Bmi-1, a potential therapeutic target in ovarian cancer

Bmi-1，卵巢癌的潜在治疗靶点

基本信息

批准号：
9067819
负责人：
Resham Bhattacharya
金额：
$ 31.15万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-07 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9067819
关键词：
Acetylcysteine Address Apoptosis B-Cell Lymphomas CASP3 gene Cancer Patient Carboplatin Cell Line Cells Cisplatin Clinical DNA Damage Data Development Disease Down-Regulation Doxycycline Drug resistance Epithelial Generations Genes Goals Growth Gynecologic HRAS gene Homeostasis Immunohistochemistry In Vitro Lymphocyte Malignant Neoplasms Malignant neoplasm of ovary Mediating Messenger RNA Metastatic breast cancer Methods MicroRNAs Mitochondria Modeling Molecular Molecular Target Moloney Leukemia Virus Morbidity - disease rate Mus Neoplastic Cell Transformation Nodule Oncogenic Operative Surgical Procedures Ovarian Oxidation-Reduction Oxygen Oxygen Consumption Paclitaxel Pathway interactions Patient-Focused Outcomes Patients Phosphorylation Play Polycomb Process Production Reactive Oxygen Species Recurrent Malignant Neoplasm Relapse Reporting Resistance Reverse Transcriptase Polymerase Chain Reaction Role Sampling Small Interfering RNA Staging Stem cells Testing Therapeutic Tissues Treatment Efficacy Tumor Debulking United States National Institutes of Health Western Blotting cancer cell carcinogenesis chemotherapy combat drug sensitivity gene product in vivo innovation intraperitoneal knock-down mortality mouse model nanoliposome neoplastic cell novel therapeutic intervention ovarian neoplasm overexpression pre-clinical response self-renewal small hairpin RNA stem therapeutic target tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Undoubtedly ovarian cancer is a vexing, incurable disease for patients with recurrent cancer and has been declared one of four untreatable cancers by NIH. Effective therapeutic options are quite limited; and, as a result, morbidity worsens and survival shortens dramatically. In view of this unfortunate dilemma, our goal in this proposal is to explore the therapeutic potential of Bmi-1 in ovarian carcinogenesis. In this context, determining the mechanism by which loss of Bmi-1 sensitizes the cancer cells to cisplatin would be important for development of new therapeutic strategies to combat ovarian cancer. Bmi-1 (B lymphoma mouse Moloney leukemia virus insertion region) regulates the self-renewal of normal stem and progenitor cells. In addition, overexpression of Bmi-1 and its correlation with clinical grade/stage has been reported in a variety of cancers including ovarian cancer. We demonstrated that, downregulation of Bmi-1 decreased clonogenicity, proliferation and sensitized ovarian cancer cells to cisplatin-induced apoptosis. Bmi- 1 causes neoplastic transformation of lymphocytes and co-operates with H-Ras giving rise to metastatic breast cancer in mice, all strongly suggesting an oncogenic role in epithelial malignancies. Hence the development of new therapeutic strategies to inactivate Bmi-1 to combat ovarian cancer most likely will have a strong impact on clinical outcome and patient management. Here we postulate that Bmi-1 plays an important role in ovarian cancer growth and can be manipulated to increase sensitivity to cisplatin. Furthermore, we will investigate the molecular mechanism by which downregulation of Bmi-1 sensitizes the ovarian cancer cells to cisplatin to develop new therapeutic strategies. To address our hypothesis, we propose the following aims: Aim1: To determine the mechanism by which loss of Bmi-1 enhances drug-sensitivity. Rationale and hypothesis: We recently demonstrated that downregulation of Bmi-1 by siRNA or miR significantly enhanced cisplatin-induced apoptosis in ovarian cancer cells including the cisplatin resistant CP- 70. Importantly use of the oxygen scavenger N-Acetylcysteine (NAC) abolished cisplatin induced apoptosis suggesting involvement of reactive oxygen species (ROS) and/or NFkappaB (NF:B) in this process. Furthermore, cisplatin treatment in Bmi-1 knockdown cells led to significant engagement of the DNA damage response (DDR) pathway leading to apoptosis. Hence, we hypothesize that silencing Bmi-1 enhances cisplatin-induced apoptosis through NF:B and/or ROS mediated activation of the DDR pathway. Approach: Therefore (i) NF:B activation status, (ii) total and mitochondrial ROS generation in Bmi-1 silenced cisplatin treated cells wil be determined. Mitochondrial parameters such as oxygen consumption, ATP production, gene products involved in redox homeostasis will be studied. (iii) DDR engagement leading to apoptosis will be studied by changes in phosphorylation or localization of different markers including ATM1, ATR, Chk1 and Chk2. Cleavage of Caspase-3, 8, 9 and PARP will also be determined. Aim2: To determine the therapeutic potential of Bmi-1 knockdown as a modulator of chemosensitivity to cisplatin in ovarian cancer in vivo. Rationale and hypothesis: We recently demonstrated that ovarian tumor growth and nodule formation were significantly reduced upon delivery of Bmi-1 siRNA by nanoliposome in vivo in an A-2780/CP-20 intraperitoneal model. Downregulation of Bmi-1 by siRNA or microRNA-15a/16 that directly target Bmi-1 mRNA resulted in inhibition of proliferation, clonogenicity and sensitized cells to cisplatin-induced apoptosis. Hence, we hypothesize that downregulation of Bmi-1 will inhibit ovarian tumor growth and sensitize the tumor cells to cisplatin in a preclinical mouse model. Approach: We will test the therapeutic efficacy in orthotopic intraperitoneal ovarian tumor mouse models generated by injecting A-2780 or CP-70 (cisplatin resistant) cels. To simulate treatment of advanced smal- volume disease, cisplatin and or nanoliposomal conjugates of siRNA/miR will be administered intraperitoneally 1 week after tumor inoculation and continued for 4 weeks. We will determine (i) if knockdown of Bmi-1 by siRNA, (ii) if knockdown of Bmi-1 by miR-15a/16 inhibits tumor growth and sensitizes cells to cisplatin in vivo; (iii) Alternatively, doxycycline inducible conditional knockdown of Bmi-1 by shRNA in A-2780 and CP-70 cells will be utilized; (iv) Validate the molecular targets of the DDR and apoptosis pathway as in Aim 1, in tumor cells in vivo by Western blot analysis, RT-PCR and immunohistochemistry (IHC). The proposed studies will validate Bmi-1 as an important new target using readily translatable nanoliposomal delivery methods with potential for therapy even in chemoresistant ovarian cancer.

描述（由申请人提供）：毫无疑问，卵巢癌对复发性癌症患者来说是一种令人烦恼的，无法治愈的疾病，已被NIH宣布为四种不可治疗的癌症之一。有效的治疗选择非常有限；因此，发病率恶化和生存会大大缩短。鉴于这种不幸的困境，我们在该提案中的目标是探索BMI-1在卵巢癌发生中的治疗潜力。在这种情况下，确定BMI-1损失使癌细胞对顺铂敏感的机制对于制定对抗卵巢癌的新治疗策略至关重要。 BMI-1（bmi-1小鼠小鼠白血病病毒插入区域）调节正常茎和祖细胞的自我更新。此外，在包括卵巢癌在内的各种癌症中，BMI-1的过表达及其与临床等级/阶段的相关性。我们证明，BMI-1的下调降低了克隆性，增殖和敏化卵巢癌细胞对顺铂诱导的细胞凋亡。 BMI-1引起淋巴细胞的肿瘤转化，并与H-RAS合作，导致小鼠转移性乳腺癌，这都强烈表明在上皮恶性肿瘤中具有致癌作用。因此，开发新的治疗策略使BMI-1灭活卵巢癌很可能会对临床结果和患者管理产生强大的影响。在这里，我们假设BMI-1在卵巢癌的生长中起着重要作用，并且可以操纵以提高对顺铂的敏感性。此外，我们将研究BMI-1下调的分子机制，使卵巢癌细胞对顺铂敏感以开发新的治疗策略。为了解决我们的假设，我们提出以下目的：AIM1：确定BMI-1损失增强药物敏感性的机制。理由和假设：我们最近证明，siRNA或MIR对BMI-1的下调显着增强了顺铂诱导的卵巢癌细胞中凋亡的凋亡，包括抗顺铂的CP-70。和/或NFKAPPAB（NF：B）在此过程中。此外，在BMI-1敲除细胞中的顺铂治疗导致DNA损伤反应（DDR）途径的大量参与导致凋亡。因此，我们假设沉默BMI-1通过NF：B和/或ROS介导的DDR途径激活增强了顺铂诱导的凋亡。方法：（i）NF：B激活状态，（ii）BMI-1沉默的顺铂处理细胞中的总和线粒体ROS产生将确定。将研究线粒体参数，例如氧气消耗，ATP产生，涉及氧化还原稳态的基因产品。（iii）通过磷酸化或包括ATM1，ATR，CHK1和CHK2的不同标记的磷酸化或定位的变化来研究导致凋亡的DDR参与度。 caspase-3、8、9和PARP的切割也将确定。 AIM2：确定BMI-1敲低作为体内卵巢癌化学敏感性的调节剂的治疗潜力。基本原理和假设：我们最近证明，纳米脂体在A-2780/CP-20腹膜内模型中，纳米型体在体内通过体内递送BMI-1 siRNA后卵巢肿瘤的生长和结节形成显着降低。直接靶向BMI-1 mRNA的siRNA或microRNA-15A/16对BMI-1的下调导致抑制增殖，克隆原性和敏化细胞对顺铂诱导的细胞凋亡。因此，我们假设BMI-1的下调将抑制卵巢肿瘤的生长，并在临床前小鼠模型中使肿瘤细胞对顺铂敏感。方法：我们将测试通过注射A-2780或CP-70（顺铂耐药）凝胶产生的原位腹膜内卵巢肿瘤小鼠模型的治疗功效。为了模拟siRNA/miR的siRPLATIN和/或纳米脂质体共轭物的晚期治疗，将在肿瘤接种后1周进行腹膜内治疗，持续4周。我们将确定（i）如果由siRNA敲除BMI-1，（ii）如果miR-15a/16敲除BMI-1会抑制肿瘤的生长并使细胞在体内敏感到顺铂；（iii）或者，将利用shRNA在A-2780和CP-70细胞中shRNA对BMI-1诱导的有条件敲低的条件敲低；（iv）通过蛋白质印迹分析，RT-PCR和免疫组织化学（IHC）验证DDR和凋亡途径的分子靶标，如AIM 1所示。拟议的研究将使用易于翻译的纳米脂质体递送方法验证BMI-1作为重要的新靶标，即使在化学耐药性卵巢癌中也有可能进行治疗。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Role of TGF-β signaling in uterine carcinosarcoma.

DOI：
10.18632/oncotarget.3711
发表时间：
2015-06-10
期刊：
Oncotarget
影响因子：
0
作者：
Dwivedi SK;McMeekin SD;Slaughter K;Bhattacharya R
通讯作者：
Bhattacharya R

MDR1 mediated chemoresistance: BMI1 and TIP60 in action.

DOI：
10.1016/j.bbagrm.2016.06.002
发表时间：
2016-08
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Banerjee Mustafi S;Chakraborty PK;Naz S;Dwivedi SK;Street M;Basak R;Yang D;Ding K;Mukherjee P;Bhattacharya R
通讯作者：
Bhattacharya R

Therapeutic evaluation of microRNA-15a and microRNA-16 in ovarian cancer.

DOI：
10.18632/oncotarget.7618
发表时间：
2016-03-22
期刊：
Oncotarget
影响因子：
0
作者：
Dwivedi SK;Mustafi SB;Mangala LS;Jiang D;Pradeep S;Rodriguez-Aguayo C;Ling H;Ivan C;Mukherjee P;Calin GA;Lopez-Berestein G;Sood AK;Bhattacharya R
通讯作者：
Bhattacharya R

Bmi-1: At the crossroads of physiological and pathological biology.