Somatic mutation(s) and cellular changes in CCM pathogenesis

CCM 发病机制中的体细胞突变和细胞变化

• 批准号: 10220145
• 负责人: Douglas A. Marchuk
• 金额: $ 33.13万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10220145
• 关键词: 1-Phosphatidylinositol 3-Kinase; AKT Signaling Pathway; Blood Vessels; Brain; Cell Compartmentation; Cell Lineage; Cells; Clinical Trials; Clonal Expansion; Code; DNA Resequencing; DNA sequencing; Data; Development; Disease; Doctor of Philosophy; Endothelial Cells; Event; Exons; Funding; Gene Expression Profile; Genes; Genetic Engineering; Genomic DNA; Genotype; Goals; Growth; Hemangioma; Human; Individual; Knowledge; Laboratories; Lesion; Malignant Neoplasms; Modeling; Molecular; Mus; Mutate; Mutation; Normal Cell; Oncogenes; Operative Surgical Procedures; PIK3CA gene; Pathogenesis; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Poison; Population; Proto-Oncogene Proteins c-akt; Publishing; Research Personnel; Resected; Role; Route; Seizures; Signal Pathway; Signal Transduction; Somatic Mutation; Source; Stroke; Testing; Therapeutic; Tissue Banks; Work; base; cell behavior; cerebral cavernous malformations; clinically relevant; drug candidate; drug repurposing; effective therapy; exome sequencing; experience; gene discovery; human tissue; improved; malformation; mouse model; mutant; new therapeutic target; novel; novel therapeutics; programs; recruit; single-cell RNA sequencing; transcriptomics

ABSTRACT - PROJECT 1 Based on discoveries made in the previous cycle of our program project, Project 1 will probe deeply into the cellular and molecular events of CCM pathogenesis. Using lineage tracing of murine CCM lesions we showed that the initial event of lesion genesis is somatic loss of the remaining wild-type copy of the Ccm gene, followed by a clonal expansion of endothelial cells harboring the same somatic mutation, and what appears to be recruitment of additional ECs into the growing lesion – cells that do not harbor the somatic mutation. Secondly, based on discoveries in the Kahn lab for a role for activated PI3 kinase in lesion development in mouse CCM models, we identified activating PIK3CA mutations in more than 50% of human CCM lesions. These collaborative discoveries suggest we have not yet fully understood the molecular and cellular events that contribute to CCM lesion genesis, growth and maturation. In the first Aim, we will deeply sequence bulk human CCMs to determine whether somatic mutation of any other oncogenes contribute to the development of lesions. In a second Aim we will use single-cell genomic DNA sequencing to determine whether individual endothelial cells require both the somatic CCM mutation and the PIK3CA mutation (and/or mutation of other genes discovered in Aim 1), or instead whether these genes are mutated in separate endothelial cell compartments of the lesion. In Aim 3, due to growing evidence that the somatically mutated cells poison the non-mutant cells to recruit them into the growing CCM, we will employ single-cell RNA sequencing to determine how the somatic mutation profile of the lesional cell influences the cell’s gene expression profile. In addition to increasing fundamental understanding of CCM pathogenesis, this work has therapeutic implications. The PI3K-AKT signaling pathway is a target of existing drugs with others under development. Thus as part of this project we will continue our quest for a highly effective therapy by testing compounds nominated in this or any of the three projects, using our more clinically-relevant CCM mouse models developed in the past funding cycle. With a more complete knowledge of the molecular and mutational signature of CCM lesional cells, we hope to identify new targets for drug repurposing for CCM patients.

摘要 - 项目 1 基于我们上一周期项目的发现，项目1将深入探讨 CCM 发病机制的细胞和分子事件。使用小鼠 CCM 病变的谱系追踪 我们表明，病变发生的初始事件是剩余野生型拷贝的体细胞丢失 Ccm 基因，随后进行具有相同体细胞突变的内皮细胞的克隆扩增，以及 似乎正在将额外的 EC 募集到不断生长的病变中——不含有 体细胞突变。其次，基于 Kahn 实验室关于激活 PI3 激酶在 在小鼠 CCM 模型中的病变发展中，我们发现超过 50% 的 PIK3CA 激活突变 人类 CCM 病变。这些合作发现表明我们尚未完全理解 有助于 CCM 病变发生、生长和成熟的分子和细胞事件。在第一个 目标是，我们将对大量人类 CCM 进行深度测序，以确定是否存在任何其他体细胞突变 癌基因有助于病变的发展。在第二个目标中，我们将使用单细胞基因组 DNA 测序以确定单个内皮细胞是否需要体细胞 CCM 突变 和 PIK3CA 突变（和/或目标 1 中发现的其他基因的突变），或者是否这些 基因在病变的不同内皮细胞区室中发生突变。在目标 3 中，由于增长 有证据表明，体细胞突变细胞会毒害非突变细胞，以将它们招募到生长中 CCM，我们将采用单细胞 RNA 测序来确定 病变细胞影响细胞的基因表达谱。除了增加基础 了解 CCM 发病机制，这项工作具有治疗意义。 PI3K-AKT 信号传导 途径是现有药物和其他正在开发的药物的目标。因此，作为该项目的一部分，我们将 通过测试本项或任何一项中提名的化合物，继续我们对高效疗法的探索 三个项目，使用我们在过去的资助周期中开发的更具临床相关性的 CCM 小鼠模型。 随着对 CCM 病变细胞的分子和突变特征有了更全面的了解，我们 希望为 CCM 患者确定药物再利用的新靶点。