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

用于研究白血病生态位相互作用的生物工程、患者特异性骨髓模型

• 批准号: 10536104
• 负责人: Daniel Naveed Tavakol
• 金额: $ 4.68万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-13 至 2023-07-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10536104
• 关键词: Accounting; Acute Lymphocytic Leukemia; Acute leukemia; Adherent Culture; Adult; Apoptosis; Apoptotic; BCL2 gene; Biological Assay; Biological Models; Biomedical Engineering; Blood Cells; Bone Marrow; CXCL12 gene; Cancer Model; Cell Line; Cells; Cessation of life; Chemicals; Child; Childhood Leukemia; Coculture Techniques; Development; Disease; Disease model; Drug resistance; Endothelial Cells; Engineering; Engraftment; Epigenetic Process; Experimental Leukemia; Exposure to; Fibrin; Flow Cytometry; Genetic; Goals; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; Heterogeneity; Homeostasis; Human; Human Engineering; Hydrogels; IL7 gene; In Vitro; Interleukin-1 beta; Interleukin-6; Investigational Therapies; Lymphoblastic Leukemia; Lymphoid; Maintenance; Malignant - descriptor; Malignant Bone Neoplasm; Malignant Neoplasms; Marrow; Mesenchymal; Metabolic; Methodology; Modeling; Myelogenous; Nature; Organoids; Osteoblasts; Outcome; PTPRC gene; Patients; Pharmaceutical Preparations; Phenotype; Physiology; Play; Population; Prediction of Response to Therapy; Predictive Value; Research; Role; Sampling; Solid Neoplasm; Stromal Cells; Study models; Supplementation; System; TNF gene; Testing; Therapeutic; Therapeutic Human Experimentation; Tissue Engineering; Tissue Model; Tissues; Treatment Efficacy; Trichrome stain method; Tropism; Work; Xenograft Model; Xenograft procedure; acute lymphoblastic leukemia cell; biological research; bone engineering; bone scaffold; bone sialoprotein; cancer type; cell immortalization; cell stroma; cell type; cytokine; efficacy testing; high risk; human model; human tissue; improved; in vitro Model; in vivo; induced pluripotent stem cell; induced pluripotent stem cell technology; interest; leukemia; leukemic transformation; lymphoblast; mesenchymal stromal cell; microCT; molecular marker; monolayer; neoplastic cell; novel; novel therapeutics; organ on a chip; patient derived xenograft model; progenitor; resistance mechanism; response; response to injury; stem; stem cells; therapeutic development; therapeutic evaluation; therapy resistant; tool; transcriptomics; tumor; young adult

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.

项目概要/摘要 急性白血病是儿童和年轻人中最常见的癌症（约 30%）。这 高危白血病实验疗法的进展因缺乏足够的技术而受到限制 永生化细胞系以及体内异种移植模型的繁琐性质和有限的通量。在这个 在此背景下，缺乏强大的原发性白血病样本体外培养系统是该领域的一个重大障碍。 开发针对儿童白血病的有效遗传和化学筛查。体外系统，包括 工程组织和器官芯片系统越来越受到人们对干细胞和癌症的兴趣 领域作为疾病研究和治疗测试的人类特定平台。体外骨模型 骨髓 (BM) 尚未获得发展动力，这主要是由于其吞吐量降低、生物技术壁垒 研究以及起始基质细胞群的异质性。此外，只有很少的尝试 在工程系统中培养原代供体来源的恶性血细胞，从而实现患者特异性 疾病研究。在这个拟议的项目中，我将（目标 1）设计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）表型。然后我将使用 该模型（目标 2）研究恶性 ALL 原始细胞的分泌组或 ALL 原始细胞本身如何相互作用 工程模型中含有健康的造血干细胞和祖细胞 (HSPC) 以及健康的基质。 我假设，能够在体外支持 HSPC 的工程化人类 BM 微环境将 保持 ALL 原始细胞的表型比单层培养物或患者更接近未操作的样品 衍生的异种移植模型，进一步支持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，急性白血病会改变其微环境生态位，并且 许多情况下，在治疗期间使用基质来保护恶性克隆；我假设这个模型系统 将能够更好地预测治疗反应，识别 ALL 的潜在耐药机制。我们的 实验室在人体组织工程、iPSC 技术和治疗测试方面拥有丰富的专业知识，并且 在造血、癌症和测序专家的支持下，我相信该项目将成功 建立一种研究恶性白血病转化过程中人类骨髓的新工具 对治疗的抵抗。