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.

靶向酪氨酸激酶（TKI）疗法彻底改变了白血病治疗，但TKI无法杀死 白血病干细胞（LSC），负责传播和疾病复发，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持久性如何调节， 并制定新的LSC淘汰治疗策略，以提高治愈率和生存。短期 这项研究的目标是确定N6-甲基读腺苷（M6A）是否 - 长的非编码RNA（LNCRNA） 轴在TKI选择过程中调节LSC的茎和持久性，并探索治疗性 靶向M6A-LNCRNA轴以根除TKI抗TKI LSC，并破译基础 分子机制。 M6A甲基化是RNA上最常见的表面参考的修饰（即 lncRNA），并至关重要地调节lncRNA引起的基因表达。 LNCRNA异常经常关联 具有癌症疾病进展和耐药性。将M6A-LNCRNA轴与 抗性LSC来自我们的原则研究证明，表明i）动态和可逆的M6A 由脂肪质量和与肥胖相关蛋白（FTO）确定的甲基团有助于白血病细胞避免杀死TKI 导致TKI抗性； ii）有许多LNCRNA（注释）在抗性中差异表达 相对于敏感细胞。这些LNCRNA熊M6A图案中约有50％，并具有更改的M6A量 抗性细胞集体提出了特定于TKI耐药性的独特lncRNA特征，IS 由M6A甲基化调节； iii）在不反应的患者中上调这些与M6A相关的LNCRNA 对TKI的预后是较差的结果，而敲低的结果会损害耐药细胞的生长和呈现 对TKIS敏感的抗性细胞； iv）与敏感的细胞相比，高表达LSC标记 （CD117，CD44，CD25，CD133）其上调与M6A还原有关。我们的假设是 FTO-M6A-LNCRNA级联反应可能是控制LSC持久性TKI和新型吸毒的关键途径 目标根除持续的LSC，以提高TKI治疗速率。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耐药性中的lncRNA像差； 2）确定是否以及 动态M6a甲基团如何调节LSC持续到TKIS； 3）确定是否以及如何 FTO-M6A-LNCRNA CASCADE的药理靶向使用临床前白血病杀死持续的LSC 型号。拟议的研究在概念上是创新的，因为它针对了一种新途径（FTO-M6A-- lncrna级联）了解LSC持续到TKI和开发新方案以消除TKI 抗性LSC。拟议的研究很重要，因为发现将a）确定新途径（即 调节LSC持久性的FTO-M6A-LNCRNA级联反应，加深对LSC和LSC的理解 lncRNA功能； b）开发新方法（靶向FTO-M6A-LNCRNA级联）以消除持久性 LSC改善了难治性白血病患者的管理。