The PLOD2/succinate axis in cancer cell plasticity and stemness

PLOD2/琥珀酸轴在癌细胞可塑性和干细胞性中的作用

• 批准号: 10680932
• 负责人: Ren Xu
• 金额: $ 36.57万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10680932
• 关键词: Aftercare; Biochemistry; Biological; Biometry; Breast; Breast Cancer Cell; Breast Epithelial Cells; Breast cancer metastasis; Cell Membrane Permeability; Cell physiology; Cells; ChIP-seq; Chromatin; Citric Acid Cycle; Clinical; Collagen; CpG Islands; Cytoplasm; DDR1 gene; DNA; Development; Dioxygenases; Endoplasmic Reticulum; Enzymes; Epigenetic Process; Exhibits; FDA approved; Fibroblasts; Fibrosis; Gene Expression Profiling; Genetic Transcription; Human; Hydroxylation; In Vitro; Incidence; Knockout Mice; Knowledge; Loperamide; Lysine; Malignant Neoplasms; Mediating; Mesenchymal; Metabolic; Modeling; Modification; Molecular; Mouse Mammary Tumor Virus; Mus; Neoplasm Metastasis; Pathologist; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Positioning Attribute; Procollagen; Prognosis; Regulation; Relapse; Repression; Research; Role; Scheme; Succinates; TP53 gene; Testing; Tissue Sample; Tissues; Weight; Xenograft Model; alpha ketoglutarate; breast cancer progression; cancer cell; cancer subtypes; cell growth regulation; conditional knockout; crosslink; epigenetic regulation; epithelial to mesenchymal transition; experimental study; gain of function; inhibitor; loss of function; malignant breast neoplasm; metabolomics; mid-career faculty; mouse model; multidisciplinary; neoplastic cell; novel; patient derived xenograft model; screening; small hairpin RNA; stable isotope; stemness; therapeutic target; transcription factor; transcriptome sequencing; triple-negative invasive breast carcinoma

Abstract Tumor cells in TNBC tissue exhibit enhanced cell plasticity and stemness, which is crucial for cancer metastasis. Metabolic and epigenetic reprogramming plays crucial roles in the regulation of cell plasticity. Succinate is an intermediate metabolite of the tricarboxylic acid (TCA) cycle. Aberrant accumulation of succinate has been detected in many cancers. However, the cellular function and regulation of succinate in breast cancer development and progression have not been well investigated. We showed that the epithelial-to-mesenchymal transition (EMT) was associated with profound changes in metabolites, including elevation of cytoplasmic succinate. Importantly, treatment with the membrane permeable succinate was sufficient to induce mesenchymal phenotypes, enhance cancer cell stemness and colonization, and reduce DNA hydroxymethylation and gene transcription in mammary epithelial cells. Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase 2 (PLOD2) catalyzes lysine hydroxylation in endoplasmic reticulum and generates succinate as another product. We found that PLOD2 expression was induced in cancer cells in TNBC tissue, and increased PLOD2 expression correlates with poor prognosis and high chances of cancer metastasis. Silence of PLOD2 in cancer cells significant reduced cancer cell colonization and metastasis. PLOD2 expression was induced during EMT and associated with elevation of cytoplasmic succinate levels. Using gain- and loss-of function approaches, we showed that increased PLOD2 expression is necessary and sufficient to enhance the accumulation of cytoplasmic succinate. Importantly, silencing PLOD2 in breast cancer cells inhibited cancer cell stemness. The overall objective of this project is to define roles of the PLOD2/succinate axis in regulating cancer cell plasticity and stemness, and to evaluate biological activity of the new PLOD2 inhibitor in suppressing TNBC progression and metastasis. The central hypothesis of this proposal is that PLOD2-induced succinate accumulation enhances cancer cell plasticity and metastasis by reducing DNA hydroxymethylation; therefore, targeting the PLOD2/succinate axis is a potential strategy to halt TNBC progression and metastasis. To test this hypothesis, we propose the following aims: Aim 1. Elucidate the molecular and cellular mechanisms by which succinate enhances cancer cell plasticity and stemness. Aim 2. Define roles of the PLOD2/succinate axis in regulating breast cancer progression. Aim 3. Evaluate the potential of targeting PLOD2 to suppress breast cancer metastasis. Successfully completion of the project will reveal novel function of collagen hydroxylation enzymes in regulating succinate-associated epigenetic reprogramming and identify PLOD2 as a potential therapeutic target to suppress breast cancer metastasis.

抽象的 TNBC组织中的肿瘤细胞表现出增强的细胞塑性和干性，这对于癌症转移至关重要。 代谢和表观遗传重编程在细胞可塑性的调节中起着至关重要的作用。琥珀酸盐是一个 三羧酸（TCA）周期的中间代谢产物。琥珀酸酯的异常积累 在许多癌症中检测到。但是，乳腺癌中琥珀酸酯的细胞功能和调节 发展和进步尚未得到很好的研究。我们表明上皮到间质 过渡（EMT）与代谢产物的深刻变化有关，包括细胞质升高 琥珀酸酯。重要的是，用膜可渗透琥珀酸酯的治疗足以诱导间质 表型，增强癌细胞的干性和定殖，并降低DNA羟甲基和基因 乳腺上皮细胞中的转录。 Procollagen-赖氨酸，2-氧化5-氧化酶2（PLOD2） 催化内质网中赖氨酸羟基化，并产生琥珀酸酯作为另一种产品。我们发现 在TNBC组织中的癌细胞中诱导了PLOD2的表达，而PLOD2表达的增加与 预后不良，癌症转移的机会很高。癌细胞中PLOD2的静音显着降低 癌细胞定植和转移。在EMT期间诱导PLOD2的表达，并与 胞质琥珀酸盐水平的升高。使用功能丧失方法，我们表明 PLOD2表达增加是必要的，足以增强细胞质琥珀酸酯的积累。 重要的是，使乳腺癌细胞中的PLOD2沉默会抑制癌细胞的干性。总体目标 项目是定义PLOD2/琥珀酸酯轴在调节癌细胞的可塑性和干性中的作用，并 评估新的PLOD2抑制剂在抑制TNBC进展和转移时的生物学活性。这 该提议的中心假设是PLOD2诱导的琥珀酸酯积累增强了癌细胞 可塑性和转移通过还原DNA羟甲基化；因此，靶向PLOD2/琥珀牙轴为 阻止TNBC进展和转移的潜在策略。为了检验这一假设，我们提出以下 目的：目标1。阐明琥珀酸增强癌细胞可塑性的分子和细胞机制 和茎。 AIM 2。定义PLOD2/琥珀酸轴在调节乳腺癌进展中的作用。目标3。 评估靶向PLOD2抑制乳腺癌转移的潜力。成功完成 项目将揭示胶原蛋白羟基化酶在调节与琥珀酸盐相关的表观遗传学中的新功能 重新编程并识别PLOD2是抑制乳腺癌转移的潜在治疗靶标。