(PQ1) Genomic characterization of mesenchymal stromal cells in Monoclonal Gammopathy of Undermined Significance (MGUS)

(PQ1) 意义被削弱的单克隆丙种球蛋白病 (MGUS) 中间充质基质细胞的基因组特征

• 批准号: 9101485
• 负责人: Irene M. Ghobrial
• 金额: $ 43.85万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9101485
• 关键词: 1p13; Affect; Age-Years; Anemia; Applications Grants; Bone Marrow; CRISPR/Cas technology; Cell Aging; Cells; Chromosomal translocation; Chromosome abnormality; Clonal Evolution; Clonal Expansion; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Copy Number Polymorphism; DNA Sequence Alteration; Development; Diagnostic; Disease; Disease Progression; Distant; Event; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genomics; Grant; Hypercalcemia; Image; Immunoglobulin A; Immunoglobulin G; In Vitro; Kidney Failure; Lead; Lesion; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mesenchymal; Modeling; Molecular Abnormality; Monoclonal Gammapathies; Monoclonal gammopathy of uncertain significance; Multiple Myeloma; Mus; Mutation; NRAS gene; Participant; Patients; Plasma Cells; Point Mutation; Population; Premalignant; Proliferating; Property; Proteins; Regulation; Risk; Risk Reduction; Role; Sampling; Serum; Site; Somatic Mutation; Staging; Stem cells; Stromal Cells; Symptoms; Testing; Therapeutic Agents; Therapeutic Intervention; Tissues; Validation; base; bone; cancer cell; cell age; cell type; chromosome loss; design; epigenomics; in vivo; in vivo Model; knockout gene; mouse model; multiple myeloma M Protein; novel; prevent; public health relevance; research study; transcriptomics; tumor; tumor initiation; tumor progression; tumorigenesis

 DESCRIPTION (provided by applicant): Monoclonal gammopathy of undetermined significance (MGUS) is one of the most common pre-malignant disorders and affects approximately 3.5% of the population over 50 years of age. This grant application aims to test the provocative question PQ1. For tumors that arise from a pre-malignant field, what properties of cells in this field can be used to design strategies to inhibit the development of future tumors Recent studies showed that tumors are more than insular masses of proliferating cancer cells. Instead, they are complex tissues composed of multiple distinct cell types that participate in heterotypic interactions with one another. Here, we hypothesize that normal bone marrow mesenchymal stromal cells (MSCs) adjacent to the early premalignant MGUS cells are active participants in tumorigenesis and clonal evolution rather than passive bystanders; as such, these cells contribute to the development of multiple myeloma (MM). We will test this hypothesis in 3 specific Aims. In Specific Aim 1, we will elucidate genomic/transcriptomic events that govern alterations in MSCs at the premalignant stage of MGUS. We will perform integrative characterization of genomic, epigenomic and transcriptomic changes that occur in MSCs present near MGUS cells in bulk and at the single cell level. Further validation of specific target will be performed at the protein level to identify the spatial localization of these altered MSCs i the bone marrow niche using CyTOF mass spectrometry imaging. In Specific Aim 2, we will identify the sequential acquisition of genomic lesions in MSCs in the early premalignant stage and during disease progression in a murine model. Here, we will define the changes that occur in MSCs during the first genetic event in the premalignant plasma cells and identify changes that occur in the proximity of clonal plasma cells at the early MGUS stages vs. those that are present in distant bone marrow sites. We will examine whether MSCs alterations precede the early genetic hit (permissive microenvironment) or are acquired after MGUS development (acquired alterations). We will also define genomic and gene expression changes that occur in MSCs with stem cell aging in these mice. In Specific Aim 3, we will validate specific targets using CRISPR-based gene knockout of highly prioritized genes based on the studies performed in Aims 1 and 2 to functionally interrogate their specific role on MSCs and their regulation of tumor progression using in vitro and in vivo models. Furthermore, we will develop MSC-specific genomic alterations using CRISPR-cas9 mice crossed with Osx-cre mice to define the role of these target genes in regulating tumor initiation and MGUS progression in murine MM models. These focused research studies will help define the contributing role of MSCs in the early stages of MGUS development and clonal progression to MM. By identifying novel targets that regulate clonal evolution at the early premalignant stage of MGUS, we may be able to develop therapeutic agents that prevent or delay progression from MGUS to overt MM. Indeed, by eradicating the disease at the precursor stages, MM may become a preventable disease.

 描述（由申请人提供）：意义不明的单克隆丙种球蛋白病 (MGUS) 是最常见的癌前病变之一，影响约 3.5% 的 50 岁以上人口。本拨款申请旨在测试具有争议性的问题 PQ1。对于起源于恶变前区域的肿瘤，可以利用该区域细胞的哪些特性来设计抑制未来肿瘤发展的策略。最近的研究表明，肿瘤更容易发生。相反，它们是由多种不同细胞类型组成的复杂组织，这些细胞类型彼此之间存在异型相互作用。在这里，我们捕获了与早期癌前 MGUS 细胞相邻的正常骨​​髓间充质基质细胞 (MSC)。是肿瘤发生和克隆进化的积极参与者，而不是被动的旁观者；因此，这些细胞有助于多发性骨髓瘤 (MM) 的发展。具体目标 1，我们将阐明在 MGUS 癌前阶段控制 MSC 变化的基因组/转录组事件。我们将对 MGUS 细胞附近和单个细胞中存在的 MSC 中发生的基因组、表观基因组和转录组变化进行综合表征。将在蛋白质水平上对特定目标进行进一步验证，以使用 CyTOF 质谱成像来识别这些改变的 MSC 在骨髓生态位中的空间定位。在具体目标 2 中，我们将在小鼠模型中确定癌前阶段和疾病进展期间 MSC 中病变基因组的顺序获取。在这里，我们将定义癌前血浆中第一个遗传事件期间 MSC 中发生的变化。细胞并识别早期 MGUS 阶段克隆浆细胞附近发生的变化与远处骨髓部位存在的变化，我们将检查 MSC 的改变是否先于早期基因打击（允许的微环境）。或在 MGUS 发育后获得（获得性改变）。我们还将定义这些小鼠干细胞老化时 MSC 中发生的基因组和基因表达变化。在特定目标 3 中，我们将使用基于 CRISPR 的高度基因敲除来验证特定目标。根据目标 1 和 2 中进行的研究对基因进行优先排序，以利用体外和体内模型在功能上探讨它们对 MSC 的特定作用及其对肿瘤进展的调节。此外，我们将使用开发 MSC 特异性基因组改变。 CRISPR-cas9 小鼠与 Osx-cre 小鼠杂交，以确定这些靶基因在调节小鼠 MM 模型中肿瘤发生和 MGUS 进展中的作用。这些重点研究将有助于确定 MSC 在 MGUS 发育和早期阶段的贡献作用。通过确定在 MGUS 早期癌前阶段调节克隆进化的新靶标，我们或许能够开发出预防或延缓从 MGUS 进展为明显 MM 的治疗药物。在前兆阶段根除该疾病，MM 可能成为一种可预防的疾病。