Lapatinib and Obatoclax combination therapy

拉帕替尼和 Obatoclax 联合治疗

• 批准号: 8600243
• 负责人: PAUL DENT
• 金额: $ 32.84万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-07 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8600243
• 关键词: Animals; Apoptotic; Autophagocytosis; BCL1 Oncogene; BCL2 gene; BH3 Domain; Breast Cancer Cell; Breast Carcinoma; Cancer Patient; Caspase; Catalytic Domain; Cells; Cessation of life; Colon Carcinoma; Combined Modality Therapy; Cytosol; Data; Diagnosis; Dose; Drops; Drug Combinations; EGFR gene; ERBB2 gene; Endoplasmic Reticulum; Estrogen Receptors; Evaluation; FDA approved; Family; Family member; Fatty acid glycerol esters; Growth; Growth Factor Receptors; Health Benefit; Human Mammary Carcinoma; Image; Immune; In Vitro; Inbred BALB C Mice; Ionizing radiation; Laboratory Study; Malignant Epithelial Cell; Mammary Neoplasms; Mammary gland; Mediating; Mitochondria; Molecular; Mouse Mammary Tumor Virus; Mus; Mutation; Neoplasm Metastasis; Nude Mice; Outcome; PMAIP1 gene; Patients; Pharmaceutical Preparations; Phase; Phase II Clinical Trials; Phosphotransferases; Protein Family; Public Health; Pump; Radiation; Radiation Tolerance; Radiation-Sensitizing Agents; Recurrence; Resistance; Resistance development; Rodent; Small Interfering RNA; Survival Rate; System; Testing; Toxic effect; Translations; Tumor Cell Line; Vesicle; apoptosis inducing factor; base; calreticulin; cancer cell; cancer therapy; cell killing; cell type; cytochrome c; improved; in vivo; inhibition of autophagy; inhibitor/antagonist; killings; knock-down; lapatinib; malignant breast neoplasm; mitochondrial autophagy; mutant; neoplastic cell; public health relevance; tumor; tumor growth

DESCRIPTION (provided by applicant): Prior studies from this laboratory have demonstrated that resistance to the lethal effects of the FDA approved ERBB1 / ERBB2 inhibitor Lapatinib in carcinoma cells is mediated by increased expression of pro-survival BCL-2 family proteins e.g. MCL-1 and BCL-XL. More recently we have gone on to demonstrate that inhibition of MCL-1 and BCL-XL using molecular approaches or using the BCL-2 family inhibitor Obatoclax (GX15-070), a drug in phase II trials, promotes Lapatinib toxicity and reverts Lapatinib resistance. These studies were performed in vitro using a wide range breast and colon cancer cells; drug combination toxicity did not simplistically correlate with ERBB1/2 over-expression. Toxicity, also, was not suppressed by mutant active K- RAS expression or loss of p53 function in the colon cancer cells. This application has three hypotheses: We hypothesize that Lapatinib + Obatoclax treatment causes a toxic form of mitochondrial autophagy. We hypothesize that Lapatinib + Obatoclax treatment interact in vivo in a greater than additive fashion to promote mammary tumor cell killing; reducing tumor growth and metastatic spread. We hypothesize that (Lapatinib + Obatoclax) toxicity in vivo will be magnified in a greater than additive fashion by exposure of the tumor to low doses of ionizing radiation. Specific Aim 1. Determine the molecular mechanisms by which Obatoclax and Lapatinib interact to cause toxic autophagy. The molecular mechanisms of mitochondrial mediated autophagy are not fully understood. We will determine whether Obatoclax causes rapid autophagic degradation of mitochondria via BAK and NOXA (and their altered association with BCL-2 family proteins), with a rapid drop in cellular ATP levels, rather than degradation of the endoplasmic reticulum. We will assess the association of ATG8 and ATG6 (Beclin1) with mitochondrial e.g. HSP60, and ER markers e.g. calreticulin. Specific Aim 2. Determine using orthotopic systems in athymic and immune competent mice whether Obatoclax and Lapatinib interact to kill tumor cells, suppress tumor growth and prolong animal survival. We propose to use multiple tumor cell types to definitively test whether Lapatinib and Obatoclax interact in vivo to reduce tumor growth. We will use human mammary carcinoma BT474 cells growing in the mammary fat pad of athymic mice. We will grow rodent mammary carcinoma MMTV-HER2 cells and 4T1in the mammary fat pad of immune competent syngeneic FVB/NJ and BALB/c mice, respectively; for 4T1 cells we will determine the impact of drug treatment on metastatic tumor cell spread and growth via bioluminescent imaging. Specific Aim 3. Determine whether (Lapatinib + Obatoclax) therapy acts as a radiosensitizer in vivo. We will determine whether (Lapatinib + Obatoclax) treatment enhances tumor cell radiosensitivity in vitro and in vivo. Based on tumor growth / viability data obtained in Aim 2, one of the tumor cell lines will be selected for in vivo evaluation with (Lapatinib + Obatoclax) treatment and radiation exposure in aim 3. Lapatinib is an FDA approved ERBB1 / ERBB2 inhibitor. Obatoclax is in phase II trials. The successful completion of the studies proposed in the application would rapidly facilitate phase I translation of this combination therapy in breast cancer patients.

描述（由申请人提供）：该实验室的先前研究表明，FDA认可的ERBB1 / ERBB2抑制剂Lapatinib在癌细胞中对致命作用的抗性是通过增强促生抗菌Bcl-2家族蛋白的表达来介导的。 MCL-1和BCL-XL。最近，我们继续证明，使用分子方法或使用Bcl-2家族抑制剂obatoclax（GX15-070）对MCL-1和BCL-XL的抑制作用，这是II期试验中的一种药物，促进Lapatinib毒性并恢复Lapatinib的耐药性。这些研究是在体外使用广泛的乳腺癌和结肠癌细胞进行的。药物组合毒性与ERBB1/2的过表达无关。同样，毒性也不会被突变活性K-ras表达或在结肠癌细胞中p53功能的丧失所抑制。该应用具有三个假设：我们假设拉帕替尼 + obatoclax处理会引起线粒体自噬的有毒形式。我们假设Lapatinib + Obatoclax治疗以比添加剂方式更大的体内相互作用，以促进乳腺肿瘤细胞杀死。减少肿瘤生长和转移扩散。我们假设（Lapatinib + obatoclax）在体内的毒性将通过暴露于低剂量的电离辐射，以大于添加剂的方式放大。具体目的1。确定obatoclax和lapatinib相互作用以引起有毒自噬的分子机制。线粒体介导的自噬的分子机制尚未完全了解。我们将确定obatoclax是否会通过BAK和NOXA（及其与Bcl-2家族蛋白的改变改变）导致线粒体的快速自噬降解，并导致细胞ATP水平迅速下降，而不是内质网的降解。我们将评估ATG8和ATG6（Beclin1）与线粒体的关联，例如HSP60和ER标记，例如钙网蛋白。具体目的2。确定在无胃和免疫能力的小鼠中使用原位系统，无论obatoclax和lapatinib相互作用以杀死肿瘤细胞，抑制肿瘤的生长和延长动物的存活。我们建议使用多种肿瘤细胞类型来确定测试Lapatinib和Obatoclax是否在体内相互作用以减少肿瘤生长。我们将使用在无胸腺小鼠的乳腺脂肪垫中生长的人类乳腺癌BT474细胞。我们将分别生长啮齿动物乳腺癌MMTV-HER2细胞和4T1中的4T1在免疫胜任的合成性FVB/NJ和BALB/C小鼠的乳腺脂肪垫中；对于4T1细胞，我们将通过生物发光成像确定药物治疗对转移性肿瘤细胞扩散和生长的影响。具体目的3。确定（Lapatinib + obatoclax）治疗是否在体内充当放射敏化剂。我们将确定（Lapatinib + obatoclax）治疗是否会增强体外和体内肿瘤细胞放射敏感性。基于AIM 2中获得的肿瘤生长 /生存能力数据，将选择一种肿瘤细胞系进行体内评估（Lapatinib + obatoclax）治疗和AIM 3中的辐射暴露。Lapatinib是FDA批准的ERBB1 / ERBB2抑制剂。 Obatoclax正在II期试验中。应用程序中提出的研究的成功完成将迅速促进乳腺癌患者这种联合疗法的I期翻译。