Clonogenic Characterization of Myeloma Progenitor Cells

骨髓瘤祖细胞的克隆形成特征

• 批准号: 8104170
• 负责人: LIA ELENA PEREZ
• 金额: $ 17.62万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-02 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8104170
• 关键词: Antibodies; Apoptotic; Applications Grants; B lymphoid malignancy; B-Cell Development; B-Lymphocytes; Biological Assay; Biology; Bone Marrow; Bortezomib; Cancer Biology; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cell physiology; Cells; Clinical; Clinical Trials; Clonality; Clone Cells; Combination Drug Therapy; Coupled; DNA Sequence Rearrangement; Development; Disease; Disease-Free Survival; Drug resistance; Elements; Environment; Fanconi' s Anemia; Feedback; Fibronectins; Future; Gene Targeting; Generations; Goals; Growth; Heavy-Chain Immunoglobulins; Hematologic Neoplasms; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Immunotherapy; In Vitro; Label; Ligands; Liquid Chromatography; Maintenance; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mediator of activation protein; Multiple Myeloma; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Plant Roots; Plasma Cells; Play; Population; Property; Proteins; Recurrence; Recurrent disease; Regulation; Relapse; Residual Neoplasm; Resistance; Role; Signal Pathway; Signal Transduction; Staging; Stem cells; Testing; Treatment Efficacy; Treatment Failure; Up-Regulation; base; cancer cell; cancer stem cell; cell stroma; chemotherapy; clinically significant; cytokine; improved; in vivo; liquid chromatography mass spectrometry; multiple reaction monitoring; novel; prevent; progenitor; protein expression; prototype; public health relevance; resistance mechanism; response; self-renewal; syndecan; tool; treatment strategy; tumor

DESCRIPTION (provided by applicant): Multiple Myeloma (MM) is the prototype of a clonal B-cell malignancy. Hematopoietic stem cell (HSC) transplantation and/or novel chemotherapy combinations have improved long-term disease free survival. However, relapse of the underlying disease remains the primary cause of treatment failure with MM being essentially an incurable disease. Therefore, strategies aiming to improve treatment efficacy are much needed. The central hypothesis of this project is that bone marrow sub-populations in MM patients constitute a progenitor compartment with self renewal capacity that is responsible for disease initiation, maintenance and progression. These cells may have a specific signature to render them insensitive to chemotherapy and they may rely on the hematopoietic stroma microenvironment for their survival. Our preliminary results show that MM patient's bone marrow contain a clonal sub-population that do not express syndecan-1 (CD138-), differentiates in clonogenic assay to a mature cell (CD138+) and is relatively resistant to bortezomib treatment. The proposed study will contribute to the understanding of MM biology analyzing the phenotypic and unique make up of a MM progenitor sub-population that may be in part responsible for disease initiation and/or relapse. Our future goal is to establish the clinical significance of these cells and to develop drugs and/or immune therapies to target all MM sub-populations. In Aim 1, we will determine the differentiation potential and self-renewal capacity of CD138- MM cells. We proposed to characterize phenotypic differentiation of CD138- MM cells and establish their chromosomal signature. We plan to define their clonality with patient-specific immunoglobulin heavy chain rearrangements and clonogenicity in vivo. In Aim 2, we will determine the role of self renewal pathways as an intrinsic mechanism of resistance of CD138- MM sub-populations by analyzing related proteins by mass spectrometry. We will test the effect of self renewal ligands on protein expression, on cell proliferation, on activation of target genes and clonogenic growth of CD138- cells. In Aim 3, we will determine the role of stroma cells in conferring a survival advantage to CD138-cells. I hypothesize that the microenvironment supports CD138- MM cells clonogenicity and these cells depend on it signals for their survival as the stroma provides a supporting niche. I postulate that stroma elements may provide cyto-protection to CD138- MM subpopulations in part due to perturbations in cell cycle progression and/or known MM-stroma survival mechanism. In pursuit of this aim we will assess stroma related cytokines and fibronectin effect on cell cycle and on modulation of protein expression of BcL-2 apoptotic regulators by mass spectrometry and correlation with drug response. PUBLIC HEALTH RELEVANCE: Multiple Myeloma (MM) is an incurable hematological malignancy of plasma cells. Plasma cells grow in the bone marrow (BM) and produce same type of abnormal antibodies (clone). Development of strategies to improve treatment efficacy and to prevent recurrence of disease (relapse) are much needed. An important cause of treatment failure in cancer is drug resistance leading to minimal residual disease (MRD). Developing novel treatment strategies to target mediators of MRD is essential for complete disease eradication. The "cancer stem cell" concept debates that some cells within the tumor are the root of cancer by producing cancer cells and cancer cure depends on the eradication of these cells. To date, the clinical role of these cells is cancer biology is unknown and deep understanding of these cells in MM remains to be determined. Our preliminary results show that MM BM contains a clone of cells that do not express markers used to identify mature MM cells (plasma cells) and are less susceptible to chemotherapy. We hypothesize that these cells maybe responsible for the origin of MM and responsible for MRD. We propose to establish whether these cells play a role in MM generation and to understand growth regulation mechanisms of these cells. We plan to study self renewal pathways and interaction with the BM micro-environment since we believe that this niche provides positive feedback for their survival. Understanding the role of these cell in MM and decipher their survival tools will be essential since eradication of these cells should produce curative results in MM if hypothesis is proven.

描述(申请人提供)：多发性骨髓瘤(MM)是一种克隆性B细胞恶性肿瘤的原型。造血干细胞(HSC)移植和/或新的化疗组合提高了长期无病生存率。然而，基础疾病的复发仍然是治疗失败的主要原因，多发性骨髓瘤基本上是一种不治之症。因此，迫切需要旨在提高治疗效果的策略。这个项目的中心假设是MM患者的骨髓亚群构成了一个具有自我更新能力的祖细胞亚群，负责疾病的发生、维持和发展。这些细胞可能有一个特殊的特征，使它们对化疗不敏感，它们可能依靠造血基质微环境生存。我们的初步结果表明，MM患者的骨髓含有一个不表达Syndecan-1(CD138-)的克隆性亚群，在克隆形成实验中分化为成熟细胞(CD138+)，并且对Bortezomib治疗相对耐药。这项拟议的研究将有助于理解多发性骨髓瘤生物学，分析多发性骨髓瘤前体亚群的表型和独特构成，这可能在一定程度上导致疾病的起始和/或复发。我们未来的目标是确定这些细胞的临床意义，并开发针对所有MM亚群的药物和/或免疫疗法。在目标1中，我们将测定CD138-MM细胞的分化潜能和自我更新能力。我们建议鉴定CD138-MM细胞的表型分化，并建立它们的染色体特征。我们计划用患者特异性免疫球蛋白重链重排和体内克隆性来确定它们的克隆性。在目标2中，我们将通过质谱分析相关蛋白质来确定自我更新途径作为CD138-MM亚群耐药的内在机制。我们将测试自我更新配体对CD138-细胞的蛋白质表达、细胞增殖、靶基因激活和克隆性生长的影响。在目标3中，我们将确定基质细胞在赋予CD138-细胞生存优势中的作用。我假设微环境支持CD138-MM细胞的克隆性，这些细胞依靠微环境信号生存，因为基质提供了支持的利基。我推测，间质成分可能对CD138-MM亚群提供细胞保护，部分原因是细胞周期进程和/或已知的MM-间质生存机制的扰动。为了达到这一目的，我们将通过质谱学方法评估基质相关细胞因子和纤维连接蛋白对细胞周期的影响，以及对bcl2凋亡调节蛋白表达的调节作用，并与药物反应的相关性进行评估。 公共卫生相关性：多发性骨髓瘤(MM)是一种无法治愈的浆细胞恶性血液病。浆细胞在骨髓(BM)中生长，并产生相同类型的异常抗体(克隆)。迫切需要制定提高治疗效果和防止疾病(复发)复发的战略。癌症治疗失败的一个重要原因是导致微小残留病(MRD)的耐药性。开发针对MRD介体的新治疗策略对于彻底根除疾病至关重要。“癌症干细胞”的概念认为，肿瘤内的一些细胞通过产生癌细胞而成为癌症的根源，癌症的治疗取决于这些细胞的根除。到目前为止，这些细胞在肿瘤生物学中的临床作用尚不清楚，对这些细胞在多发性骨髓瘤中的深入了解仍有待确定。我们的初步结果表明，MM BM含有克隆的细胞，不表达用于识别成熟MM细胞(浆细胞)的标记，并且对化疗不太敏感。我们推测，这些细胞可能与MM的起源有关，也可能与MRD有关。我们建议确定这些细胞是否在多发性骨髓瘤的发生中发挥作用，并了解这些细胞的生长调节机制。我们计划研究自我更新的途径以及与BM微环境的相互作用，因为我们相信这个利基为它们的生存提供了积极的反馈。了解这些细胞在多发性骨髓瘤中的作用并破译它们的生存工具将是至关重要的，因为如果假设得到证实，根除这些细胞应该会在多发性骨髓瘤中产生疗效。

项目成果

Stemness of B-cell progenitors in multiple myeloma bone marrow.

多发性骨髓瘤骨髓中B细胞祖细胞的干性。

• DOI: 10.1158/1078-0432.ccr-12-0531
• 发表时间: 2012-11-15
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Boucher K;Parquet N;Widen R;Shain K;Baz R;Alsina M;Koomen J;Anasetti C;Dalton W;Perez LE
• 通讯作者: Perez LE