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.

项目概要 重大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 过去四十年中白血病（B-ALL）的发病率。在化疗中添加 ABL1 激酶抑制剂可显着 提高费城染色体阳性儿童和成人的无复发率和总体生存率 (Ph+) ALL 亚型。这一巨大的治疗成功归功于 Ph+ 白血病的致癌成瘾 BCR-ABL1 驱动的信号传导以及 ABL1 激酶抑制剂诱导深度细胞毒性的能力。 费城染色体样（Ph 样）ALL 是一种类似的高危白血病亚型，由各种非- BCR-ABL1 基因损伤同样会激活致癌激酶信号传导。 Ph 样 B-ALL 发生于 15-40% 患有 B-ALL 的儿童和成人，复发和死亡的风险特别高。我们确定了 CRLF2 重排 Ph 样中组成型激活的 JAK2/STAT5、PI3K 和 B 细胞受体样 (BCR) 信号传导 ALL，可以通过体外抑制 JAK1/2 (ruxolitinib)、PI3Kδ (idelalisib) 和 SRC (dasatinib) 来消除 以及临床前模型中的体内研究。然而，虽然三重抑制导致体外细胞完全死亡， 小鼠的白血病负担显着降低，我们怀疑这种治疗策略是否可以临床转化 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原因引起的漏洞 JAK2 激酶抑制并鉴定出 i) SGK1 的高活性，这是一种已知可诱导耐药性的分子 PI3Kα 突变乳腺癌患者中的 PI3K 抑制剂，ii) PTPN6 完全失活和下调，a 已知磷酸酶可下调 JAK2、PI3K 和 BCR 信号传导，以及 iii) DN​​A 损伤的积累 JAK2 抑制。我们假设 SGK1 的激活和 PTPN6 的失活是必需的机制 克服 JAK2 抑制剂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激活的 SGK1 可导致有效的体外细胞死亡。此外，我们 假设 JAK2 抑制剂诱导的 DNA 损伤是由 RAG 核酸内切酶复合物介导的， 它在 B 细胞分化过程中被激活，形成 BCR。研究表明，JAK2/STAT5 信号传导 下调分化 B 细胞以诱导增殖停滞，从而防止细胞死亡 V(D)J 重组过程中的细胞凋亡。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 的机制， 由于未修复的 DNA 损伤的积累，这可能会导致大量白血病细胞死亡。拟议的 研究旨在揭示以前未知的耐药机制并表征潜在的耐药性 Ph 样 B-ALL 细胞的生物学，最终建立临床前治疗方案，目标是预防 复发并治愈 Ph 样 B-ALL 患者。