Evolutionary dynamics of melanoma metastasis

黑色素瘤转移的进化动力学

• 批准号: 8568851
• 负责人: Richard Mark White
• 金额: $ 265.87万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8568851
• 关键词: Address; Adult; Alleles; Biological Assay; Cancer Model; Cessation of life; DNA biosynthesis; DNA-Directed DNA Polymerase; Disease; Elements; Evolution; Family; Fishes; Gene Transfer Techniques; Genetic Screening; Genomics; Genotype; Goals; Laboratories; Lead; Lesion; Malignant Neoplasms; Maps; Mediating; Metastatic Melanoma; Methods; Modeling; Mutation; Natural Selections; Neoplasm Metastasis; Organism; Patients; Primary Neoplasm; Solutions; Stress; System; Testing; Time; Work; Zebrafish; base; cancer cell; cellular imaging; exome sequencing; loss of function; melanocyte; melanoma; neoplastic cell; novel; pressure; progenitor; stressor; tumor

DESCRIPTION (provided by applicant): Metastatic disease remains the defining feature of advanced malignancy, and is responsible for the vast majority of cancer deaths. Metastasis can be conceptualized as an evolutionary landscape, composed of key elements of Darwinian evolution: heritable (epi)genotypes, geographic dispersal, and novel microenvironmental selection pressures. I hypothesize that altering these evolutionary landscapes would provide a strikingly new method for treating cancer, in which the cancer cells can co-exist with the host over long periods of time. To achieve this requires a deep mechanistic understanding of the ways in which tumors generate novel genotypes, and how natural selection in the microenvironment amplifies these mutations. My work utilizes the zebrafish, a small vertebrate organism that has only recently come to light as an important cancer model. The zebrafish offers several unique capacities for studying metastasis: high-throughput transgenesis, unbiased genetic screens, and single cell imaging in the optically transparent casper adult fish. For the period of this proposal, I plan to address three primary questions in metastatic melanoma: 1) can we identify the incipient genetic changes that allow for metastatic progression, whether they arise in the primary tumor or after dissemination, 2) is adaptive mutation required for metastasis, and can this be modified?, and 3) can we identify host microenvironments that disfavor metastatic progression? To do this, I will build upon a zebrafish model of melanoma in which the BRAFV600E allele is expressed in melanocyte progenitors, in the context of p53 loss of function (the BRAFV600E;p53-/- model). By combining the "brainbow" fate mapping system with the BRAFV600E;p53-/- fish, I will use exome sequencing to identify genomic lesions associated with metastasis based on lineage, space and time. These candidate changes can be functionalized using a metastasis assay I have developed in the transparent casper strain. I hypothesize that selection stressors during metastasis lead to a state of adaptive mutation, in which the error rate of DNA replication is temporarily increased to find an evolutionary solution to that stress. To test this, I will generate zebrafish with mutation reportes, and then use this system to probe whether adaptive mutation mediated by error-prone Y family DNA polymerases promotes metastatic progression. Finally, since the host microenvironment provides the ultimate selection pressure, I will perform an unbiased genetic screen to identify novel stromal regulators of metastatic progression. Together, these studies provide a comprehensive framework that considers both tumor cell-intrinsic and microenvironmental dynamics of metastatic disease. The long-term goal of my laboratory is to utilize this information to identify therapies which can convert disseminated disease into a stable state, unable to further progress, leading to large improvements in long-term survival in patients with established metastases.

描述（由申请人提供）：转移性疾病仍然是晚期恶性肿瘤的定义特征，并导致绝大多数癌症死亡。转移可以被概念化为一种进化景观，由达尔文进化论的关键要素组成：可遗传的（EPI）基因型，地理散布和新型的微环境选择压力。我假设改变这些进化景观将为治疗癌症提供惊人的新方法，在这种方法中，癌细胞可以长期与宿主共存。为了实现这一目标，需要深入的机理理解肿瘤产生新型基因型的方式，以及微环境中的自然选择如何放大这些突变。我的工作利用了斑马鱼，斑马鱼是一种小型脊椎动物，直到最近才揭示为重要的癌症模型。斑马鱼提供了几种用于研究转移的独特能力：高通量转基因，无偏的遗传筛选和光学透明的卡斯珀成年鱼类中的单细胞成像。在这项提议期间，我计划解决转移性黑色素瘤中的三个主要问题：1）我们能否确定允许转移进展的初步遗传变化，无论它们是在原发性肿瘤中还是在传播后，2）是转移所需的适应性突变，并且可以改变宿主的微心体进展吗？为此，我将建立在黑色素瘤模型的基础上，其中BRAFV600E等位基因在黑素细胞祖细胞中表达，在p53功能丧失的背景下（BRAFV600E； p53 - / - 模型）。通过将“ Brainbow”命运映射系统与BRAFV600E; p53 - / - FISH相结合，我将使用外显子组测序来识别基于谱系，空间和时间的转移相关的基因组病变。这些候选变化可以使用我在透明的Casper菌株中开发的转移测定法进行功能化。我假设转移期间的选择应力源导致适应性状态 突变，其中DNA复制的错误率暂时增加，以找到应力的进化解决方案。为了测试这一点，我将通过突变报告生成斑马鱼，然后使用该系统来探测由易受错误的Y家族DNA聚合酶介导的适应性突变是否促进转移性进展。最后，由于宿主微环境提供了最终的选择压力，因此我将执行一个无偏的遗传筛选，以识别转移性进展的新型基质调节剂。总之，这些研究提供了一个全面的框架，该框架考虑了转移性疾病的肿瘤细胞内部和微环境动力学。我实验室的长期目标是利用这些信息来识别可以将传播疾病转化为稳定状态的疗法，无法进一步进步，从而导致既定转移患者的长期生存率大大提高。

项目成果

Rational Design of Polyglutamic Acid Delivering an Optimized Combination of Drugs Targeting Mutated BRAF and MEK in Melanoma.

合理设计聚谷氨酸，提供针对黑色素瘤中突变 BRAF 和 MEK 的优化药物组合。

• DOI: 10.1002/adtp.202000028
• 发表时间: 2020
• 期刊: Advanced therapeutics
• 影响因子: 4.6
• 作者: Pisarevsky,Evgeni;Blau,Rachel;Epshtein,Yana;Ben-Shushan,Dikla;Eldar-Boock,Anat;Tiram,Galia;Koshrovski-Michael,Shani;Scomparin,Anna;Pozzi,Sabina;Krivitsky,Adva;Shenbach-Koltin,Gal;Yeini,Eilam;Fridrich,Lidar;White,Richard;Satchi-
• 通讯作者: Satchi-