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A novel in vivo model system of melanoma using a human-mouse neural crest cell chimera

使用人-小鼠神经嵴细胞嵌合体的新型黑色素瘤体内模型系统

基本信息

批准号：
9230815
负责人：
Katherine Jean Wert
金额：
$ 1.7万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9230815
关键词：
Adolescent Age Age-Years American Area Biological Models Cancer Biology Cancer Etiology Cell Survival Cells Cessation of life Chimera organism Complex Coupled Cues Development Differentiation Antigens Disease Disease model Drug Modelings Drug toxicity Embryo Embryonic Development Environment Fertility Generations Genes Goals Human Human Development Immune In Vitro Individual Injectable Injection of therapeutic agent Knowledge Label Laboratories Location Malignant Neoplasms Melanoma Cell Metastatic Melanoma Methods Microinjections Microscopy Modeling Multipotent Stem Cells Mus Neoplasm Metastasis Neural Crest Neural Crest Cell Organism Outcome Phenotype Pigmentation physiologic function Play Population Pre-Clinical Model Preclinical Drug Evaluation Predisposition Pregnancy Proto-Oncogene Protein c-kit Risk Role Shapes Signal Transduction Stem cells Study models Subcutaneous Injections System Technology Testing Therapeutic Agents Time Tissues Transfection Transgenic Organisms Transplantation Tumor Cell Invasion Tumor-Derived cell motility disease phenotype experimental analysis human disease implantation in utero in vivo in vivo Model induced pluripotent stem cell innovation melanocyte melanoma migration mouse model neoplastic cell neural plate novel novel strategies overexpression patient population public health relevance stem therapeutic evaluation therapeutic target time use tool tumor vertebrate embryos young adult

项目摘要

DESCRIPTION (provided by applicant): Metastatic melanoma is one of the fastest growing cancers, causing more than 8,650 deaths in 2009 alone, and this number is projected to increase over time. Models for the study of melanoma have multiple limitations, including the inability of most transgenic systems to accurately reproduce complex disease phenotypes such as cancer, and the inability of human embryonic stem (hES) and human induced pluripotent stem (hiPS) cell technology to reproduce the host microenvironments. It has been shown, however, that aggressive melanoma cells up-regulate a subset of neural crest (NC) cell guidance and differentiation markers as well as being a NC cell-derived cancer. We hypothesize that an in vivo platform for the study of melanoma development, invasion, and metastasis can be created using human NC cell chimeras. There are two specific goals of this study: 1) to optimize the incorporation of hES and hiPS cell-derived NC cells into a murine system, and 2) to model human melanoma development and metastasis in vivo. These chimeras will be created by in utero injections of hES or hiPS cell-derived NC cells into the developing mouse embryo at embryonic day 8.5 (E8.5). Human NC donor cells will be labeled with the eGFP marker and injected embryos will be analyzed during gestation and post-natally to localize the injected cells and establish methods optimizing their survival and colonization, such as the over-expression of NC cell survival genes via lentiviral transfection. We will then utilize this chimeric system to over-express common genes with known roles in melanoma, and inject these NC cells in utero at E8.5 to model melanoma initiation and development in vivo in a murine model system. Additionally, we will promote tumor formation of the hES or hiPS cell-derived NC cells after subcutaneous injection into an immune-compromised mouse, and the tumor cells formed can be cultured and re-injected at E8.5 into the developing mouse embryo to study tumor migration and metastasis over the developmental period. Overall, we propose to bridge the studies of embryonic development with cancer development, focusing on melanoma. We will analyze the progression of melanoma using hES/hiPS cell technology coupled with in vivo murine modeling. This novel system will not only bridge the knowledge of embryonic development and cancer biology, but will provide an in vivo model of melanoma to be used for testing potential therapeutic agents.

描述（由申请人提供）：转移性黑色素瘤是增长最快的癌症之一，仅 2009 年就导致超过 8,650 人死亡，并且预计这一数字会随着时间的推移而增加。研究黑色素瘤的模型具有多种局限性，包括大多数转基因系统无法准确再现癌症等复杂疾病表型，以及人类胚胎干（hES）和人类诱导多能干（hiPS）细胞技术无法再现宿主微环境。然而，研究表明，侵袭性黑色素瘤细胞上调神经嵴 (NC) 细胞引导和分化标记物的子集，并且是一种 NC 细胞衍生的癌症。我们假设可以使用人类 NC 细胞嵌合体创建用于研究黑色素瘤发育、侵袭和转移的体内平台。这项研究有两个具体目标：1) 优化 hES 和 hiPS 细胞衍生的 NC 细胞与小鼠系统的整合，2) 模拟人类黑色素瘤的体内发育和转移。这些嵌合体将通过在胚胎第 8.5 天（E8.5）在子宫内将 hES 或 hiPS 细胞衍生的 NC 细胞注射到发育中的小鼠胚胎中而产生。人类 NC 供体细胞将用 eGFP 标记进行标记，注射的胚胎将在妊娠期间和出生后进行分析，以定位注射的细胞并建立优化其生存和定植的方法，例如通过慢病毒转染过度表达 NC 细胞生存基因。然后，我们将利用该嵌合系统过度表达在黑色素瘤中具有已知作用的常见基因，并在 E8.5 将这些 NC 细胞注入子宫内，以在小鼠模型系统中模拟体内黑色素瘤的起始和发展。此外，我们将hES或hiPS细胞衍生的NC细胞皮下注射到免疫受损的小鼠体内后促进肿瘤形成，形成的肿瘤细胞可以在E8.5培养并重新注射到发育中的小鼠胚胎中，以研究发育期间的肿瘤迁移和转移。总体而言，我们建议将胚胎发育研究与癌症发育研究联系起来，重点关注黑色素瘤。我们将使用 hES/hiPS 细胞技术结合体内小鼠模型来分析黑色素瘤的进展。这种新颖的系统不仅将在胚胎发育和癌症生物学知识之间架起桥梁，而且将提供黑色素瘤的体内模型，用于测试潜在的治疗药物。