MiR-152/PKM2/SLC7A5 axis in breast cancer development, chemo- and radiation-treatment response

MiR-152/PKM2/SLC7A5 轴在乳腺癌发展、化疗和放疗反应中的作用

• 批准号: 10445658
• 负责人: Jun He
• 金额: $ 64.48万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10445658
• 关键词: Affect; Alternative Splicing; Amino Acids; Animal Model; Biology; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer Treatment; Cell Line; Cell model; Cells; Chemotherapy and/or radiation; DNA; DNA Methylation; Data; Development; Doxorubicin; Endothelial Cells; Estrogen receptor positive; Family; Future; Goals; HNRPA3 gene; Heterogeneous-Nuclear Ribonucleoproteins; Homologous Gene; Human; Hypermethylation; IL8 gene; Malignant Neoplasms; Mediating; Metabolic; Methylation; MicroRNAs; Modeling; Molecular; Muscle; Pathway interactions; Play; Polypyrimidine Tract-Binding Protein; Promoter Regions; Protein Isoforms; Protein Kinase; Proteins; Pyruvate Kinase; Radiation; Radiation therapy; Research Project Grants; Resistance; Role; Signal Pathway; Testing; Therapeutic; Therapeutic procedure; Tissues; Tumor Angiogenesis; Tumor Tissue; Up-Regulation; Warburg Effect; Woman; angiogenesis; base; cancer cell; cancer survival; cancer therapy; chemoradiation; cohort; experimental study; improved; knock-down; malignant breast neoplasm; member; mortality; multidisciplinary; new therapeutic target; orthotopic breast cancer; overexpression; paracrine; patient derived xenograft model; receptor; solute; therapy resistant; translational potential; treatment response; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor metabolism; Affect; Alternative Splicing; Amino Acids; Animal Model; Biology; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer Treatment; Cell Line; Cell model; Cells; Chemotherapy and/or radiation; DNA; DNA Methylation; Data; Development; Doxorubicin; Endothelial Cells; Estrogen receptor positive; Family; Future; Goals; HNRPA3 gene; Heterogeneous-Nuclear Ribonucleoproteins; Homologous Gene; Human; Hypermethylation; IL8 gene; Malignant Neoplasms; Mediating; Metabolic; Methylation; MicroRNAs; Modeling; Molecular; Muscle; Pathway interactions; Play; Polypyrimidine Tract-Binding Protein; Promoter Regions; Protein Isoforms; Protein Kinase; Proteins; Pyruvate Kinase; Radiation; Radiation therapy; Research Project Grants; Resistance; Role; Signal Pathway; Testing; Therapeutic; Therapeutic procedure; Tissues; Tumor Angiogenesis; Tumor Tissue; Up-Regulation; Warburg Effect; Woman; angiogenesis; base; cancer cell; cancer survival; cancer therapy; chemoradiation; cohort; experimental study; improved; knock-down; malignant breast neoplasm; member; mortality; multidisciplinary; new therapeutic target; orthotopic breast cancer; overexpression; paracrine; patient derived xenograft model; receptor; solute; therapy resistant; translational potential; treatment response; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor metabolism

Breast cancer (BCa) is the leading cause of women cancer mortality worldwide, and triple negative breast cancer (TNBC) cells accounts for 15-20% BCa. However, higher therapeutic resistance and lower survival of TNBC patients remain the major hinder of BCa treatment. Doxorubicin (Dox) and radiation are commonly used for TNBC treatment. In our preliminary study, we found that TNBC showed much lower expression levels of miR-152. Higher expression levels of zeste homologue 2 (EZH2) and DNA hypermethylation were involved in miR-152 suppression in TNBC cells. To understand mechanism of miR-152 suppression, and role and mechanism of miR-152 in regulating cancer development and Dox- and radiation-induced resistance, we performed a lot of preliminary experiments to identify what molecules that were potential miR-152 direct targets as well as that were upregulated in Dox-resistant TNBC or PDX tumor tissues. We found that pyruvate kinase muscle 2 (PKM2), solute Carrier Family 7 Member 5 (SLC7A5), and polypyrimidine tract-binding protein (PTBP1) as direct targets of miR-152 with potential ability to converse Dox-mediated resistance. PKM2, SLC7A5 and PTBP1 levels were increased when miR-152 expression was suppressed in TNBC cells and PDX model. We found that miR-152 suppression played an important role in mediating TNBC metabolic reprogramming, Warburg effect switch, tumor growth and Dox resistance through PKM2/SLC7A5/PTBP1. We showed that overexpression of miR-152 or PKM2 knockdown rendered TNBC cells more sensitive to radiation treatment. These new findings create new opportunity to investigate mechanism of TNBC therapeutic resistance. We hypothesize that resistance to chemotherapy and radiation treatment, metabolic reprogramming, and TNBC development are induced by miR-152 suppression and upregulation of PKM2, SLC7A5, and PTBP1. To test this central hypothesis, we will perform experiments through three aims. In Aim 1, we will investigate role and mechanism of miR-152 suppression in TNBC cells in doxorubicin- and radiation-induced resistance; and role and mechanism of PKM2 switch and induction via alternative splicing by miR-152 suppression to induce Warburg effect, therapeutic resistance, and tumor growth. In Aim 2, we will determine role and mechanism of miR- 152/SLC7A5/PTBP1 pathway in regulating metabolic reprogramming and Warburg effect switch, and in mediating doxorubicin- and radiation-induced therapeutic resistance. In Aim 3, we will determine whether miR-152 suppression and PKM2 induction regulate tumor angiogenesis through CXCL8 expression using a humanized chimeric tumor model; and whether levels of miR-152, PKM2, HIF-1α, hnRNPA3, SLC7A5, and PTBP1 are correlated each other, and are correlated with therapeutic responses and with the cancer stages and survival. This R01 project will identify mechanisms of therapeutic resistance, and metabolic reprogramming; and provide information for developing new treatment option of TNBC in the future.

乳腺癌（BCA）是全世界女性癌症死亡率的主要原因，而三重阴性乳房 癌症（TNBC）细胞占BCA的15-20％。但是，较高的治疗性耐药性和较低的存活率 TNBC患者中的主要阻碍是BCA治疗。阿霉素（DOX）和辐射通常是 用于TNBC治疗。在我们的初步研究中，我们发现TNBC的表达低得多 miR-152的水平。 Zeste同源2（EZH2）和DNA高甲基化的较高表达水平为 参与TNBC细胞中的miR-152抑制。了解miR-152抑制的机制，以及 miR-152在调节癌症发展和DOX和辐射诱导的作用和机制 抗性，我们进行了许多初步实验，以识别哪些潜在分子 miR-152直接靶标以及在耐DOX的TNBC或PDX肿瘤组织中进行了更新。我们 发现丙酮酸激酶2（PKM2），可溶性载体家族7成员5（SLC7A5）和 息肉嘧啶片结合蛋白（PTBP1）作为MiR-152的直接靶标，具有转换的潜在能力 DOX介导的电阻。当miR-152表达时，PKM2，SLC7A5和PTBP1水平升高 在TNBC细胞和PDX模型中被抑制。我们发现miR-152抑制作用起很重要 在介导TNBC代谢重编程，Warburg效应开关，肿瘤生长和DOX抗性中的作用 通过PKM2/SLC7A5/PTBP1。我们表明miR-152或PKM2敲低的过表达呈现 TNBC细胞对放射治疗更敏感。这些新发现为调查创造了新的机会 TNBC治疗耐药性的机理。我们假设对化学疗法的抵抗力和 辐射处理，代谢重编程和TNBC发育由miR-152诱导 PKM2，SLC7A5和PTBP1的抑制和上调。为了检验这个中心假设，我们将 通过三个目标执行实验。在AIM 1中，我们将研究mir-152的作用和机制 阿霉素和辐射诱导的抗性中TNBC细胞抑制；以及角色和机制 PKM2通过miR-152抑制通过替代剪接而诱导和诱导，以影响Warburg效应， 治疗性抗性和肿瘤生长。在AIM 2中，我们将确定mir的作用和机制 152/slc7a5/ptbp1途径在调节代谢重编程和沃伯格效应开关中以及 介导阿霉素和辐射诱导的治疗性抗性。在AIM 3中，我们将确定是否 miR-152抑制和PKM2诱导通过CXCL8表达调节肿瘤血管生成 人构化嵌合肿瘤模型；以及miR-152，PKM2，HIF-1α，HNRNPA3，SLC7A5和 PTBP1相互关联，与治疗反应相关，与癌症阶段相关 和生存。这个R01项目将确定热阻力和代谢的机制 重编程；并为将来开发TNBC的新治疗选择提供信息。