues, iPSC technologies, and therapeutic testing, and with support of experts in hematopoiesis, cancer, and sequencing, I believe that the proposed project will successfully establish a novel tool for studying the human bone marrow during malignant leukemic transformation and resistance to therapy.