Combine mitochondrial gene therapy and synthetic lethal chemotherapy to treat triple-negative breast cancer

结合线粒体基因疗法和合成致死化疗治疗三阴性乳腺癌

• 批准号: 10642815
• 负责人: Xiaoguang Margaret Liu
• 金额: $ 56.16万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-10 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642815
• 关键词: 4T1; Achievement; Aftercare; Anthracycline; Apoptosis; Biodistribution; Bioluminescence; Biomanufacturing; Bioreactors; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer Treatment; Breast Cancer cell line; Bypass; CD276 gene; Cell Death; Cell physiology; Cells; Chemotherapy and/or radiation; Clinic; Clinical Trials; Cytoplasm; Cytosol; Cytotoxic Chemotherapy; DNA Damage; DNA Repair; DNA Repair Disorder; DNA Repair Inhibition; Dependovirus; Distant; Drug resistance; Elements; Engineering; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Evaluation; Flow Cytometry; Gene Expression; Genes; Heterogeneity; Histology; Histopathology; Human; Immune; Immunity; Immunocompetent; Immunohistochemistry; Impairment; Infiltration; Injections; Inner mitochondrial membrane; Investigation; Life; Literature; Luciferases; Macrophage; Malignant Neoplasms; Mediating; Membrane; Membrane Potentials; Metastatic/Recurrent; Microarray Analysis; Mitochondria; Modeling; Monoclonal Antibodies; Neoplasm Metastasis; Oncology; Organ; PARP inhibition; Pathology; Patients; Play; Poly(ADP-ribose) Polymerase Inhibitor; Polymerase; Pre-Clinical Model; Proteins; Proteomics; Quality of life; Recurrence; Research; Role; Shrimp; Signal Transduction; Signal Transduction Pathway; Specificity; Survival Rate; Technology; Testing; Tissue Microarray; Tissues; Toxic effect; Toxicology; Transcript; Translations; Transplantation; Treatment Efficacy; Treatment Protocols; Tumor Burden; Tumor Immunity; Tumor Volume; Western Blotting; Xenograft Model; Xenograft procedure; anti-cancer; brca gene; cancer cell; cancer drug resistance; cancer subtypes; chemotherapy; cytotoxic CD8 T cells; deep ocean; delivery vehicle; dosage; drug resistance development; exosome; gene therapy; immune function; immunoregulation; improved; in vivo; inhibitor; innovation; lapatinib; light gated; luciferin; nanoluciferase; novel strategies; overexpression; patient derived xenograft model; patient response; prevent; promoter; repaired; response; synergism; systemic toxicity; targeted delivery; targeted treatment; taxane; transduction efficiency; treatment strategy; triple-negative invasive breast carcinoma; tumor; tumor microenvironment; ultrasound

SUMMARY Triple-negative breast cancers (TNBCs) are highly aggressive and standard cytotoxic chemotherapies (e.g. anthracycline-taxane) are the main treatment strategies in clinics. Our previous research and literature demonstrated that poly (ADP-Ribose) polymerase inhibitors (PARPi) as single agents or in combination with epidermal growth factor receptor inhibitor (EGFRi) induced a contextual synthetic lethality and reduced TNBC metastsis by inhibiting the repair of DNA damage. Our clinical trial showed that veliparib (PARPi)/lapatinib (EGFRi) achieved a 24% response rate in TNBC patients with wildtype BRCA1/2. Despite these achievements, TNBC cells often develop drug resistance to chemotherapies, have low patient response rate, and regrow after primary treatment. Novel strategies that can effectively treat TNBCs are urgently needed. Mitochondria are the powerhouse of cells and play a pivotal role in regulating cell functions, rendering them a promising oncological target. Destroying mitochondrial function, such as directly depolarizing the inner mitochondrial membrane (IMM) potential via synthesized heterologous genes, can bypass the repair of signaling transduction pathways, trigger a point-of-no-return cancer cell death, and subsequently prevent the development of drug resistance. We recently developed a mitochondrial luminoptogenetics (mLumiOpto) technology by synthesizing the heterologous light-gated channelrhodopsin in IMM and an engineered luciferase in cytoplasm, which induces IMM depolarization through opening mitochondrial channelrhodopsin with luciferase-luciferin emitted endogenous blue bioluminescence. Preliminary studies showed that mLumiOpto effectively depolarized mitochondria in TNBC cell lines representing multiple subtypes, resulted in persistent DNA damage, and significantly reduced tumor burden in three TNBC xenograft models. Applying our dual-targeted delivery vehicle, i.e. EGFR/CD276 monoclonal antibodies tagged exosome-associated adeno-associated virus (mAb-Exo-AAV), and cancer-specific promoter (cfos) in mLumiOpto achieved high TNBC specificity, functional expression, and minimal undesirable systemic toxicity. The objective of this project is to harness the combination of targeted mLumiOpto that is delivered with mAb-Exo-AAV and PARPi to eliminate TNBC cells in vivo. The hypothesis is that the combined mLumiOpto/PARPi integrates multiple anti-cancer mechanisms, i.e., IMM depolarization, DNA damage and inhibition of repair, and tumoral immunity. Specifically, large-scale dual-targeted mLumiOpto will be generated and characterized; treatment dosage and strategy will be optimized; and anti-cancer efficacy will be evaluated in TNBC primary xenograft model and distant metastatic model (Aim 1). Furthermore, the synergistic effects of mLumiOpto/PARPi will be assessed and the underlying mechanisms will be investigated in immunocompetent models (Aim 2). Finally, the metastasis reduction and heterogeneous TNBC treatment efficacy will be fully tested in metastatic and patient-derived xenograft (PDX) models, and toxicology will also be investigated (Aim 3). Successful completion of this project will provide a new strategy to treat TNBCs.

概括 三阴性乳腺癌（TNBC）是高度侵略性和标准的细胞毒性化学疗法（例如， Anthracycline-Taxane）是诊所的主要治疗策略。我们以前的研究和文学 证明聚（ADP-核糖）聚合酶抑制剂（PARPI）作为单一药物或与 表皮生长因子受体抑制剂（EGFRI）诱导了上下文合成的致死性，TNBC降低 转移通过抑制DNA损伤的修复。我们的临床试验表明，维利帕里（PARPI）/lapatinib （EGFRI）在WildType BRCA1/2患者的TNBC患者中达到了24％的缓解率。尽管取得了这些成就， TNBC细胞通常会产生对化学疗法的耐药性，患者的反应率较低，并且在 初级治疗。迫切需要有效治疗TNBC的新型策略。线粒体是 细胞的动力室，并在调节细胞功能中起关键作用，使它们成为有希望的肿瘤学 目标。破坏线粒体功能，例如直接去极化线粒体膜（IMM） 通过合成的异源基因的电势可以绕过信号转导途径的修复，触发 无返回的癌细胞死亡，随后防止耐药性的发展。我们 最近开发了一种通过合成的线粒体发光剂（mlumiopto）技术 IMM中的异源光门控通道旋转和细胞质中的工程荧光素酶，可诱导诱导 免疫去极化，通过发出的线粒体通道旋转蛋白发出荧光素酶 - 卢西蛋白发出 内源性蓝色生物发光。初步研究表明，mlumiopto有效去极化 TNBC细胞系中表示多种亚型的线粒体，导致持续的DNA损伤，并 在三种TNBC异种移植模型中，肿瘤负担大大减轻了。应用我们的双目标送货工具， 即EGFR/CD276单克隆抗体标记与外泌体相关的腺相关病毒（MAB-EXO-AAV）， Mlumiopto中的癌症特异性启动子（CFO）可实现高TNBC特异性，功能表达和 最小的不良系统毒性。该项目的目的是利用目标的组合 用mab-exo-aav和parpi传递的mlumiopto在体内消除TNBC细胞。该假设是 合并的mlumiopto/parpi整合了多种抗癌机制，即免疫去极化，DNA 损害和抑制修复和肿瘤免疫。具体而言，大规模双重目标的mlumiopto将是 生成和表征；将优化治疗剂量和策略；和抗癌功效将是 在TNBC主要异种移植模型和远处转移模型中进行了评估（AIM 1）。此外，协同作用 将评估mlumiopto/parpi的影响，并将研究基本机制 免疫能力模型（AIM 2）。最后，减少转移和异质TNBC治疗 功效将在转移性和患者衍生异种移植（PDX）模型中进行全面测试，毒理学也将是 调查（AIM 3）。该项目的成功完成将为治疗TNBC提供新的策略。