Targeted Exosome-Associated AAV-Mediated Gene Therapy to Eliminate Metastatic Neuroendocrine Cancers

靶向外泌体相关 AAV 介导的基因治疗可消除转移性神经内分泌癌

基本信息

批准号：
9907789
负责人：
Joseph N Garner
金额：
$ 24.31万
依托单位：
TECHNOLOGY COMMERCIALIZATION PARTNERS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9907789
关键词：
Affinity Animal Model Animals Apoptosis Binding Biodistribution Biological Markers Biological Response Modifier Therapy Bioluminescence Biomanufacturing Bioreactors Bypass Cancer Burden Cancer Patient Cancer cell line Carcinoid Tumor Cell Death Cells Cessation of life Clinic Clinical Trials Cloning Coupled Cytosol Data Dependovirus Diagnosis Diarrhea Drug Kinetics Electron Microscope Endocrine system Energy Metabolism Engineering Enzymes Excision Flow Cytometry Flushing Future Gene Delivery Gene Proteins Genes Goals Heart failure Hormone secretion Hormones IgG1 Impairment In Vitro Inner mitochondrial membrane Islet Cell Tumor Islets of Langerhans Legal patent Light Liver Luciferases Lung Malignant - descriptor Malignant Carcinoid Syndrome Malignant Neoplasms Maximum Tolerated Dose Mediating Membrane Potentials Metastatic Neoplasm to the Liver Mitochondria Modeling Monoclonal Antibodies Morphology Mus Names Neoplasm Metastasis Neoplasms Nervous system structure Neuroendocrine Tumors Neurosecretory Systems Normal Cell Octreotide Operative Surgical Procedures Pancreas Patients Peptide Receptor Pharmaceutical Preparations Play Property Proteins Protons Quality of life Radiation therapy Resistance Rhodopsin Role SDZ RAD SSTR2 gene Safety Scheme Signal Transduction Source Surface Survival Rate System Techniques Technology Therapeutic Therapeutic Effect Time Translations Treatment Efficacy Western Blotting Xenograft procedure anti-cancer base cancer cell cancer therapy chemotherapy clinical efficacy confocal imaging exosome expression vector gene therapy imaging system improved in vivo in vivo imaging system innovation light gated luciferin medullary thyroid carcinoma mitochondrial membrane mouse model neuroendocrine cancer novel optogenetics pre-clinical promoter protein expression response side effect somatostatin receptor 2 subcutaneous targeted treatment therapeutic gene transmission process tumor tumor growth

项目摘要

ABSTRACT Neuroendocrine (NE) malignancies are hormone secreting neoplasms that arise from endocrine and nervous system. Multiple NE tumors (NETs) have been diagnosed, such as pancreatic neuroendocrine cancers, medullary thyroid cancers, and pulmonary neuroendocrine carcinoids. Most NE cancer patients are metastatic at the time of initial diagnosis which makes the complete resections via surgery impossible. The current chemotherapies, including Octreotide, Sunitinib, Everolimus and peptide receptor, have marginal curative benefits and severe side effects. Thus, an effective targeted therapy is critical for patients with metastatic NE cancers. We have recently developed a novel technique, named “mitochondrial chemo-optogenetics”, by expressing a heterologous light-gated channelrhodopsin protein in the IMM of cancer cells, and depolarizing IMM potentials and inducing cell death by using luciferase-luciferin bioluminescence as the endogenous light source. Our preliminary data showed that this new mitochondrial gene therapy caused substantial NE cancer cell death in vitro and stopped NE tumor growth and even reduced tumor size in a subcutaneous NE cancer xenograft mouse model. Additionally, we have built an innovative NE cancer-targeted gene delivery platform by tagging our new anti-somatostatin receptor 2 (SSTR2) monoclonal antibody (mAb) to the surface of exosome. However, a targeted gene therapy, such as mAb-Exo-AAV carrying our mitochondrial chemo-optogenetics therapeutic gene, is urgently needed to achieve substrate-induced mitochondrial depolarization and selective elimination of cancer cells in vivo. Moreover, the therapeutic efficacy of the gene therapy in metastatic a model is essential because most diagnosed NE cancer patients are metastatic. The specific objective of this application is to develop, produce and evaluate an innovative NE cancer-targeted mitochondrial gene therapy to selectively destroy and eliminate NETs in vivo. The following two specific aims over a 12-month period are propose. Aim 1: To develop, produce and characterize the NE-cancer targeted mitochondrial gene therapy. A high-quality anti-SSTR2 mAb-Exo-AAV will be constructed by cloning a cancer promoter (cfos) and the fused blue light- producing luciferase and light-gated rhodopsin gene, i.e. cfos-NLuc-2A-ABCB-CoChR (~3.3 kb), into the engineered pAAV-MCS promoterless expression vector, and produced using our stirred-tank bioreactor-based exosome-AAV biomanufacturing platform and surface tagging technology. The anti-SSTR2 mAb-Exo-AAV will then be evaluated for its cancer specific targeting and in vitro anti-cancer efficacy. Aim 2: To evaluate the therapeutic values of the mitochondrial gene therapy using preclinical NET metastatic animal model. Most NE cancer patients are initially diagnosed with metastases and have already developed carcinoid syndrome. Therefore we will evaluate the maximal tolerated dose (MTD), pharmacokinetics (PK), anti- NET efficacy, and liver metastases reduction of the developed gene therapy using metastatic model.

抽象的神经内分泌（NE）恶性肿瘤是马龙分泌肿瘤，由内分泌和紧张产生系统。已经诊断出多个NE肿瘤（NET），例如胰腺神经内分泌癌，甲状腺癌和肺神经内分泌类癌。大多数NE癌症患者是转移性的在初始诊断时，通过手术使完整的切除不可能。电流化学疗法，包括奥曲肽，舒尼替尼，依西洛木和肽受体，具有边缘治疗好处和严重的副作用。这是有效的靶向疗法对于转移性NE的患者至关重要癌症。我们最近开发了一种新颖的技术，称为“线粒体化学渗透遗传学”，通过表达癌细胞INM中异源光门控通道旋丁蛋白蛋白，并使INM电位分裂并通过使用荧光素酶卢西蛋白生物发光作为内源光源来诱导细胞死亡。我们的初步数据表明，这种新的线粒体基因疗法在体外并停止NE肿瘤生长，甚至减少皮下NE癌症小鼠的肿瘤大小模型。此外，我们通过标记我们的新的，建立了一个创新的NE癌症基因输送平台抗阳性蛋白蛋白受体2（SSTR2）单克隆抗体（MAB）与外泌体表面。但是，有针对性的基因疗法，例如携带线粒体化学遗传学的mab-exo-aaV 迫切需要治疗基因来实现底物诱导的线粒体沉积和选择性消除体内癌细胞。此外，基因治疗在转移性A模型中的治疗效率这是必不可少的，因为大多数被诊断的NE癌症患者都是转移性的。该应用程序的具体目标是开发，生产和评估以癌为癌的线粒体基因治疗的创新性NE 在体内销毁和消除网。在12个月内，以下两个特定目标是提议。目标1：开发，生产和表征NE-CANCER靶向线粒体基因治疗。高质量通过克隆癌症启动子（CFO）和融合的蓝光 - 产生荧光素酶和光门控的红宝石基因，即CFOS-NLUC-2A-ABCB-COCHR（〜3.3 Kb）设计了PAAV-MCS启动子表达矢量，并使用我们的升降机生物反应器生产外部AAV生物制造平台和表面标记技术。抗SSTR2 mab-exo-aav将然后对其癌症特异性靶向和体外抗癌效率进行评估。目标2：使用临床前净转移性评估线粒体基因治疗的治疗值动物模型。大多数NE癌症患者最初被诊断为转移，并且已经开发类癌综合征。因此，我们将评估最大耐受剂量（MTD），药代动力学（PK），抗 - 净效率和使用转移模型减少开发基因治疗的肝转移。