Role of an Aberrant N6-Methyladenosine-LncRNA Axis in the Development and Maintenance of Drug Resistance through Regulating the Leukemia Stem Cell

异常的 N6-甲基腺苷-LncRNA 轴在通过调节白血病干细胞产生和维持耐药性中的作用

• 批准号: 10701762
• 负责人: Gang Huang
• 金额: $ 59.11万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701762
• 关键词: Address; Biological Markers; CD44 gene; Cancer Patient; Cell Survival; Cells; Critical Pathways; Development; Disease; Disease Progression; Disease remission; Drug resistance; Engraftment; Gene Expression; Genes; Genetic Transcription; Goals; IL2RA gene; Impairment; Knowledge; Leukemic Cell; Link; Malignant Neoplasms; Methylation; Modeling; Modification; Molecular; Mus; Outcome; Pathway interactions; Patients; Positioning Attribute; Process; Proliferating; Protein Tyrosine Kinase; Proto-Oncogene Protein c-kit; RNA; Recurrent disease; Refractory; Regimen; Regulation; Research; Resistance; Resistance development; Role; Signal Transduction; Source; Testing; Therapeutic; Translational Research; Treatment Efficacy; Treatment Failure; Tyrosine Kinase Inhibitor; Untranslated RNA; Up-Regulation; Work; advanced disease; cancer drug resistance; cancer stem cell; cell growth; clinical biomarkers; clinical translation; demethylation; design; differential expression; drug maintenance; drug sensitivity; epitranscriptomics; experience; fat mass and obesity-associated protein; gain of function; gene repression; improved; inhibitor; inhibitor therapy; innovation; knock-down; leukemia; leukemia treatment; leukemic stem cell; loss of function; methylome; molecular targeted therapies; mouse model; new therapeutic target; novel; novel strategies; patient response; pharmacologic; pre-clinical; refractory cancer; response; self-renewal; stem cell biomarkers; stem cell fate; stem cell function; stem cell self renewal; stemness; targeted treatment; treatment strategy; tumor progression

Tyrosine kinase targeted (TKI) therapies have revolutionized leukemia treatment, but TKIs are not able to kill leukemia stem cells (LSCs), which are responsible for propagating and disease recurrence, and believed to be the source of treatment failure. Our long-term goals are to further address how LSC persistence is regulated, and to develop new LSC-eliminating treatment strategies to improve cure rates and survival. The short-term goals of this research are to determine whether a N6-methyladenosine (m6A)--long non-coding RNA (lncRNA) axis regulates LSC stemness and persistence during TKI selection process, and to explore the therapeutic potential of targeting the m6A-lncRNA axis for eradicating TKI resistant LSCs and also decipher the underlying molecular mechanisms. The m6A methylation is the most common epitranscriptomic modification on RNAs (i.e., lncRNAs), and crucially regulates lncRNA-initiated gene expression. lncRNA abnormalities frequently associate with cancer disease progression and drug resistance. The preliminary evidence linking an m6A-lncRNA axis to resistant LSCs is from our proof of principle studies demonstrating that i) a dynamic and reversible m6A methylome determined by fat mass and obesity-associated protein (FTO) helps leukemia cells avoid TKI killing leading to TKI resistance; ii) there are many lncRNAs (annotated) that are differentially expressed in resistant versus sensitive cells. About 50% of these lncRNAs bear m6A motifs and have the changed m6A amounts in resistant cells, collectively, suggesting a unique lncRNA signature that is specific to TKI resistance and is regulated by m6A methylation; iii) upregulation of these m6A-associated lncRNAs in patients who do not respond to TKIs is predicative of worse outcomes, and knockdown of them impairs resistant cell growth and renders resistant cells sensitive to TKIs; iv) compared to sensitive ones, TKI-resistant cells highly express LSC markers (CD117, CD44, CD25, CD133) whose upregulation is associated with m6A reduction. Our hypothesis is that the FTO-m6A-lncRNA cascade may be a critical pathway to control LSC persistence to TKIs and a new druggable target to eradicate persistent LSCs improving TKI cure rates. We will test our hypothesis through three aims: 1) Determine how the FTO-m6A axis regulates lncRNA aberrations in TKI resistance; 2) Determine whether and how a dynamic m6A methylome regulates LSC persistence to TKIs; 3) Determine whether and how pharmacological targeting of the FTO-m6A-lncRNA cascade kills persistent LSCs using preclinical leukemia models. The proposed studies are conceptually innovative, because it targets a new pathway (the FTO-m6A- lncRNA cascade) in understanding LSC persistence to TKIs and in developing new regimens to eliminate TKI resistant LSCs. The proposed research is significant, because the findings will a) identify new pathways (i.e., FTO-m6A-lncRNA cascade) that regulate LSC persistence, deepening the molecular understanding of LSCs and lncRNA functions; b) develop new approaches (targeting the FTO-m6A-lncRNA cascade) to eliminate persistent LSCs improving the management of patients with refractory leukemia.

酪氨酸激酶靶向 (TKI) 疗法彻底改变了白血病治疗，但 TKI 无法杀死癌细胞 白血病干细胞（LSC）负责增殖和疾病复发，被认为是 治疗失败的根源。我们的长期目标是进一步解决 LSC 持久性的监管问题， 并开发新的 LSC 消除治疗策略以提高治愈率和生存率。短期 本研究的目标是确定 N6-甲基腺苷 (m6A)——长非编码 RNA (lncRNA) 轴在 TKI 选择过程中调节 LSC 干性和持久性，并探索治疗方法 靶向 m6A-lncRNA 轴消除 TKI 耐药性 LSC 的潜力，并破译其潜在机制 分子机制。 m6A 甲基化是 RNA 上最常见的表观转录组修饰（即 lncRNA），并且关键地调节lncRNA启动的基因表达。 lncRNA异常经常与 与癌症疾病进展和耐药性有关。 m6A-lncRNA 轴与 抗性 LSC 来自我们的原理研究证明，证明 i) 动态且可逆的 m6A 由脂肪量和肥胖相关蛋白 (FTO) 确定的甲基化组有助于白血病细胞避免 TKI 杀死 导致 TKI 耐药； ii）有许多lncRNA（已注释）在耐药性中差异表达 与敏感细胞相比。这些 lncRNA 中大约 50% 带有 m6A 基序，并且 m6A 数量发生了变化 耐药细胞共同表明了一种独特的 lncRNA 特征，该特征是 TKI 耐药所特有的，并且是 受 m6A 甲基化调节； iii) 无反应患者中这些 m6A 相关 lncRNA 的上调 TKI 的使用预示着更糟糕的结果，并且它们的敲低会损害耐药细胞的生长并导致 对 TKI 敏感的耐药细胞； iv) 与敏感细胞相比，TKI 耐药细胞高度表达 LSC 标记 （CD117、CD44、CD25、CD133）其上调与 m6A 减少相关。我们的假设是 FTO-m6A-lncRNA 级联可能是控制 LSC 对 TKI 持久性的关键途径，也是一种新的药物 目标是根除持续性 LSC，提高 TKI 治愈率。我们将通过三个目标来检验我们的假设：1） 确定 FTO-m6A 轴如何调节 TKI 耐药中的 lncRNA 畸变； 2) 确定是否和 动态 m6A 甲基化组如何调节 LSC 对 TKI 的持久性； 3) 确定是否以及如何 FTO-m6A-lncRNA 级联的药理学靶向可利用临床前白血病杀死持续性 LSC 模型。拟议的研究在概念上具有创新性，因为它针对的是一条新途径（FTO-m6A- lncRNA 级联）了解 LSC 对 TKI 的持久性并开发消除 TKI 的新方案 耐药LSC。拟议的研究意义重大，因为研究结果将 a) 确定新的途径（即 FTO-m6A-lncRNA 级联）调节 LSC 持久性，加深对 LSC 的分子理解 lncRNA功能； b) 开发新方法（针对 FTO-m6A-lncRNA 级联）以消除持久性 LSC 改善难治性白血病患者的治疗。