Targeting SYK Tyrosine Kinase to Overcome Radiation Resistance in ALL

靶向 SYK 酪氨酸激酶以克服 ALL 的放射抗性

• 批准号: 8250355
• 负责人: FATIH M UCKUN
• 金额: $ 33.2万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8250355
• 关键词: Acute Lymphocytic Leukemia; Affect; Apoptosis; Apoptotic; B-Lymphocytes; BCL1 Oncogene; Binding; Biological Assay; Biological Markers; Biological Models; CD19 gene; Cells; Cessation of life; Chemosensitization; Childhood Leukemia; Development; Disease-Free Survival; Dose; Effectiveness; Electromagnetic Energy; Family; Foundations; Hematopoietic Stem Cell Transplantation; In Situ Nick-End Labeling; In Vitro; Inbred BALB C Mice; JAK3 gene; Janus kinase; Malignant Neoplasms; Modeling; Molecular Profiling; Molecular Target; Mus; Outcome; Pathway interactions; Patients; Phosphotransferases; Protein Tyrosine Kinase; Radiation; Radiation Tolerance; Radiation therapy; Radiation-Sensitizing Agents; Radiosensitization; Recurrence; Regimen; Relapse; Research; Research Project Grants; Resistance; SCID Mice; STAT3 gene; SYK gene; Safety; Signal Transduction; Site; Staining method; Stains; Testing; Therapeutic; Toxic effect; Translational Research; Treatment outcome; Whole-Body Irradiation; Work; Xenograft Model; annexin A5; chemotherapy; drug candidate; effective therapy; experience; improved; in vivo; inhibitor/antagonist; innovation; leukemia; member; novel; pre-clinical; protein expression; public health relevance; radiation resistance; repaired; success; therapy resistant; treatment program; treatment response

DESCRIPTION (provided by applicant): We are proposing to develop a new strategy to overcome radiation resistance in B- lineage acute lymphoblastic leukemia (ALL) using C-61, a novel SYK kinase substrate binding (P)-site inhibitor, for targeting and disrupting the anti-apoptotic SYK-STAT3 signaling network in leukemic B-cell precursors. It is our central working hypothesis that the treatment outcome of relapsed B-lineage ALL patients can be improved by inhibition of the SYK-STAT3 molecular target. This would be accomplished by using C-61 in combination with total body irradiation (TBI) in the context of HSCT. Under Specific Aim 1, we will examine the effects of the SYK P-site inhibitor C-61 on in vitro radiation resistance of primary ALL cells from relapsed B-lineage ALL patients using quantitative flow cytometric apoptosis assays and clonogenic assays. We hypothesize that C- 61 will markedly enhance radiation-induced apoptosis of primary B-lineage ALL cells and augment radiation- induced death of their clonogenic fraction by increasing their radiation sensitivity and impairing their capacity to repair sublethal radiation damage. Under Specific Aim 2, we will examine the effects of C-61 on in vivo radiation resistance of primary ALL cells from relapsed B-lineage ALL patients using SCID mouse xenograft models of relapsed B-lineage ALL and sublethal total body irradiation (TBI). Our hypothesis is that C-61 plus TBI regimens will be more effective than TBI alone in improving the event-free survival outcome of SCID mice challenged with primary B-lineage ALL cells. Under Specific Aim 3, we will examine the association between the kinase expression profiles of primary ALL cells from relapsed B-lineage ALL patients and their in vitro as well as in vivo radiation resistance. In an effort aimed at identifying a composite biomarker profile that will help select patients most likely to benefit from C-61, we will correlate the kinase protein expression and activity levels of SYK, BTK, and JAK kinases of primary B-lineage ALL cells with their radiation resistance, sensitivity to C-61 induced radiosensitization in vitro, as well as C-61 induced potentiation of the anti-leukemic potency of sublethal TBI in vivo. Under Specific Aim 4, we will study the efficacy and safety of C-61 containing single dose TBI regimens at both sublethal (2 Gy) as well as clinically applied (7 Gy) total radiation dose levels in a syngeneic murine HSCT model of radiation-resistant BCL-1 murine B-lineage leukemia. We will evaluate the efficacy and safety of TBI at doses ranging from 2-10 Gy in combination with C-61 in BALB/c mice inoculated with 1x106 BCL-1 cells in the context of syngeneic BMT. We hypothesize that the addition of C-61 will not increase the non-hematologic toxicity of TBI, while markedly potentiating its anti-leukemic efficacy. We anticipate that the incorporation of C-61 into the pre-HSCT TBI regimens of patients with relapsed B-lineage ALL will improve their treatment response and survival outcome. The proposed research has the potential provide the foundation for the development of paradigm-shifting HSCT strategies that employ C-61 containing novel TBI regimens. PUBLIC HEALTH RELEVANCE: Currently, the major challenge in the treatment of childhood leukemia is to cure patients who experience a recurrence of their cancer despite intensive chemotherapy. The purpose of the proposed research is the development of an effective treatment program against chemotherapy- and radiation therapy-resistant childhood leukemia employing a new anti-leukemia drug candidate. The successful completion of this research project may provide the foundation for therapeutic innovation against childhood leukemia.

描述（由申请人提供）：我们建议使用一种新的策略来克服B-谱系急性淋巴细胞性白血病（ALL），使用C-61（一种新型的Syk激酶底物结合（P） - 位置抑制剂，用于靶向和破坏抗popoptotic Syksy Syksy Sykstotic Syk-Stat3 Signal 3网络中的抑制剂。我们的核心工作假设是，复发B-linege的治疗结果可以通过抑制SYK-STAT3分子靶标的改善所有患者。这将通过在HSCT的背景下将C-61与总体辐照（TBI）结合使用来实现。在特定的目标1下，我们将使用定量的水流细胞量尺寸凋亡测定法和克隆发生测定法检查SYK P位抑制剂C-61对所有细胞的体外辐射耐药性的影响。我们假设C-61将显着增强辐射诱导的原代B-linege所有细胞的凋亡，并通过增加其辐射敏感性并损害修复层次辐射损伤的能力，从而增强其克隆性分数的辐射诱导死亡。在特定的目标2下，我们将使用复发的B-Linege的原代细胞中C-61对所有患者的体内辐射耐药性的影响，使用Relapsed B-Linege的SCID小鼠异种移植模型全部和余生的全身辐照（TBI）。我们的假设是，C-61 Plus TBI方案将比单独使用TBI更有效，以改善针对所有细胞挑战的SCID小鼠的无事件生存结果。在特定的目标3下，我们将检查所有患者复发的所有患者及其体外和体内耐药性的基本细胞的激酶表达谱之间的关联。为了确定将有助于选择最有可能从C-61中受益的患者的复合生物标志物概况的努力，我们将将SYK，BTK和JAK激酶的激酶蛋白表达和活性水平相关联，将所有细胞的所有细胞的jak激酶与辐射的耐药性与C-61诱导的辐射性抗性相关联，并在C-61中诱导的potitiation和C-61的potitiation和C-61的potitiation and c-61 potertiation and and c-61 poteciation and and c-61 poteciation and and c-61 poteciation以及C-61的均匀性。子体内tbi。在特定目标4下，我们将研究含有单剂量TBI治疗方案的C-61（2 Gy）以及临床应用（7 Gy）总辐射剂量水平在辐射耐药的Bcl-1鼠BCL-1鼠B-Lineage Leukemia中的临床应用（7 Gy）总辐射剂量水平。我们将评估TBI在剂量下的疗效和安全性，范围从2-10 Gy与C-61结合使用BALB/C小鼠中的C-61，在合成BMT的背景下接种了1x106 Bcl-1细胞。我们假设添加C-61不会增加TBI的非血液毒性，同时显着增强其抗白血病功效。我们预计将C-61掺入复发B-Linege患者的HSCT前TBI方案中，所有这些都将改善其治疗反应和生存结果。拟议的研究具有潜力为开发范式转移HSCT策略的基础，这些策略采用了含有新型TBI方案的C-61。 公共卫生相关性：目前，治疗儿童白血病的主要挑战是治疗尽管化学疗法进行了严重化学疗法，但经历了癌症的患者。拟议研究的目的是开发一种有效的治疗计划，以采用新的抗白血病药物候选药物，以抗化疗和放射治疗的儿童白血病。该研究项目的成功完成可能为针对儿童白血病的治疗创新提供基础。