Targeting Kinase Inhibitor Induced Signaling Plasticity in Patients with Ph-Like ALL

针对 Ph 样 ALL 患者中激酶抑制剂诱导的信号可塑性

• 批准号: 10584455
• 负责人: Christian Hurtz
• 金额: $ 19.31万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-04 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10584455
• 关键词: ABL1 gene; Acute Lymphocytic Leukemia; Adult; Age; Animals; Apoptosis; B cell differentiation; B-Cell Activation; B-Cell Acute Lymphoblastic Leukemia; B-Cell Antigen Receptor; B-cell precursor acute lymphoblastic leukemia cell; Bioinformatics; Biology; Breast Cancer Patient; CHEK1 gene; Cell Death; Cell Differentiation Induction; Cell Line; Cell Proliferation; Cell Survival; Cells; Cellular Stress; Cessation of life; Child; Complex; Critical Pathways; Cytokine Receptors; DNA; DNA Damage; DNA Double Strand Break; Dasatinib; Data; Development; Down-Regulation; Drug resistance; Enzyme Induction; Enzymes; Gene Expression; Gene Expression Profile; Genetic; Genetic Heterogeneity; Goals; Heterochromatin; Human; In Vitro; JAK1 gene; JAK2 gene; Lesion; Leukemic Cell; Mediating; Molecular; Mus; Mutate; Mutation; Oncogenes; Oncogenic; PI3K/AKT; PIK3CG gene; PTPN6 gene; Patient-Focused Outcomes; Patients; Ph+ ALL; Philadelphia Chromosome; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Pre-Clinical Model; Proliferating; Proto-Oncogene Proteins c-akt; Receptor Signaling; Relapse; Research; Sampling; Signal Induction; Signal Pathway; Signal Transduction; Signaling Molecule; Source; Stat5 protein; Techniques; Testing; Therapeutic; Toxic effect; Training; Treatment Protocols; Uncertainty; V(D)J Recombination; acute lymphoblastic leukemia cell; addiction; cell type; chemotherapy; clinical translation; cytokine; cytotoxicity; defined contribution; endonuclease; experimental study; flexibility; high risk; human disease; improved; improved outcome; in vivo; inhibitor; innovation; kinase inhibitor; leukemia; leukemia treatment; loss of function; mortality; mouse model; novel therapeutic intervention; patient derived xenograft model; pharmacologic; pre-clinical; preclinical study; prevent; protective pathway; relapse prevention; relapse risk; repaired; response; src-Family Kinases; success; synergism; targeted treatment; therapeutic target; therapeutically effective; translational study; treatment strategy

Project Summary Major scientific and therapeutic advances have improved outcomes of patients with B-cell acute lymphoblastic leukemia (B-ALL) over the last four decades. Addition of ABL1 kinase inhibitors to chemotherapy has significantly improved relapse-free and overall survival of children and adults with the Philadelphia chromosome positive (Ph+) ALL subtype. This great treatment success has been attributed to oncogenic addiction of Ph+ leukemias to BCR-ABL1-driven signaling and the ability of ABL1 kinase inhibitors to induce profound cytotoxicity. Philadelphia chromosome-like (Ph-like) ALL is an analogous high-risk leukemia subtype driven by various non- BCR-ABL1 genetic lesions that similarly activate oncogenic kinase signaling. Ph-like B-ALL occurs in 15-40% of children and adults with B-ALL and is associated with a particularly high risk of relapse and death. We identified constitutively activated JAK2/STAT5, PI3K and B cell receptor-like (BCR) signaling in CRLF2-rearranged Ph-like ALL, which can be abrogated with JAK1/2 (ruxolitinib), PI3Kδ (idelalisib), and SRC (dasatinib) inhibition in vitro and in vivo in preclinical models. However, while the triple inhibition results in complete cell death in vitro and significant decrease of the leukemia burden of mice, we doubt that this treatment strategy is clinically translatable for patients given potential for appreciable toxicity. We thus focused on discovering vulnerabilities induced by JAK2-kinase inhibition and identified i) high activity of SGK1, a molecule known to induce drug resistance to PI3K inhibitors in PI3Kα-mutated breast cancer patients, ii) complete inactivity and downregulation of PTPN6, a phosphatase known to downregulate JAK2, PI3K, and BCR signaling, and iii) accumulation of DNA damage with JAK2 inhibition. We hypothesized that activation of SGK1 and inactivation of PTPN6 are required mechanisms to overcome JAK2 inhibitor-induced signaling inhibition. Strikingly, our preliminary data demonstrated that targeting activated SGK1 in JAK2 inhibitor treated samples results in potent in vitro cell death. Furthermore, we hypothesized that the JAK2-inhibitor induced DNA-damage is mediated by the RAG endonuclease complex, which is activated during B cell differentiation to form the BCR. It has been shown that JAK2/STAT5 signaling is downregulated in differentiating B cells to induce a proliferation arrest, which prevents cells from dying from apoptosis during V(D)J recombination. Our preliminary data demonstrate that we can exploit this conserved mechanism by targeting the DNA-damage response molecule CHK1 in JAK2 inhibitor treated Ph-like ALL cells, which may result in massive leukemic cell death due to accumulation of unrepaired DNA damage. The proposed study aims to uncover previously unknown mechanisms of drug resistance and to characterize the underlying biology of Ph-like B-ALL cells to ultimately establish preclinical treatment protocols with the goal to prevent relapse and to cure patients with Ph-like B-ALL.

项目摘要 主要的科学和治疗进展改善了B细胞急性淋巴细胞白血病患者的预后 白血病（B-ALL）在过去的40年。在化疗中加入ABL1激酶抑制剂， 改善费城染色体阳性儿童和成人的无复发生存率和总生存率 (Ph+）ALL亚型。这种巨大的治疗成功归因于Ph+白血病的致癌成瘾 BCR-ABL1驱动的信号传导和ABL1激酶抑制剂诱导显著细胞毒性的能力。 费城染色体样（Ph样）ALL是一种类似的高风险白血病亚型，由各种非 BCR-ABL1基因病变同样激活致癌激酶信号。Ph样B-ALL发生在15 - 40%的 B-ALL的儿童和成人与复发和死亡的风险特别高相关。我们确定 CRLF 2重排的Ph样细胞中组成性激活JAK2/STAT 5、PI3 K和B细胞受体样（BCR）信号传导 ALL，可通过JAK 1/2（ruxolitinib）、PI3K δ（idelalisib）和SRC（达沙替尼）体外抑制作用消除 以及在临床前模型中的体内。然而，虽然三重抑制导致体外完全细胞死亡， 小鼠白血病负荷的显著降低，我们怀疑这种治疗策略在临床上是可转化的 对于可能出现明显毒性的患者。因此，我们专注于发现由以下因素引起的漏洞： i）SGK1的高活性，SGK1是一种已知诱导对JAK2-激酶的耐药性的分子， PI3K α突变乳腺癌患者中的PI3K抑制剂，ii）PTPN 6完全失活和下调，a 已知下调JAK2、PI3K和BCR信号传导的磷酸酶，和iii）DNA损伤的积累， JAK2抑制。我们假设SGK1的激活和PTPN6的失活是需要的机制 以克服JAK2激酶诱导的信号传导抑制。引人注目的是，我们的初步数据表明， 在JAK2抑制剂处理的样品中靶向活化的SGK1导致有效的体外细胞死亡。而且我们 假设JAK 2抑制剂诱导的DNA损伤是由RAG内切核酸酶复合物介导的， 其在B细胞分化过程中被激活以形成BCR。已经表明JAK2/STAT5信号传导是 在分化中的B细胞中下调以诱导增殖停滞，这防止细胞死于 在V（D）J重组期间的细胞凋亡。我们的初步数据表明，我们可以利用这种保守的 通过靶向JAK2抑制剂处理的Ph样ALL细胞中的DNA损伤反应分子CHK1， 这可能由于未修复的DNA损伤的积累而导致大量白血病细胞死亡。拟议 这项研究旨在揭示以前未知的耐药机制，并描述潜在的耐药机制。 Ph样B-ALL细胞的生物学，以最终建立临床前治疗方案， 复发和治愈Ph样B-ALL患者。