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.

摘要 三阴性乳腺癌(TNBCs)是一种高度侵袭性和标准的细胞毒性化疗方法(例如： 是临床上的主要治疗策略。我们以前的研究和文献 证明了聚(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)诱导背景合成致死性并降低TNBC 通过抑制DNA损伤的修复而发生转移。我们的临床试验显示维拉帕利(PARPI)/拉帕替尼 (EGFRi)在患有野生型BRCA1/2的TNBC患者中实现了24%的应答率。尽管取得了这些成就， TNBC细胞通常对化疗产生抗药性，患者应答率低，并在 初级治疗。迫切需要能够有效治疗TNBCs的新策略。线粒体是 细胞的动力库，在调节细胞功能方面发挥着关键作用，使它们成为一种有前途的肿瘤学 目标。破坏线粒体功能，如直接使线粒体内膜去极化 潜在的通过合成异源基因，可以绕过信号转导通路的修复，触发 一种点不归路的癌细胞死亡，从而阻止耐药性的发展。我们 最近开发了一种线粒体光遗传学(MLumiOpto)技术，通过合成 IMM中异源光门通道视紫红质和细胞质中的工程荧光素酶诱导 开放线粒体通道的IMM去极化视紫红质与荧光素酶-荧光素释放 内源蓝色生物发光。初步研究表明，mLumiOpto有效地去极化 代表多种亚型的TNBC细胞系中的线粒体，导致持续的DNA损伤，以及 在三种TNBC异种移植模型中显著降低了肿瘤负担。应用我们的双目标送货工具， 即标记外切体相关腺相关病毒的EGFR/CD276单抗(mAb-Exo-AAV)， 和mLumiOpto中的肿瘤特异性启动子(CFO)获得了高TNBC特异性、功能性表达和 最小的不良全身毒性。该项目的目标是利用有针对性的组合 MLumiOpto与mAb-Exo-AAV和PARPI一起传递，在体内消除TNBC细胞。假设是 MLumiOpto/PARPI结合了多种抗癌机制，即IMM去极化，DNA 损伤和修复抑制，以及肿瘤免疫。具体地说，大规模双靶mLumiOpto将 产生和表征；治疗剂量和策略将优化；抗癌效果将 在TNBC原发异种移植模型和远处转移模型中进行评估(目标1)。此外，协同效应 将评估mLumiOpto/PARPI的效果，并将在#年调查其潜在机制 免疫活性模型(目标2)。最后，减少转移和异种TNBC治疗 疗效将在转移性和患者来源的异种移植(PDX)模型中得到充分测试，毒理学也将得到 调查(目标3)。该项目的成功完成将为治疗TNBCs提供一种新的战略。