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）：我们提议开发一种新策略，使用 C-61（一种新型 SYK 激酶底物结合 (P) 位点抑制剂）来克服 B 系急性淋巴细胞白血病 (ALL) 的辐射抗性，用于靶向和破坏白血病 B 细胞前体中的抗凋亡 SYK-STAT3 信号网络。我们的核心工作假设是，通过抑制 SYK-STAT3 分子靶标可以改善复发 B 系 ALL 患者的治疗结果。这可以通过在 HSCT 背景下使用 C-61 与全身照射 (TBI) 相结合来实现。在具体目标 1 下，我们将使用定量流式细胞术凋亡测定和克隆形成测定来检查 SYK P 位点抑制剂 C-61 对来自复发 B 系 ALL 患者的原代 ALL 细胞的体外辐射耐受性的影响。我们假设C-61将显着增强辐射诱导的原代B系ALL细胞的细胞凋亡，并通过增加其辐射敏感性和削弱其修复亚致死辐射损伤的能力来增强辐射诱导的其克隆形成部分的死亡。在具体目标 2 下，我们将使用复发 B 系 ALL 的 SCID 小鼠异种移植模型和亚致死全身照射 (TBI) 来检查 C-61 对复发 B 系 ALL 患者的原代 ALL 细胞体内辐射耐受性的影响。我们的假设是，C-61 加 TBI 方案比单独 TBI 更有效地改善接受原代 B 系 ALL 细胞攻击的 SCID 小鼠的无事件生存结果。在具体目标 3 下，我们将检查复发 B 系 ALL 患者的原代 ALL 细胞的激酶表达谱与其体外和体内辐射耐受性之间的关联。为了确定一个复合生物标志物谱，帮助选择最有可能从 C-61 中受益的患者，我们将原代 B 系 ALL 细胞的 SYK、BTK 和 JAK 激酶的激酶蛋白表达和活性水平与它们的辐射耐受性、对 C-61 诱导的体外放射增敏的敏感性以及 C-61 诱导的抗白血病效力的增强联系起来。 体内亚致死 TBI。在具体目标 4 下，我们将在抗辐射 BCL-1 鼠 B 系白血病的同基因鼠 HSCT 模型中研究含单剂量 TBI 方案的 C-61 在亚致死 (2 Gy) 和临床应用 (7 Gy) 总辐射剂量水平下的有效性和安全性。我们将在同基因 BMT 的情况下，在接种 1x106 BCL-1 细胞的 BALB/c 小鼠中评估 2-10 Gy 剂量范围的 TBI 与 C-61 的疗效和安全性。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 纳入复发 B 系 ALL 患者的 HSCT 前 TBI 方案中将改善他们的治疗反应和生存结果。拟议的研究有可能为开发范式转变的 HSCT 策略奠定基础，该策略采用包含新型 TBI 方案的 C-61。 公共卫生相关性：目前，儿童白血病治疗的主要挑战是治愈尽管进行强化化疗但癌症复发的患者。拟议研究的目的是利用一种新的抗白血病候选药物开发一种有效的治疗方案来对抗化疗和放射治疗耐药的儿童白血病。该研究项目的成功完成可能为儿童白血病的治疗创新奠定基础。