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Normalizing aberrant metabolism in ovarian cancer by a unique drug delivery system

通过独特的药物输送系统使卵巢癌的异常代谢正常化

基本信息

批准号：
10545752
负责人：
Resham Bhattacharya
金额：
$ 37.61万
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：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10545752
关键词：
Animal Model Apoptosis Biodistribution Biological Assay Cancer cell line Cell Culture Techniques Chemicals Cisplatin Clathrin Clinic Clinical Drug Delivery Systems Drug Kinetics Drug Sensitization Drug resistance Endocytosis Enzymes Exhibits Formulation Future Gene Silencing Glycolysis Gold Growth Human Immune system Immunofluorescence Immunologic In Vitro Inner mitochondrial membrane Invaded Lipofectamine Liposomes Location Lysosomes Malignant neoplasm of ovary Mediating Membrane Proteins Metabolic Metabolism Mitochondria Modeling Mus Nanoconjugate Normal Cell Outcome Oxidative Phosphorylation Pathway interactions Patients Phenocopy Phenotype Prognosis RNA delivery Reagent Reporting Resistance Rod Scheme Serous Shapes Small Interfering RNA System Testing Therapeutic Tissue Microarray Tissues Toxic effect Transfection Translating Translations Treatment Efficacy Western Blotting Xenograft procedure cancer cell clinical translation genetic approach in vivo inhibitor liposomal formulation migration mouse model nanoGold nanocomposite nanoformulation nanoliposome nanotoxicity new therapeutic target overexpression patient derived xenograft model pharmacologic pyruvate dehydrogenase targeted treatment therapeutic RNA therapeutic gene translatable strategy tumor tumor growth tumor microenvironment uptake

项目摘要

Gene silencing using small interfering RNA (siRNA) is a viable therapeutic approach but, limited in translation due to lack of effective delivery systems. Developing effective and non-toxic delivery system will translate siRNA- based therapeutics to clinics. Here, using in vitro cell culture and in vivo animal models, we propose to develop a new type of siRNA delivery system for effective gene silencing and therapeutic applications. We recently reported that MICU1, a mitochondrial inner membrane protein, functions as a metabolic switch that promotes glycolysis and therapy resistance in ovarian cancer. Unfortunately, lack of pharmacological inhibitors and effective strategies to silence MICU1 in vivo posit a significant challenge against future clinical translation of MICU1-targeted therapy. Therefore, MICU1 could serve as a new therapeutic target to validate silencing and therapeutic efficacy of our new siRNA delivery platform and provides opportunity to normalize aberrant metabolism responsible for therapy resistance. Hence, we plan to develop a gold nanoparticle (AuNP)- based liposomal formulation (AuroLiposome) for siRNA delivery to effectively silence MICU1 in vivo. To effectively silence MICU1 in vivo we have developed DOPC-DOTAP based conventional nanoliposomal siRNA delivery platforms (MICU1 siRNA-cLPs). Interestingly, AuNP (20 nm size)-doped formulation (MICU1 siRNA-AuroLPs) exhibited enhanced efficacy in silencing MICU1, requiring 3-4-fold lower siRNA concentrations than MICU1 siRNA-cLPs or commercially available transfection reagents such as Hiperfect, RNAiMax and Lipofectamine 3000. Enhanced silencing was reflected in clonal growth assays; MICU1 siRNA-AuroLPs inhibited clonal growth of HGSOCs more efficiently (~90%) than MICU1 siRNA-cLPs (~50%) or Hiperfect (~30%). Importantly MICU1 siRNA-AuroLPs inhibited tumor growth more effectively (~75%) compared to MICU1 siRNA-cLPs (~35 %). Importantly, using chemical inhibitors we showed that incorporation of AuNP switched intracellular uptake pathway of MICU1 siRNA-cLPs from a combination of clathrin and caveolar mediated endocytosis to mostly caveolar uptake pathway. Hence, we hypothesize that incorporation of AuNP in nanoliposomal formulation triggers caveolar uptake of AuroLiposome (AuroLPs) resulting in reduced degradation of siRNA-AuroLPs in lysosome and thus enhancing silencing efficacy. We will use specific aims below to test the hypothesis and accomplish overall objectives; Aim1: Determining mechanisms of enhanced gene silencing efficacy due to gold doping. Aim 2: Determining pharmacokinetics, biodistribution and toxicity of the optimized nanoformulation. Aim 3: Determining therapeutic efficacy in patent derived xenografts (Pdx) and syngeneic mouse model. Successful completion of the project will provide a generalized siRNA delivery approach for any in vitro and in vivo gene silencing applications and a potential translatable strategy to normalize aberrant metabolism to overcome therapy resistance against high grade serous ovarian cancer.

使用小干扰RNA （siRNA）进行基因沉默是一种可行的治疗方法，但在翻译方面受到限制