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: 10621249
• 负责人: Douglas A. Marchuk
• 金额: $ 41.54万
• 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/10621249
• 关键词: AKT Signaling Pathway; Blood Vessels; Brain; Cell Compartmentation; Cell Lineage; Cell Separation; 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; Hemorrhage; Human; Individual; Knowledge; Laboratories; Learning; Lesion; Malignant Neoplasms; Modeling; Molecular; Mus; Mutate; Mutation; Normal Cell; Oncogenes; Operative Surgical Procedures; PIK3CA gene; PIK3CG 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; 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 根据我们计划项目上一个周期中的发现，项目1将深入研究 使用鼠ccm腿的谱系跟踪 我们表明，病变的初始事件是剩余的野生型副本的体细胞丢失 CCM基因，然后是具有相同体细胞突变的内皮细胞的克隆膨胀，并且 似乎将其他EC招募到成长中的病变中 - 不藏有的细胞 躯体突变。其次，基于Kahn Lab的发现，用于激活PI3激酶的角色 小鼠CCM模型中的病变发展，我们确定了超过50％的激活PIK3CA突变 人类CCM病变。这些协作发现表明我们尚未完全了解 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 目的，我们将深入序列人类CCM，以确定其他任何其他人的体细胞突变是否 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 以及PIK3CA突变（在AIM 1中发现的其他基因的突变），或者是否是这些 在病变的单独内皮细胞室中突变基因。在AIM 3中，由于成长 证据表明，体形突变的细胞毒害了非突变细胞以将其招募到增长中 CCM，我们将采用单细胞RNA测序来确定如何的体细胞突变曲线 病变细胞影响细胞的基因表达谱。除了增加基本 了解CCM发病机理，这项工作具有治疗意义。 PI3K-AKT信号传导 途径是与正在开发的其他人的现有药物的目标。作为这个项目的一部分，我们将 通过测试在此或任何一个中提名的化合物，继续我们寻求高效疗法 三个项目，使用我们在过去的融资周期中开发的更临床的CCM鼠标模型。 有了更完整的了解CCM病变细胞的分子和突变特征，我们 希望确定针对CCM患者的药物重新利用的新靶标。