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; 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.

项目摘要 急性淋巴细胞白血病（ALL）是儿童中最常见的恶性肿瘤，也是主要原因 童年时期与癌症有关的死亡。在两个淋巴谱系中都可以出现所有，b-all和 T-All占该癌症的85％和15％。 T-All与更具侵略性相关 与B-All相比，呈现特征和历史较低的治疗结果。当前T-ALL 治疗主要依赖于细胞毒性化学治疗剂。近年来，进一步加强 化学疗法导致T-all的治愈率增加，但可能具有 由于过度毒性（尤其是在复发环境中），达到了高原。而且，在 复发的T-ALL，白血病明显抗细胞毒性药物。与高风险B-all不同 诸如CAR-T之类的细胞疗法非常有效，没有免疫疗法可用于T-All 复发性疾病的患者的五年生存率低于25％。因此，新颖 需要分子靶向治疗，以改善生存和生活质量 有T-All的孩子。 我们以前已经观察到，到目前为止，有64例T-ALL病例（43名儿童和21名成年人）构图 38％的体外表现出对达沙替尼的敏感性。特别是T-ALL敏感的比例 儿童的达沙替尼显着高于成年人（分别为49％和14％）。达沙替尼LC50 （杀死50％白血病细胞的药物浓度）在这些T-ALL情况下与此相提并论 在BCR-ABL1 B-ALL中观察到。但是，dasatinib敏感的T-All Case均未有ABL 融合也没有对更ABL特异性抑制剂的反应。我们证明了LCK激活 其下游靶标的磷酸化水平是该灵敏度的原因，并且更多 重要的是，可以用来预测达沙替尼治疗在T-All情况下的有效性。期间 这个项目，我们将开发一个伴侣诊断面板，可用于预测对 达沙替尼和庞蒂尼以个性化的方式。以下目标将在 提案：目标＃1。 LCK对LCK的磷酸化和激活状态的全面表征 T-ALL细胞中的LC-MS。目标＃2。开发PRM-MS和基于免疫测定方法的方法 P-LCK，P-CD247和P-ZAP70的快速定量分析。根据该项目的完成， 伴侣诊断面板将通过细胞培养样品开发和验证。这种可行性 部分将对PDX小鼠进行该个性化诊断测试的更广泛验证 II期模型和患者样品。