Development of companion diagnostics for dasatinib-based personalized therapy for T-ALL

开发基于达沙替尼的 T-ALL 个性化治疗伴随诊断

• 批准号: 10256123
• 负责人: Anton Iliuk
• 金额: $ 31万
• 依托单位: TYMORA ANALYTICAL OPERATIONS, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-02 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256123
• 关键词: ABL1 gene; Accounting; Acute Lymphocytic Leukemia; Acute T Cell Leukemia; Adult; Adult Acute Lymphocytic Leukemia; Adult Precursor T Lymphoblastic Leukemia; Algorithms; Antibodies; Biological Assay; Biological Markers; CRISPR screen; Cancer Etiology; Cancer cell line; Cell Culture Techniques; Cell Line; Cell Therapy; Cell model; Cessation of life; Child; Childhood; Clinical; Cytotoxic agent; Dasatinib; Dependence; Development; Diagnostic tests; Dose; Effectiveness; Exhibits; Feasibility Studies; Future; Hematologic Neoplasms; Human; Immunoassay; Immunotherapy; In Vitro; Inferior; Leukemic Cell; Lymphoid; Malignant Neoplasms; Measures; Methods; Mixed B- and T-Cell Leukemia; Modeling; Monitor; Mus; Network-based; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phosphoproteins; Phosphorylation; Population; Prognostic Marker; Proteins; Public Health; Quality of life; Quantitative Evaluations; Reaction; Recurrent disease; Regimen; Relapse; Resistance; Sampling; Sensitivity and Specificity; Signal Pathway; Signal Transduction; Small Business Innovation Research Grant; Survival Rate; T-Lymphocyte; Testing; Therapeutic; Toxic effect; Treatment Protocols; Treatment outcome; Tyrosine; Tyrosine Phosphorylation; United States National Institutes of Health; Validation; ZAP-70 Gene; activity marker; acute T-cell lymphoblastic leukemia cell; base; biomarker panel; chemotherapeutic agent; chemotherapy; companion diagnostics; cytotoxic; cytotoxicity; diagnostic panel; drug sensitivity; fusion gene; genome-wide; high risk; improved; in vivo; inhibitor/antagonist; interest; leukemia; leukemia treatment; molecular targeted therapies; mouse model; novel; novel therapeutics; patient derived xenograft model; patient population; personalized diagnostics; personalized medicine; phosphoproteomics; rapid technique; resistant strain; response; targeted agent; treatment response

PROJECT SUMMARY Acute lymphoblastic leukemia (ALL) is the most common malignancy in children and a leading cause of cancer-related death during childhood. ALL can arise in both lymphoid lineages, with B-ALL and T-ALL accounting for 85% and 15% of this cancer. T-ALL is associated with more aggressive presenting features and historically inferior treatment outcomes compared to B-ALL. Current T-ALL therapy largely relies on cytotoxic chemotherapeutic agents. In recent years, further intensification of chemotherapy has led to incremental increases in the cure rate of T-ALL, but is likely to have reached a plateau due to excessive toxicities (especially in the relapse setting). Moreover, in relapsed T-ALL, leukemia is markedly resistant to cytotoxic drugs. Unlike high-risk B-ALL for which cellular therapy such as CAR-T is highly effective, there are no immunotherapies available for T-ALL and patients with relapsed disease have a dismal five-year survival rate below 25%. Therefore, novel and molecularly targeted therapeutics are needed to improve both survival and quality of life for children with T-ALL. We have previously observed that in 64 T-ALL cases (43 children and 21 adults) profiled thus far 38% showed a striking sensitivity to dasatinib in vitro. In particular, the proportion of T-ALL sensitive to dasatinib is markedly higher in children than in adults (49% vs 14%, respectively). Dasatinib LC50 (drug concentration that kills 50% of leukemia cells) in these T-ALL cases were on par with that observed in BCR-ABL1 B-ALL. However, none of the dasatinib-sensitive T-ALL cases had ABL fusion nor did they respond to a more ABL-specific inhibitor. We demonstrated that LCK activation and phosphorylation level of its downstream targets are responsible for this sensitivity, and, more importantly, can be used to predict the effectiveness of dasatinib treatment in T-ALL cases. During this project, we will develop a companion diagnostic panel that can be used to predict sensitivity to dasatinib and ponatinib in a personalized manner. The following aims will be completed in the proposal: Aim #1. Comprehensive characterization of phosphorylation and activation state of LCK by LC-MS in T-ALL cells. Aim #2. Development of the PRM-MS and immunoassay-based methods for rapid quantitative analysis of p-LCK, p-CD247, and p-ZAP70. By the completion of this project, a companion diagnostic panel will be developed and validated with cell culture samples. This feasibility portion will enable a much more extensive validation of this personalized diagnostic test in PDX mice models and patient samples in Phase II.

项目概要 急性淋巴细胞白血病（ALL）是儿童最常见的恶性肿瘤，也是导致儿童罹患该病的主要原因 儿童时期与癌症相关的死亡。 ALL 可以出现在两种淋巴谱系中，其中 B-ALL 和 T-ALL占本癌的85%和15%。 T-ALL 与更具攻击性有关 与 B-ALL 相比，呈现出特征和历史上较差的治疗结果。当前T-ALL 治疗主要依赖细胞毒性化疗药物。近年来，进一步加大力度 化疗已导致 T-ALL 治愈率逐步提高，但可能会降低 T-ALL 的治愈率。 由于毒性过高（尤其是在复发情况下），达到了平台期。此外，在 复发性T-ALL、白血病对细胞毒药物有明显耐药性。与高风险 B-ALL 不同的是， CAR-T等细胞疗法非常有效，但目前尚无针对T-ALL的免疫疗法 疾病复发的患者五年生存率低至 25% 以下。因此，小说 需要分子靶向治疗来提高患者的生存率和生活质量 患有 T-ALL 的儿童。 我们之前观察到，迄今为止，在 64 例 T-ALL 病例（43 例儿童和 21 例成人）中 38% 的人在体外对达沙替尼表现出惊人的敏感性。特别是 T-ALL 敏感的比例 儿童对达沙替尼的使用率明显高于成人（分别为 49% 和 14%）。达沙替尼 LC50 这些 T-ALL 病例中的（杀死 50% 白血病细胞的药物浓度）与此相当 在 BCR-ABL1 B-ALL 中观察到。然而，对达沙替尼敏感的 T-ALL 病例均未出现 ABL 它们对 ABL 特异性抑制剂也没有反应。我们证明了 LCK 激活 其下游靶点的磷酸化水平是造成这种敏感性的原因，并且，更多 重要的是，可用于预测达沙替尼治疗 T-ALL 病例的有效性。期间 在这个项目中，我们将开发一个伴随诊断面板，可用于预测对 达沙替尼和普纳替尼以个性化方式进行。年内将完成以下目标 建议：目标#1。 LCK 磷酸化和激活状态的综合表征 T-ALL 细胞中的 LC-MS。目标#2。 PRM-MS 和基于免疫分析的方法的开发 p-LCK、p-CD247 和 p-ZAP70 的快速定量分析。该项目完成后， 将开发伴随诊断面板并用细胞培养样品进行验证。这个可行性 部分将使这种个性化诊断测试在 PDX 小鼠中得到更广泛的验证 第二阶段的模型和患者样本。